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Expression analysis and functional characterization of the dro1/Cl2 gene during zebrafish somitogenesis

Conference Paper · July 2009 DOI: 10.13140/2.1.2295.8407

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Program & Abstracts *U
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SFTFBSDIFST 6th European Zebrafish Genetics and Development Meeting Rome 15th - 19th July 2009

Program & Abstracts Intro 2 Meeting information. Registration. Organizing Committee. Congress map. Program ataglance. Aknowledgements. Information for Authors. Exhibitors andExhibitionarea. Sponsors. ABSTRACTS. Scientific program. Session XII: Session XI: Session X: Session IX: Session VIII: Session VII: Session VI: Session V: Session IV: Session III: Session II: Session I: TALKS ...... Neurobiology II:sensoryorgansandregeneration. Neurobiology I:patterningandbehaviour. Hematopoiesis andimmune system. Cardiovascular system. Disease models. Cancer. Genomic workshop. Emerging technologies. Organogenesis. Gene regulation. Signaling. Early development, gastrulationandsegmentation...... Contents Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. 104 10 12 11 14 17 74 98 92 86 80 67 61 55 49 43 37 35 5 4 6 9 8 Authors index . Gene regulation. Segmentation. Cell movement. Live imaging. Emerging technologies. Bioinformatics andsystemsbiology. Genomics. MicroRNA andnon-codingRNAs. Regeneration. Cancer. Disease models. Hematopoiesis andimmunology. Cardiovascular system. Behaviour. Neurobiology: sensoryorgans. Neurobiology: patterning. Organogenesis. Signaling. Early development andgastrulation. Gern cellsandmaternaldeterminants. Husbandry workshop. POSTER ...... Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. Pag. 521 496 487 479 223 474 458 457 441 437 425 405 351 325 297 293 268 188 164 143 149 147 3

Intro Intro 4 IFOM, The Firc InstituteofMolecularOncology, Milan Temasek LifeSciencesLaboratory, Singapore University ofPadua, Padua, Italy The MinistryofForeign Affairs University of Turin, Turin, Italy Organizing committee Organizing Acknowledgements Under theauspicesof Federica Pezzimenti Francesco Argenton Gianluca Deflorian Cristina Santoriello Massimo Santoro Jachen Solinger Karuna Sampath Marina Mione Viviana Anelli Carlo Titone Payments accepted“onsite”arecashandcreditcard. PAYMENT CONDITIONS and Secretariat the at so do to able Reception DeskattheCongress be Venue. will registered not have who Participants REGISTRATION • CoffeebreaksandLunches thatwillbeheldattheCongress Venue. • Welcome receptionon Wednesday 15th,2009 • Badgeandcongress Welcome kit • Program andabstracts book • Entrance totheexhibitionarea • Attendance toallscientificsessions The registration feeincludes: Industry University Faculty Students /postdocs REGISTRATION FEES(VAT 20%INCLUDED) Registration € 600,00 € 490,00 € 420,00 Before 30/03/2009 € 750,00 € 630,00 € 550,00 After 30/03/2009 5

Intro Intro 6 of properties private to damages to maketheir own arrangements iftheyconsider itnecessary. or loss accidents, participants and exhibitors can not be personal accepted. Participants and exhibitors are advised for Responsibility Insurance The officiallanguageoftheMeetingis English. Official Language The cloackroom islocatedintheLounge. Cloakcroom andLuggage An InternetPoint isavailable intheFuniRoom. Internet Point the Meeting. Certificate of Attendance will be requested at the Reception Desk and sent by mail after Certificate of Attendance rooms. You shouldalsowearyour badgeintheexhibitionarea. meeting the to admitted be will badge name their wearing are who gathering. participants Only social or session scientific any attending when badge name your wear to All participants will receive a personal badge upon registration. YouBadges are kindly requested the follow will activities and Cultura della scientific sessions’timetable. Salone the in placed is area Exhibition Exhibition Area - Sunday - Saturday - Friday - Thursday - Wednesday Opening hours: within thefollowing times: The Organizing Secretariat will be available at the Congress Venue during the meeting e-mail: [email protected] -www.adriacongrex.it Via Sassonia, 30 - 47900 Rimini (RN) - Italy - Tel. +39.0541.305863 - Fax +39.0541.305842 Adria CongrexandMeetitalyGroup Organizing Secretariat Piazzale John Kennedy, 1-00144Roma -Italy Palazzo deiCongressi Congress Venue July 19th,2009 July 18th,2009 July 17th,2009 July 16th,2009 July 15th,2009 9.00-13.00 9.00-18.00 9.00-19.30 9.00-18.00 13.00-20.00 Meeting Information t il e osbe o eitr o te oil inr lo uig h Meig t the at Meeting Payments acceptedarecashandcreditcard. the during also Dinner Social the Secretary Desk. for register to possible be will It Rates include:Buffet,drinksandLive Music. University Faculty andIndustry Student/Post Docs Rates: Venue: Centrale Ristotheatre Time: 20.00 Date: 18thJuly 2009 SOCIAL DINNER Price: freeforallregistereddelegates. Venue: Palazzo deiCongressi-Exhibitionarea Time: 18.00 Date: Wednesday 15thJuly 2009 WELCOME RECEPTION SOCIAL PROGRAM Electricity suppliedis220V-50Hz. the and Electricity supply Committee Organizing the reason, technical Organizing Secretariatreserve and/or therighttomakechanges intheMeetingprogram. scientific any For Changes inProgram Public Parking isavailable inthe area. Parking attheCongress Venue the in served be will fee) registration Exhibition Area intheSalonedellaCultura. the in (included lunches and breaks Coffee Coffee breaksandLunches € 70,00 Vat included € 60,00 Vat included 7

Intro Intro 8 DELL AMB DELL AMB SAL DELLE SCIENZE: A SAL UDIT A R A FUNI: UL UL A LET A PIT DELLA LETTERATURA:DELLA AMBULACRO PITTURA: DELLA AMBULACRO ARTE: SALA DELLE SCIENZE: AUDITORIUM FUNI: SALA ORIUM A A CENTER SAL TE: FUNI SLIDE CR CR Bar TUR TER O O AMBULACRO DELLA PITTURA A CENTER SALA FUNI SLIDE Bar A: A TUR AMBULACRO DELLA PITTURA C CONGRESS MAP ONGRESS MAP A: U.S. U.S.

U.S. U.S. U.S. U.S. POSTER AREA POSTER AREA T SANGER CENTER ZIR U.S. U.S. SAL MAIN ENTRANCE ALK SESSIONSANDKE POSTER AREA POSTER AREA ZIRC, ZHAand UCLHUSBANDRY WORKSHOP TALK SESSIONANDKEYNOTE LECTURE SANGER CENTER WORKSHOP -INTERNETPOINT SALONE CULTURA DELLA MAIN ENTRANCE C, WEL EXHIBITION AREA ONE DELL WELCOME RECEPTION EXHIBITION AREA ZHAandUCLHUSBANDR C A C CATERING AREA A AUDITORIUM Atrio Kennedy Piazzale REGISTRA UDIT ENTRANCE OME RECEPTION PLENAR TERING AREA Atrio Kennedy Piazzale REGISTRATION SESSION ENTRANCE SCIENZE PLENARY SESSION SCIENZE DELLE DELLE DESK

DESK

ORIUM

A CUL Arte Arte TION Y TUR WORKSHOP -INTERNETPOINT U.S. U.S. A U.S. U.S.

YNO U.S. U.S.

U.S. U.S. TE LECTURES SALA ARTE Y

WORKSHOP AMBULACRO DELLA LETTERATURA SAL AR Congress Map TE

AMBULACRO DELLA LETTERATURA A Sponsors Large ParticleFlowCytometry 9

Intro oeGntc N. 9 N. 19 Müller &Pfleger GmbH &Co.KG N. 5 Genetics Mole N. 3 Leica Microsystems Intavis Instruments Bioanalytical AG N.N. 14 15 Gene Tools N. 1 AG Eppendorf N. 6–7 N.17 Aquatic Habitats Aquarien-Bau Schwarz Aquaneering Company

Intro oeGntc N. 9 N. 19 Müller &PflegerGmbH &Co.KG N. 5 Genetics Mole N. 3 Leica Microsystems Intavis Instruments Bioanalytical AG N.N. 14 15 Gene Tools N. 1 AG Eppendorf N. 6–7 N.17 Aquatic Habitats Aquarien-Bau Schwarz Aquaneering Company 10 OPENED EDGE OPENED EDGE 17 19 17 19 EXHIBITION AREA 15 EXHIBITION AREA 15 Booth Booth 27 14 27 14 REGISTRATION REGISTRATION 21 21 1 1 CATERING CATERING 26 13 Company iwon N. 13 N. 25 N. 24 N. 12 N. 21–2223 ZebrafishZIRC International Resource Center ZFIN Zebrafish Information Network N. 26 ViewPoint Union Biometrica N. 11 Tecniplast N. 27 InstituteSanger Noldus Information Technology NIBB -National Institute for Biology Basic 26 13 AREA Company iwon N. 13 N. 25 N. 24 N. 12 N. 21–22 – 23 ZebrafishZIRC International Resource Center ZFIN Zebrafish Information Network N. 26 ViewPoint Union Biometrica N. 11 Tecniplast N. 27 InstituteSanger Noldus Information Technology NIBB -National Institute for Biology Basic DESK AREA DESK 22 3 22 3 25 12 25 12 23 5 23 5 24 11 24 11 9 7 6 Exhibition Area 9 7 6 Exhibitors Booth Booth check that everything works at the Slide Center at least 2 hours before your session yourstarts. before hours 2 least at Center Slide the at works everything that check please laptop, your using are you If starts. session your before hours 2 least at Center Slide the at ROM – cd the or drive during pen a availableon presentator be your deliver Please will conference. center slide A program. video EO the through accepts PowerPointthat format a to converted be will they work not do movies If embedded. be should they presentation the during used are movies resolutionWhen pixels 768 x 1024 a in equivalent) (or PowerPointfiles as prepared be should presentations All conference All discussion. minutes 5 rooms fororal presentationshave laptopcomputersandvideoprojectors. plus long minutes 10 are presentations Oral Oral presentations even numbers). 18th: July numbers, odd 17th: (July program the in indicated as session poster the during poster their at present be to requested are authors Poster Desk. Secretariat Assistance andmaterialformountingposterswillbeavailable fromset-uptime,atthe and destroyed by theOrganizingSecretariat. by removed be to has It Sunday, panel. July numbered 19th, 2009 before 11.00. corresponding If posters are not removed, the they will be in removed 13.00 at 15th July from mounted be can poster high. The cm 100 x wide cm. 70 are dimensions board Poster 2009. 19th, July to 15th July from PosterArea the in display on be will Poster Poster Sessions Information for Authors 11

Intro Intro 12 WE WOULD LIKE TO THANK ALL ABSTRACT REVIEWERS S. Schulte-Merker, The Netherlands C.P. Heisenberg,Germany M.V. Flores,NewZealand G. Weidinger, Germany H. Lopez-Schier, Spain R. Dosch, Switzerland D. Gilmour, Germany G. Lieschke, Australia M. Schartl, Germany R. Koester, Germany H. Okamoto,Japan G. Lefkowitz, Israel J. Topczewski, USA A. Oates,Germany J. Amatruda, USA M. Halpern,USA A. Giraldez, USA D. Meyer, Austria T. Schilling, USA T. Whitfield, UK D. Stemple, UK E. Knapik,USA C. Houart,UK F. Mueller, UK N. Trede, USA C. Becker, UK M. Tada, UK E. Ober, UK Acknowledgements

Intro 14 16:30 -18:00 16:00 -16:30 14:30 -16:00 12:30 -14:00 11:00 -12:30 10:30 -11:00 18:00 -20:00 17:00 -18:00 14:00 -16:00 13:30 -16:30 12:45 -13:30 09:15 -10:45 11:00 -12:45 10:45 -11:00 09:00 -10:30 SESSION I-Earlydevelopment, gastrulationandsegmentation AUDITORIUM DELLE KEYNOTE LECTUREI Stefano Piccolo in development TGFb signaling and cancer SCIENZE COFFEE BREAK AT SALONEDELLACULTURA COFFEE BREAK AT SALONEDELLACULTURA WELCOME RECEPTION AT SALONEDELLACULTURA LUNCH AT SALONEDELLACULTURA SESSION III-Generegulation SESSION IV-Organogenesis AUDITORIUM DELLESCIENZE COFFEE BREAK AT SALAFUNI SESSION II-Signaling WEDNESDAY, JULY 15th,2009 THURSDAY JULY 16th,2009 SANGER INSTITUTE SANGER INSTITUTE SANGER INSTITUTE LUNCH BREAK WORKSHOP WORKSHOP WORKSHOP SALA FUNI UCL HUSBANDRY Program ataGlance ZIRC, ZHA, AND WORKSHOP SALA ARTE PITTURA E AMBULACRO POSTERS DISPLAYED DELLA LETTERATURA AMBULACRO DELLA POSTERS DISPLAYED AMBULACRO DELLA DELLA PITTURAE LETTERATURA AMBULACRO 16:30 -18:00 16:00 -16:30 14:30 -16:00 12:30 -14:00 11:00 -12:30 10:30 -11:00 09:00 -10:30 18:00 -20:00 17:00 -18:00 14:00 -16:00 13:30 -16:30 12:45 -13:30 11:00 -12:45 10:45 -11:00 09:15 -10:45 SESSION I-Earlydevelopment, gastrulationandsegmentation AUDITORIUM DELLE KEYNOTE LECTUREI Stefano Piccolo in development TGFb signaling and cancer SCIENZE COFFEE BREAK AT SALONEDELLACULTURA COFFEE BREAK AT SALONEDELLACULTURA WELCOME RECEPTION AT SALONEDELLACULTURA LUNCH AT SALONEDELLACULTURA SESSION III-Generegulation SESSION IV-Organogenesis AUDITORIUM DELLESCIENZE COFFEE BREAK AT SALAFUNI SESSION II-Signaling WEDNESDAY, JULY 15th,2009 THURSDAY JULY 16th,2009 SANGER INSTITUTE SANGER INSTITUTE SANGER INSTITUTE LUNCH BREAK WORKSHOP WORKSHOP WORKSHOP SALA FUNI UCL HUSBANDRY ZIRC, ZHA, AND WORKSHOP SALA ARTE PITTURA E AMBULACRO POSTERS DISPLAYED DELLA LETTERATURA AMBULACRO DELLA POSTERS DISPLAYED AMBULACRO DELLA DELLA PITTURAE LETTERATURA AMBULACRO Program ataGlance 18:00 -19:30 17:30 -18:00 16:00 -17:30 14:30 -16:00 12:30 -14:00 11:00 -12:30 20:00 -02.00 17:30 -18:00 16:00 -17:30 14:30 -16:00 12:30 -14:00 11:00 -12:30 10:30 -11:00 09:00 -10:30 09:30 -11:00 11:30 -13:00 11:00 -11:30 10:30 -11:00 09:00 -10:30 13:00 SESSION XII-NeurobiologyII: sensoryorgansandregeneration CONCLUDING REMARKS AND ENDOF THE MEETING SESSION XI-NeurobiologyI:patterningandbehaviour Enhancing mammalianregeneration -NadiaRosenthal SESSION X-Hematopoiesisamdimmunesystem COFFEE BREAK AT SALONEDELLACULTURA COFFEE BREAK AT SALONEDELLACULTURA COFFEE BREAK AT SALONE DELLACULTURA COFFEE BREAK AT SALONE DELLACULTURA COMMUNITY MEETING-MarinaMione LUNCH AT SALONEDELLACULTURA SESSION IX-Cardiovascular system SESSION V -Emergingtechnologies LUNCH AT SALONEDELLA CULTURA SESSION VI -Genomicworkshop AUDITORIUM DELLESCIENZE AUDITORIUM DELLESCIENZE AUDITORIUM DELLESCIENZE SESSION VIII -Diseasemodels KEYNOTE LECTUREII SESSION VII -Cancer SATURDAY JULY 18th,2009 SOCIAL DINNER SUNDAY JULY 19th,2009 FRIDAY JULY 17th,2009 AMBULACRO DELLA AMBULACRO DELLA AMBULACRO DELLA POSTER DISPLAYED POSTER DISPLAYED POSTER DISPLAYED POSTER DISPLAYED POSTER DISPLAYED DELLA PITTURAE DELLA PITTURAE DELLA PITTURAE EVEN NUMBERS ODD NUMBERS PRESENTATION PRESENTATION LETTERATURA LETTERATURA LETTERATURA AMBULACRO AMBULACRO AMBULACRO POSTERS POSTERS 15

Intro

13:00 -17:00 REGISTRATION DESK– ATRIO KENNEDY 15:45 -16:30 14:30 -15:45 13:30 -14:30 12:45 -13:30 11:00 -12:45 10:45 -11:00 09:30 -10:45 09:15 -09:30 09:15 -16:30 SALA FUNI Wednesday July 15 Scientific Program Participants’ Registration Working withZFIN Comparative Sequence Analysis Exploring proteinfunctionanddisease Lunch Break Genome Browsing Coffee Break Databases anddenovo analysisofsequence Introduction Only forpre-registeredparticipants Working withZebrafish GenomeResources Sanger Institute Workshop th , 2009 17

July 15th July 15th 18 14:00 -16:00 SALA ARTE

75 74 73 Chairs: in theUSandEU Raising Fish, NurseryOperations, and Animal Useregulations ZIRC, ZHA,andUCLHusbandry Workshop

Zoltan M. Varga, Eugene,USA US LegislationforFish HusbandryandShipping UCL Fish Facility, London,UK Carole Wilson EU LegislationforFish HusbandryandShipping Discussion Carrie Barton,Eugene,USA The ZIRCNurseryandGrow-Out Section Jenna LeaHakkesteeg,London,UK Zebrafish Larval RearingatUCL Federica Pezzimenti, Milan, Italy Fish BreedinginMilan Carole Wilson, London,UK Zoltan M. Varga, Eugene,USA Wednesday July 15 Scientific Program th , 2009 18:00 -20:00 SALONE DELLACULTURA -EXHIBITION AREA 17:00 -18:00 AUDITORIUM DELLESCIENZE Wednesday July 15 Scientific Program Stefano Piccolo,Padua, Italy TGFb signalingindevelopment andcancer Keynote lectureI Welcome Reception Chair: Francesco Argenton, Padua, Italy th , 2009 19

July 15th Scientific Program Thursday July 16th, 2009 AUDITORIUM DELLE SCIENZE

09:00 - 10:30 Session I Early Development, gastrulation, and segmentation Session Chairs: Jacek Topczewski, Chicago, USA Masazumi Tada, London, UK

1 A VEGETAL NETWORK OF TRANSCRIPTION FACTORS REGULATES DORSAL-VENTRAL PATTERNING AND GERM LAYER th SPECIFICATION IN ZEBRAFISH B. Feldman, Bethesda, USA July 16 July 2 GLOBAL CYTOSKELETAL DYNAMICS DURING TISSUE MOVEMENT IN THE ZEBRAFISH EMBRYO M. Köppen, Oeiras, Portugal

3 FILOPODIA CHARACTERIZATION IN THE ZEBRAFISH EMBRYOS L. Caneparo, Pasadena, USA

4 NOVEL FACTORS REGULATING EARLY DORSOVENTRAL PATTERNING IN ZEBRAFISH EMBRYOS Y-Y. Chen, Tübingen, Germany

5 TIME-RESOLVED ANALYSIS OF Pou5f1/Oct4 DOWNSTREAM GENE REGULATORY NETWORKS IN EARLY ZEBRAFISH EMBRYO D. Onichtchouk, Freiburg, Germany

6 AN ASYMMETRIC HER GENE REGULATORY NETWORK IN THE SEGMENTATION CLOCK C. Schroeter, Dresden, Germany

SALONE DELLA CULTURA - EXHIBITION AREA

10:30 - 11:00 Coffee break

20 12:30 -14:00 SALONE DELLACULTURA -EXHIBITION AREA 12 11 10 9 8 7 11:00 -12:30 AUDITORIUM DELLESCIENZE Thursday July 16 Scientific Program D. Gunther, Utah,USA S. Piloto,Irvine,USA MIGRATING NEURALCRESTCELLS INTRACELLULAR TRAFFICKING TO LOCALIZE N-CADHERIN IN ZEBRAFISH OVO GENESREGULATE RAB-MEDIATED M. Nowak, Dresden,Germany REGULATED BYRECEPTOR UBIQUITINATION INTERPRETATION OFFGF8MORPHOGENGRADIENTIS CALCIUM SIGNALING INZEBRAFISHLEFT/RIGHTPATTERNING K. Pooja, Singapore IN squintmRNA Srbf1, WHICH BINDS TO THE DORSALLOCALIZATION ELEMENT IDENTIFICATION OFZEBRAFISHsquintRNA BINDINGFACTOR 1, T.U. Banish,Münster, Germany BINDING PROTEIN DEADEND CONTROL OFCELLSHAPE AND CELLMIGRATION BY THE RNA F. Bontems,Génève, Switzerland BUCKY BALL ORGANIZESGERMPLASM ASSEMBLY Session Chairs: Signaling Session II Lunch th , 2009 Dirk Meyer, Innsbruck, Austria Roland Dosch, Génève, Switzerland 21

July 16th Scientific Program Thursday July 16th, 2009 AUDITORIUM DELLE SCIENZE

14:30 - 16:00 Session III Gene regulation Session Chairs: Gilbert Weidinger, Dresden, Germany Andrew Oates, Dresden, Germany

13 THE MOLECULAR MECHANISM UNDERLYING LIGHT- REGULATED PERIOD2 EXPRESSION th G. Vatine, Tel Aviv, Israel

14 CONDITIONAL GENE INACTIVATION REVEALS A CRUCIAL ROLE July 16 July FOR zgc:112094 IN ZEBRAFISH DEVELOPMENT B. Zhang, Beijing, P. R. CHINA

15 TRANSCRIPTIONAL CONTROL OF BONE FORMATION THROUGH THE REGULATORY GENE RUNX2 S. Fisher, Philadelphia, USA

16 A PHASE-ORDERED MICROARRAY SCREEN FOR CYCLIC GENES IN ZEBRAFISH REVEALS HER GENES AS THE CONSERVED CORE OF THE SOMITOGENESIS CLOCK D. Roellig, Dresden, Germany

17 A ROLE FOR ASPP2A IN ENDODERM DEVELOPMENT IN ZEBRAFISH R. Hoffmans, Boston, USA

18 CRUMBS COMPLEX COORDINATELY REGULATES NEUROGENESIS AND NEUROEPITHELIAL POLARITY THROUGH CANONICAL AND NON-CANONICAL NOTCH PATHWAY S. Ohata, Kobe, Japan

SALONE DELLA CULTURA - EXHIBITION AREA

16:00 - 16:30 Coffee break

22 24 23 22 21 20 19 16:30 -18:00 AUDITORIUM DELLESCIENZE Thursday July 16 Scientific Program ZEBRAFISH ANTERIOR CHAMBERDEVELOPMENT AND MAINTENANCE B.F. Eames,Eugene,USA C.Q. Diep,Boston,USA ZEBRAFISH KIDNEYREGENERATION RENAL STEMCELLSGIVERISE TO NEWNEPHRONSDURING M. Takamiya, Karlsruhe, Germany A. Faro, Utrecht, The Netherlands CELL POPULATION INZEBRAFISH FATE WITHIN BIPOTENTIAL HEPATOPANCREATIC PROGENITOR CDX GENESPROMOTE HEPATIC FATE AND REPRESSPANCREATIC J.K. Heath,Parkville, Australia DEVELOPMENT CELLULAR PATHWAYS DISRUPTZEBRAFISHINTESTINAL MUTATIONS INCOMPONENTSOFDIVERSEESSENTIAL DURING CHONDROCYTEDIFFERENTIATION POSITIVE AND NEGATIVE FEEDBACK FROMPROTEOGLYCANS F. Stellabotte,Los Angeles, USA JAGGED-NOTCH SIGNALING PATTERNS THE FACIAL SKELETON Session Chairs: Organogenesis Session IV th , 2009 Tom Schilling, Irvine,USA Elke Ober, London,UK 23

July 16th July 17th 24 10:30 -11:00 SALONE DELLACULTURA -EXHIBITION AREA 30 29 28 27 26 25 09:00 -10:30 AUDITORIUM DELLESCIENZE K.H. Brown, Boston,USA REARRANGEMENTS AND MARKERCHROMOSOMES FOR THE IDENTIFICATION OFINTER-CHROMOSOMAL CHROMOSOME PAINTING OFZEBRAFISHCHROMOSOMES J.K. Chen,Stanford,USA MORPHOLINO TECHNOLOGIES TRANSCRIPTIONAL TARGET DISCOVERY THROUGH CAGED R. Köster, Neuherberg,Germany EXPRESSION MAPPINGINZEBRAFISH OPTIMIZED GAL4GENETICSFORPERMANENTGENE D. Balciunas,Philadelphia,USA TRANSPOSON-BASED INSERTIONAL MUTAGENESIS PLATFORM J.M. Schaaf, Leiden, The Netherlands MOLECULE MICROSCOPY THE ZEBRAFISH AS A MODELSYSTEMFORIN VIVO SINGLE- S. Hans,Dresden,Germany DsRed2-loxP-EGFP) CONDITIONAL RED-TO-GREEN REPORTER Tg (hsp70:loxP- IMPROVED GENETICLINEAGE TRACING USING THE Session Chairs: Emerging technologies Session V Coffee break Antonio Giraldez, NewHaven, USA Reinhard Köster, Neuherberg,Germany Friday July 17 Scientific Program th , 2009 12:30 -14:00 SALONE DELLACULTURA -EXHIBITION AREA 36 bis 36 35 34 33 32 31 11:00 -12:30 AUDITORIUM DELLESCIENZE Friday July 17 Scientific Program DATA CAN BEINTEGRATED INTO THE ELECTRONICDATA STREAM AMPLIFY THE IMPACT OF YOUR RESEARCH:ENSURE THAT YOUR M. Clark,Hinxton,UK S. Sivasubbu, Delhi,India S.M. Burgess,Bethesda,USA M. Haendel,Eugene,USA H. Spaink,Leiden, The Netherlands RESPONSES INZEBRAFISH AND CARP EMBRYOS HIGH THROUGHPUT SCREENINGOFINNATE IMMUNE AND A NEWULTRA-DENSE GENETICMAP GENETIC DIVERSITYBETWEEN TÜBINGEN AND AB STRAINS, ZEBRAFISH FOR VARIANT DISCOVERY WHOLE-GENOME SEQUENCINGOF A WILDTYPE STRAINOF HAIR CELLREGENERATION IN THE ADULT ZEBRAFISH IDENTIFYING THE GENENETWORK INVOLVED ININNEREAR J. Gehrig,Birmingham,UK ENHANCER INTERACTIONS INZEBRAFISHEMBRYOS AUTOMATED HIGH THROUGHPUT MAPPINGOFPROMOTER- F.C. Wardle, Cambridge,UK T-DOMAIN TRANSCRIPTION FACTORS A MESODERMALGENEREGULATORY NETWORK DIRECTEDBY Session Chairs: Genomic workshop Session VI Lunch th , 2009 Shawn M.Burgess,Bethesda,USA Ferenc Mueller, Birmingham,UK 25

July 17th July 17th 40 39 38 37 16:00 -17:30 AUDITORIUM DELLESCIENZE 14:30 -16:00 42 41 26 AMBULACRO DELLAPITTURA AND AMBULACRO DELLALETTERATURA POSTER AREA L.Z. Zon,Boston,USA METHYLTRANSFERASE AS A NEWONCOGENE INMELANOMA IDENTIFICATION OF THE SETDB1HISTONE N. Meeker, SaltLakeCity, USA RESOURCES FORGENEDISCOVERY NEW ZEBRAFISHMODELSOF T CELLCANCER: IMPORTANT J. Neumann,Dallas,USA ZEBRAFISH A MUTATION IN ALK6B CAUSES GERMCELL TUMORS IN H. Lee,Seoul,Korea REGULATION OFMITOSIS AND CHROMOSOMEINSTABILITY ZEBRAFISH AS AN EFFICIENTMODELSYSTEM TO STUDY THE Session Chairs: Cancer C.P. Bagowski, Leiden, The Netherlands PANCREATIC CANCER 10A EXPRESSION AND BLOCKMETASTATIC BEHAVIOUR OF RETINOIC ACID RECEPTOR (RAR) ANTAGONISTS INHIBITMIR- M. Schartl, Wuerzburg, Germany FORMATION IN A TRANSGENIC MEDAKA MELANOMAMODEL GENETIC BACKGROUND DEPENDENTPIGMENTCELL TUMOR Session VII Poster PresentationODDNUMBERS Nikolaus Trede, SaltLakeCity, USA James Amatruda, Dallas,USA Friday July 17 Scientific Program th , 2009 18:00 -19:30 AUDITORIUM DELLESCIENZE 17:30 -18:00 SALONE DELLACULTURA -EXHIBITION AREA Friday July 17 Scientific Program

Chair: Nadia Rosenthal,Rome,Italy Enhancing mammalianregeneration Keynote lectureII Coffee Break th , 2009 Karuna Sampath,Singapore 27

July 17th July 18th

10:30 -11:00 SALONE DELLACULTURA -EXHIBITION AREA 28 48 47 46 45 44 43 09:00 -10:30 AUDITORIUM DELLESCIENZE A NULLMUTANT TO SURVIVE BEYOND SEXUALMATURATION A MODIFIED ACETYLCHOLINE RECEPTOR DELTA-SUBUNIT ENABLES M. Mione,Milan,Italy MEDIATED PROTEOSOMAL DEGRADATION OFONCOGENICHRAS MODEL OFCOSTELLOSYNDROMEISLINKED TO UBIQUITIN- CELLULAR SENESCENCE AND DNA DAMAGE IN A ZEBRAFISH J.M. Urban,Bethesda,USA N.S. Trede, SaltLakeCity, USA DIAMOND SYNDROME-LIKEBONEMARROW FAILURE CEYLON: A ZEBRAFISHMUTANT WITH SHWACHMAN- T.J. Carney, Singapore SYNDROME AND IDENTIFIESPOTENTIAL NOVEL DISEASEGENES GENETIC ANALYSIS REVEALS A ZEBRAFISHMODELFORFRASER M.I. Ferrante, Cambridge,UK SARCOMERIC INTEGRITYINSKELETAL MUSCLE DISRUPTION OF THE TROPONIN COMPLEXLEADS TO LOSSOF S. Oehlers, Auckland, NewZealand CHEMICAL COLITISMODELSINZEBRAFISHLARVAE Session Chairs: Disease models Session VIII Coffee break Derek Stemple,Cambridge,UK Ela Knapik,Nashville,USA Saturday July 18 Scientific Program th , 2009 12:30 -14:00 SALONE DELLACULTURA -EXHIBITION AREA 54 53 52 51 50 49 11:00 -12:30 AUDITORIUM DELLESCIENZE Saturday July 18 Scientific Program ROLE OFNOTCH SIGNALING INEARLY ZEBRAFISHCARDIOGENESIS Session Chairs: M.M. Santoro, Turin, Italy ZEBRAFISH DEVELOPMENT CHARACTERIZATION OF VASCULAR MURALCELLSDURING B.M. Hogan,Utrecht, The Netherlands ZEBRAFISH AND HUMANS SECRETED PROTEIN ESSENTIALFORLYMPHANGIOGENESIS IN THE ZEBRAFISHFULL-OF-FLUIDMUTANT IDENTIFIES A A.J. Giraldez, NewHaven, USA SIGNALING MiR-1 REGULATES ANGIOGENESIS BYMODULATING VEGF A.F. Siekmann,Muenster, Germany PATTERNING OF THE FIRSTEMBRYONIC ARTERY CHEMOKINE SIGNALING CONTRIBUTES TO REGIONAL I.M. Berger, Heidelberg,Germany FORMATION INZEBRAFISH BUNGEE - A NOVEL REGULATOR OFCARDIAC VALVE K.A. Nembhard,Durham,USA Cardiovascular system Lunch Session IX th , 2009 Stefan Schulte-Merker, Utrecht, The Netherlands Massimo M.Santoro, Turin, Italy 29

July 18th July 18th 57 16:00 -17:30 AUDITORIUM DELLESCIENZE 14:30 -16:00 56 55 30 60 59 58 AMBULACRO DELLAPITTURA AND AMBULACRO DELLALETTERATURA POSTER AREA M.V. Flores, Auckland, NewZealand T.C. Lund,Minneapolis,USA V. Wittamer, SanDiego,USA ENDOTHELIAL CELLSINZEBRAFISHEMBRYOS STEM CELLSEMERGEDIRECTLY FROMHAEMOGENIC LIVE IMAGING REVEALS THAT DEFINITIVEHAEMATOPOIETIC Session Chairs: Hematopoiesis andimmunesystem OF BONEMARROW TRANSPLANT FOLLOWING ADOPTIVE CELL TRANSFER IN A ZEBRAFISH MODEL STEM CELL(HSC)NICHE AND PLAY A ROLEINHSCHOMING SDF-1 EXPRESSINGCELLSESTABLISH THE HEMATOPOIETIC HEMOGENIC ENDOTHELIAL CELLSIN THE ZEBRAFISHEMBRYO BOTH PRIMITIVE AND DEFINITIVEHEMATOPOIESIS ARISE FROM DENDRITIC CELLS PRESENTING CELLS:BLYMPHOCYTES, MACROPHAGES AND A ZEBRAFISH TRANSGENIC MODEL TO STUDY ANTIGEN- B. Bajoghli, Freiburg, Germany CHEMOKINES AND CHEMOKINERECEPTORS THYMUS COLONIZATION ISUNDER THE CONTROLOF J.L.O. deJong, Boston,USA ZEBRAFISH MODEL TO DETECTHEMATOPOIETIC STEMCELL ACTIVITY IN DEVELOPMENT OF AN IMMUNE-MATCHED TRANSPLANTATION Session X Poster PresentationEVENNUMBERS J.Y. Bertrand, LaJolla, USA Maria Vega Flores, Auckland, NewZealand Graham Lieschke, Parkville, Australia Saturday July 18 Scientific Program th , 2009 17:30 -18:00 AUDITORIUM DELLESCIENZE Saturday July 18 Scientific Program 20:00 -02:00 CENTRALE RISTOTHEATRE Chair: Community Meeting Social Dinner Marina Mione,Milan,Italy th , 2009 31

July 18th July 19th 32 11:00 -11:30 SALONE DELLACULTURA -EXHIBITION AREA 66 65 64 63 62 61 09:30 -11:00 AUDITORIUM DELLESCIENZE FUNCTIONAL ANALYSIS OF THE HABENULA IN CONTROL OF FEAR J. Hazan,Paris, France ARCHITECTURE VIA INHIBITIONOF THE BMPPATHWAY ATLASTIN CONTROLSZEBRAFISHSPINAL MOTOR AXON M.E. Halpern,Baltimore,USA ASYMMETRY BEHAVIOURAL CONSEQUENCESOFZEBRAFISHEPITHALAMIC H. Okamoto,Saitama,Japan C. Campos,Génevè,Switzerland PROLIFERATION OF THE NEURALPRECURSORS SARA1, a TGFbeta SIGNALING ADAPTOR, REGULATES F. DelBene,SanFrancisco, USA IMAGING TECTUM BYINTERLAMINAR INHIBITIONREVEALEDBYCALCIUM SEGREGATED PROCESSINGOF VISUAL INPUTININ THE OPTIC R. Gonzalez-Quevedo, London,UK NEURONS NEGATIVE REGULATION OFNEUROGENESISBY AN FGFFROM Session Chairs: Neurobiology I:patterningandbehaviour Session XI Coffee break Marnie Halpern,Baltimore,USA Corinne Houart,London,UK Sunday July 19 Scientific Program th , 2009

11:30 -13:00 AUDITORIUM DELLESCIENZE Sunday July 19 Scientific Program 13:00 AUDITORIUM DELLESCIENZE 72 71 70 69 68 67 Session Chairs:

Neurobiology II:Sensoryorgansandregeneration Concluding remarksandend oftheMeeting E.E. LeClair, Chicago,USA VERTEBRATE TISSUE GROWTH AND REPAIR MAXILLARY BARBEL (ZMB): A NOVEL STUDY SYSTEM FOR ADULT DEVELOPMENT AND REGENERATION OF THE ZEBRAFISH M. Behra, Bethesda,USA ZEBRAFISH LATERAL LINE AFTER COPPER TREATMENT MOLECULAR EVENTSDURINGREGENERATION IN THE H. Lopez-Schier, Barcelona, Spain THE ZEBRAFISH AFFERENT INNERVATION DURING HAIR-CELL REGENERATION IN D.R. Hyde,NotreDame,USA RETINA DURING REGENERATION OF THE LIGHT-DAMAGED ZEBRAFISH REGULATING ADULT NEURONAL STEMCELLPROLIFERATION K.L. Hammond,Sheffield,UK ZEBRAFISH OTIC VESICLE THE ACQUISITION OFDORSOLATERAL CELLFATES IN THE REPRESSION OFHEDGEHOGSIGNALLING ISREQUIREDFOR B.B. Riley, CollegeStation, USA SENSORY ORGANDEVELOPMENT EARLY REQUIREMENTSFORPREPLACODAL ECTODERM AND Session XII th , 2009 Tanya Whitfield, Sheffield,UK Catherina Becker, Edinburgh,UK 33

July 19th

Abstracts 35

Talks Talks 36 TALKS SESSION I 1 Early development, gastrulation and segmentation

A vegetal network of transcription factors regulates dorsal-ventral patterning and germ layer specification in zebrafish B. Feldman, S-K.Hong, C.Levin, and J.Brown Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, USA

Two central features of early embryogenesis are the differentiation of germ layers and formation of the dorsal axis. Studies in Xenopus laevis indicate that maternal determinants in the yolky vegetal cells have a critical role for inducing mesoderm as well as the dorsal organizer. In zebrafish, factors in the yolk and yolk syncytial layer (YSL) are thought to play roles analogous to maternal determinants in Xenopus. We are exploring the functions and interactions of four transcription factors expressed in the YSL or vegetal-most blastomeres prior to germ layer differentiation or axis formation: Hnf4a, Mxtx2, Boz and Irx3a. We find that hnf4a mRNA overexpression reduces dorsal markers and recapitulates the boz loss-of-function phenotype. Consistent with this, Hnf4a depletion with morpholinos increases expression of boz and other dorsal markers. Mxtx2 overexpression similarly causes dorsal marker reduction, but also strongly induces endoderm. Intriguingly, we also observe that irx3a is expressed maternally and that its overexpression quenches the expression of mxtx2 and hnf4a alike. Thus, we are uncovering a vegetal transcriptional regulatory network involving irx3a, hnf4a, mxtx2 and boz that modulates dorsal-ventral patterning and germ-layer induction. Talks

37 Talks 38 morphogenetic during rearrangements cytoskeletal movements oftheembryo. global of importance the highlights work Together,analysis. our functional and tools imaging using hypothesis this testing currently are towards actin of transport the YSL? Athe good candidate is the microtubule network mediates spanning the vegetal half of the yolk. What We pole. vegetal the at embryo the which seals margin,ultimately EVL the shortens progressively then mesh this of contraction Local condenses. towards markedly mesh actin epiboly,the subsequent while pole vegetal the towards progress EVLcells streams pole During EVL. vegetalthe of margin the the to ordered adjacent mesh actin-rich from Together, an an YSL. generates the this actin through Simultaneously, expands divisions. then nuclear syncytium of actin-rich sequence formed, newly The boundaries. cell partial their of cell disassembly through yolk early the with during fuse blastomeres marginal initiates as stages, formation division YSL that the found of we pole imaging, vegetal confocal the live from Using originating embryo. actin of pool a and cortex structure YSL the actin from this both that suggests derives work current Our (YSL). layer syncytial yolk adramatic the within band embryo undergoes during epiboly.cell thezebrafish yolk Our spherical previous the findingsaround indicatelayer,EVL) that(enveloping EVLepithelium movementouter isthe drivenof byspreading ofdevelopment, athe contractileis actin movement such One movements. tissue hours morphogenetic of sequence a few through reorganization first the During Tissue Morphogenesis, InstitutoGulbenkiandeCiýncia, Oeiras, Portugal M. Köppen,I.Cristo,P. Almada Global cytoskeletal dynamics duringtissuemovement inthezebrafish embryo 2 Early development, gastrulationandsegmentation SESSION I During vertebrate development only short filopodia have been previously reported inthe reported previously described inzebrafishortowhatithasbeenreportedinvertebrates. been have filopodia short mouse blastocystes, with range and extension only up to 20µm and not comparable to what only development vertebrate During and cytonemes in filopodia, as gastrulation, such during urchin, sea in 50µm respectively described previously than been have nanotunneling more membrane distance cell over extending the protrusion analyzed Cell we and extensions these dynamic alongtheprojectionduringgastrulationinvivo . of the embryos persistence in and length distribution measured filopodia the andwe growth mapped their topologically Wecharacterized upto250 gastrulation. during embryo reaching zebrafish offilopodia developing the presence the time first the for describe We examples, ofshortrangemembranecellprotrusionsaremicrovilli,lamellipodiaandruffles. Wellanimals. studied among share allowed and ubiquitous are mechanisms these communication, cell cell- for approach used and behavior exploratory of This characteristic are example, for resolution.protrusions, function. Cellular cells the in cell changes of indicators often, morphology, and single shape cell of visualization with cytoarchitecture, gastrula the of availability the and movement gastrula the the allows embryo, the of clarity optical diversity. the cells zebrafish, In movements are required, to set morphogenetic up the and correct events formation of signaling the conserved body well plan and of to series guarantee future a gastrulation, vertebrate During Biology ImagingCenter, CaliforniaInstituteof Technology, Pasadena, USA L. Caneparo,P. Pantazis,S.E.Fraser Filopodia characterization inthezebrafish embryos Early development, gastrulationandsegmentation SESSION I , in the imaginal disc Drosophila and in different cells types in the vertebrate immune system. multiplex fluorescent tags consent to highlight to consent tags fluorescent multiplex n vivo; in 3 visualization of visualization vivo in e quantitatively we i lnt in length in µm n vivo in we 39

Talks Talks 40 axis oftheEVL isspecifiedindependentlyofknowndorsoventralsignalsinearlydevelopment. and Max in early embryonic patterning and suggest a novel mechanism whereby the patterning of the EVL. Our analyses have revealed the roles of maternally expressed Sox19b the dorsoventral dorsalise the embryos indicating that they have of a over-expressionfunction in embryonic Furthermore, patterning in addition EVL. to dorso- the the of alter polarity knockdown, ventral morpholino or over-expression by either levels, expression max in perturbations that show We proteins. interacting promoter minCrestin as (Max) a yeast-one-hybrid screen and identified members of the Sox familythat and Myc-associated factor-Xpathway early an is regulates the dorso-ventral patterning of EVL. Tothere isolate these novel components, we performed that indicates This morpholinos. using signaling Chordin-BMP early the at cells EVL sphere stage. dorsal Intriguingly, labels the domain fortuitously of which GFP expressing line cells did transgenic not change minCrestin::GFP when we a perturbed used we issue Tothe patterning. address its in involved are cues patterning dorsoventral known the Spemann’s of organizer. equivalent zebrafish It much is the not stage, is clear whether which this 256-cell shield, cell embryonic layer the exhibits the dorsoventral of atpatterning at formation all differentiates the and before whether EVL The layer. cell protective outermost the (EVL), layer enveloping the from derived are cells peridermal the ectoderm, embryonic from develops bi- is epidermis larval layered and consisting of embryonic outer the peridermal zebrafish, ectoderm and inner In embryonic basal signaling. epidermal BMP from cells. of While formed the influence basal the is epidermis under epidermis The antagonizing signaling. the BMP by and development Chordin of embryonic activities vertebrate during early very established is axis Virtually all animals have left-right, anterior-posterior, and dorsoventral polarity. The dorsoventral Max-Planck InstituteforDevelopmental Biology, ofGenetics, Department Tübingen, Germany Y-Y. Chen,M.Sonawane,Harris,andC.Nüsslein-Volhard Novel factorsregulatingearlydorsoventral patterninginzebrafish embryos 4 Early development, gastrulationandsegmentation and knockdown of knockdown and 19b sox SESSION I and sox19b Max development contributes to the effort of producing defined cell fates from pluripotent stem pluripotent from fates cell defined producing cells. of effort the to contributes development zebrafish from gained knowledge the that envision we vertebrates, across conserved are model Oct4/Pou5f1 of modules totipotency as control in ES cells in mammals. Since the Pou5f1 downstream reimplicated genes involved in our were subnetworks control pluripotency embryonic Oct4/Pou5f1 ancient evolutionary the of of elements model properties the that dynamic Wesuggest, motifs. The the network certain other.to tissue the neural the on in differentiation cells of timing differentiating the link and to us side, allows one the series, on cells time progenitor neural to combined We targets. mathematical blastomers predictive totipotent of segregation the during operates which model, network gene-regulatory a transcriptional build to data Sox2 hybridisation in-situ and direct experiments overexpression distinguish to experiments Pou5f1 – Sox2 (SoxB1) dependent transcriptional network and performed additional microarray neuroectoderm, (ectoderm, stage midgastrula mesendoderm, respectively). Weat foccused patterns our efforts in spatial characterising the neuroectodermal unique part in expressed zebrafish them of in mass, each , FOXD3 cell and Sox2 Klf4, include vertebrates inner across conserved Pou5f1 Transcriptionalzebrafish of targetssubnetworks. the separated spatially of several of population upstream operate cell to seems Pou5f1 stem homogenous the in pluripotency RT-PCR.time real or We show, controls Pou5f1 where embryos, mammalian to contrast in that suppression of translation the of by primary experiments target additional gene in mRNAs, distinguished results validates were targetsby in-situ Oct4/Pou5f1 hybridisation Direct development. organism multicellular in involved genes other and factors transcription for enrichment strong revealed expression developmental and function Ontology Gene of Analysis mutants. Pou5f1 that targetswere not expressed Oct4/Pou5f1 or showed either advanced, of delayed or deregulated groups expression in Oct4/ identified data series time of analysis combined A microarrays. epiboly) in zebrafish wild type and Oct4/Pou5f1 deficient embryos in 1 hr intervals using Agilent 75% to egg unfertilised (from time profiles expression gene zygotic early and maternal generated of data series have we Pou5f1/Oct4 of downstream networks transcriptional the of function endoderm, epiboly defect and BMP pathway deficiency. To further understand the evolution and The MZspg mutant phenotype includes several independent defects described so far: absence of been has established: function MZspg Pou5f1 mutants, for model which genetic are a devoid zebrafish, of In both controversial. maternal remains vertebrates and zygotic expression of Pou5f1. lower of development the in role Pou5f1/Oct4 role this of conservation however,evolutionary The Pou5f1/Oct4 transcription factor controls pluripotency in mammalian embryonic stem cells, Freiburg, Germany 1 W. Driever D.Onichtchouk embryo Time-resolved analysis of Pou5f1/Oct4 downstream gene regulatory networks in early zebrafish Early development, gastrulationandsegmentation SESSION I nvriy f riug Febr, Germany; Freiburg, Freiburg, of University eeomna booy Uiest o Febr, riug Germany; Freiburg, Freiburg, of University biology, Developmental 1 1 , F., Geier 2 , B. Polok B. , 1 , R. Moessner R. , 3 oeua Ebylg, P fr Immunobiology, for MPI Embryology, Molecular 1 , D. Messerschmidt D. , 3 5 , V., Taylor 2 nttt o Physics, of Institute 3 , J. TimmerJ. , 41 2 ,

Talks Talks for the for We use multiple time-lapse microscopy to measure somitogenesis period in three novel different mutants of these of oscillator components. map studies interaction the reinvestigate to us morpholino-knockdown prompting (2), with inconsistent are previous genes her However, in gene phenotypes. any similar qualitatively of segmentation to lead should mutation genes different upon of mutation changes (2) that imply period and network somitogenesis the (1) that predict models These loop. feedback autoregulatory negative delayed a through oscillations generate that genes her interacting multiple of network symmetric a propose mechanism oscillator core the of models display transcriptional oscillations in the PSM and somites. are the into important components mesoderm of of paraxial Members the the clock. of Current subdivision sequential the presomitic controls unsegmented (PSM), the mesoderm in operating oscillator transcriptional a clock, segmentation The Genetik, Universitaet Koeln, Germany 1 42 C. Schroter An Asymmetric herGeneRegulatoryNetwork intheSegmentationClock embryos mutant for the in decay they whereas stages segmentation throughout persist oscillations mutants, single hes6 the In pace. type wild at segment mutants two other the while period, somitogenesis rescues the decay of the oscillations seen upon mutation of the the Strikingly,crossing oscillations. abrogates neatos n h cokok n sol bodn u udrtnig f h gnrto of generation the of understanding our broaden should and developmental rhythms. clockwork the in interactions important have heterodimers her7/hes6 and biochemical of predictions testable makes model network genetic This functions. opposing and her1/hes6 that proposes but heterodimers, her7 defines a novel segmentation phenotype. Our model does not rely on her7 homodimers segmentation or her1/ mutant different regulatory the explain phenotypes. genetic Furthermore we to can clock’sexplain the oscillator interactions period change in segmentation the asymmetric on the builds of that modelnetwork mathematical minimal a develop We towards asymmetricinteractionsbetweenthem. the of functions distinct indicate findings these Max Planck Institute of Molecular Cell Biology and Genetics,Germany;Dresden,and Biology Cell Molecular of Institute Planck Max 6 her1, 1 , M.Gajewski and her7 aiy f rncitoa repressors transcriptional of family (her) related split of enhancer and hairy genes. Unexpectedly,genes. hes6 of mutation only her7 gene. Combined loss of 2 , A. C.Oates 1 Early development, gastrulationandsegmentation mutation into the into mutation hes6 genes in the segmentation clock and point and clock segmentation the in genes her her1 and her7 her7 or her1 leads to an increase in increase an to leads hes6 gene alone. Taken together, mutant background mutant her7 and hes6 completely hes6 mutant, which SESSION I 2 Institut fuer Institut and her1 1 F. Bontems Bucky ballorganizesgermplasmassembly Signaling SESSION II gene, we positionally cloned the cloned positionally we gene, the identify Tomolecularly assembly. plasm germ for essential is Buc that demonstrating We show that in unknown. the fertility of the future embryo, the key regulators of Balbiani body assembly in vertebrates are for critical is plasm germ Although humans. including throughthe animals many of oocytes in has found been which body, Balbiani the structure, development cytoplasmic distinct a forming the duringembryonic oogenesis during of assembles stage first plasm Germ plasm. ofgametes germ termed determinants maternal of localization the formation specifies Zebrafish of Cell andDevelopmental Biology, University ofPennsylvania, Philadelphia, USA biochemical functionandcellbiologicalroleofBucduring Balbiani bodyformation. the study to tool good a be will also but vivo, in localization plasm germ reports only not line This stage. oblong until cells plasm germ primordial germ four the in the later and to furrows cleavage localized first the is at Buc-eGFP stages embryonic early the At oocyte. the of cortex protein Buc reports eGFP-fluorescence the localization addition In activity. reflects Buc transgene endogenous the the that of indicating most phenotype mutant the rescues line This line. Buc-eGFP transgenic a established we localization, body Balbiani in Buc of role Tothe understand better represents thefirstregulatorofgermplasmformationinvertebrates. discovered that evidence providing embryo fertilization. Interestingly, overexpression of Buc-eGFP fusion localizes to the Balbiani body and in the germ plasm of the early embryo after to aggregate germ plasm in the oocyte . Consistent with this result, we discovered that assay in an zebrafish stageinjected I oocytes. Using this assay we show that Bucculture proteinand activityinjection isan necessary developed we Buc, of body.function ToBalbiani the the characterize in initiating pattern localization dynamic novel a displaying ovary the in expressed exclusively Departement Departement de Zoologie et Biologie Animale, Université de Génève, Switzerland; 1 , A. Stein homologs in many genomes including mammals our results suggest that Buc that suggest results our mammals including genomes many in homologs buc , which we discover in the Balbiani body and later during oogenesis to the to oogenesis during later and body Balbiani the in discover we which vivo , in bucky ball ( 1 , F. Marlow buc) mutant females the formation of the Balbiani body is disrupted 2 is sufficient to recruit germ plasm components. Since we Since components. plasm germ recruit to sufficient is buc , J.Lyautey mutation. buc buc generates additional germ cells in the zebrafish 1 , T. Gupta encodes a novel protein and its mRNA is mRNA its and protein novel a encodes Buc 2 , M.C.Mullins 2 7 ,R.Dosch 2 Departement Departement 1 buc 43

AbstractTalks AbstractTalks 44 (e.g. RNAs specific protecting by functions Dnd that found We and survival. maintenance fate cell motility), of acquisition the (e.g. behavior cell in defects severe in results of Knockdown granules. perinuclear the to localized is it where PGCs, in expressed end ( dead provided the by encoded is zebrafish onmaternally in component germ-plasm maternally-provided such important An depends (PGCs) cells line andacquirepropercellbehavior. germ primordial determinants. These determinants, collectively termed germ-plasm direct cells to enter the germ zebrafish of Specification Institute ofCell Biology, ZMBE, Münster, Germany T. U.Banisch, M.Goudarzi, E.Raz Control ofcellshapeandmigration by theRNA bindingproteinDeadend The identification and functional analysis of such RNAs that are involved in controlling cell incontrolling areinvolved that RNAs such shape andmotilitywillbedescribed. of analysis functional and identification The Dnd aswellRNAsthatarepotentiallyregulatedbythecomplex. To determine the molecular basis for Dnd function we have identified proteinsby that interact with process a cells, germ which thesameRNAsareinhibitedinsomaticcells. the in degradation RNA and inhibition translational RNA–mediated 8 ) gene. The Dnd protein contains an RNA binding domain and is specifically is and domain binding RNA an contains protein Dnd The gene. dnd) and tdrd7 ) from micro from nanos1) SESSION II Signaling dead end dead 1 Singapore; and localizessqtRNA todorsal. the other factors that bind to sqt RNA, and understanding the mechanisms by which Srbf1 binds purifying at aimed are efforts Current defects. embryonic to leads embryos in Srbf1, type wild by TILLING, and characterization is in progress. Interestingly, over-expression of mutant but not and of necessary for is binding to Analysis sqt RNA. An Srbf1 ENU-induced mutation in the protein. of N-terminus acid-binding the that nucleic sufficient shows for binding to sqt RNA. a We have also Srbf1 identified key residues in Srbf1 that are essential in is mutations Srbf1 point and sequence. deletions peptide its determined and chromatography, by extracts oocyte from Srbf1 purified sqt have Wethe element. to localization binds dorsal (Srbf1), 1 factor binding sqt-RNA these, of One 3’UTR. sqt the within regions distinct to specifically bind which factors RNA-binding and multiple show gel-shift assays RNAcross-linking UV structure. secondary conserved potential a and blocks, conserved multiple revealed species cyprinid dorsal ~30 from 3’UTR sqt in the of footprinting Phylogenetic 3’UTR. sqt step the early an is of nt 50 first the within lies element localization dorsal the and factors, localization dorsal for sufficient other possibly and necessary is (UTR) 3’region sqt untranslated and The 2005). , Nature al., et (Gore RNA, specification sqt that suggests of This dorsal. localization embryonic the asymmetric predicts and stage, 4-cell the by cells 2 to localized asymmetrically is (Sqt) Squint factor Nodal-related the encoding mRNA maternal that showed previously We embryos. zebrafish in axis embryonic the of specification investigating are We Identification of Zebrafish squint RNA Binding Factor 1, Srbf1, which binds tothedorsal binds K. Pooja which Srbf1, 1, Factor localization elementinsquintmRNA RNABinding squint Zebrafish of Identification Signaling SESSION II eae Lf Sine Lbrtr, Singapore; Laboratory, Sciences Life Temasek 1 , P. C.Gilligan 3 Max PlanckInstituteofCell BiologyandGenetics, Dresden, Germany 1 , R.Philp 2 , S.Winkler 3 , K.Sampath 2 ipoesn Tcnlg Isiue Biopolis, Institute, Technology Bioprocessing srbf1 locus has been recently obtained 1 9 45

Talks AbstractTalks 1 establishment oftheleft/rightaxis. downstream of fluid flow in KV and upstream of the asymmetric gene expression cascade in the mobilization calcium for requirement the place experiments These axis. the of side right the on free calcium levels on the right side of KV is sufficient to drive left-side specific gene expression intracellular of elevation that show we experiments, additional in In step patterning. required left-right normal a is KV the of side left the on mobilization calcium intracellular that indicate experiments flow. these fluid Together or ciliogenesis structure, KV affect not does function left-side- or Second, flow. fluid of time in of flow expression fluid inhibition RyR on Third, depends the mobilization KV. calcium specific at mid-somitogenesis, during required is patterning left-right in function RyR that indicate studies inhibitor pharmacologic First, expression: gene Calcium release appears to be part of the signaling pathway between fluid flow and asymmetric along with left-side-restricted patterns is of gene expression and subsequent function organ morphogenesis. or expression RyR When blocked, KV, function: is widely, the calcium in inhibited of specifically or mobilization left-side-specific RyR on depends Vesicle. Kupffer’s calcium of of side mobilization left the The on specifically elevated are levels calcium intracellular free left-right of cascade entire of the embryos, zebrafish elevation of stage somite RyR5-8 on the dependent function. At calcium intracellular left-side-specific that requires show morphogenesis organ we and Here expression well. gene asymmetric as processes other in they involved suggesting types are cell non-muscle many in expressed widely are RyRs However skeletal are and muscle. cardiac that vertebrate in channels coupling contraction release excitation in calcium role their intracellular for known of best class one are (RyRs) Receptors Ryanodine asymmetric geneexpressionandsubsequentleft-rightpatterning. mobilization of intracellular calcium is dependent on normal fluid flow in KV and is required for left-side-specific that show we Here arises. expression gene side-specific left where mesoderm, understanding of how the information of fluid flow is transmitted from the KV to little the lateral have plate still zebrafish. We in asymmetries left-right to leads that expression gene of cascade Directional fluid flow in Kupffer’s Vesicle (KV) is required for the initiation of the left side-specific Utah, USA 46 D. Gunther Calcium SignalinginZebrafish Left/Right Patterning Departments of Human Genetics and and Genetics Human of Departments 10 1 , M.Jurynec 1 , J. Amack 2 , H.J. Yost 2 Department of Neurobiology & Anatomy, Neurobiology of Department of University 2 , D.J.Grunwald 1 SESSION II Signaling optn cmatet ad hs s n motn mas o nope opoe gradient morphogen uncouple to means important formation andinterpretation. an is thus and compartments competent endocytosis and sorting determines the residence time of receptor-ligand complexes its in signaling for sufficient is compartments endosomal clearance from the extracellular space early and thus gradient formation. In contrast, and regulation of Fgf surface cell the to Fgf8 of recruitment that Wepropose interpretation. gradient altered to due are indicating changes signaling observed gradient the that morphogen Fgf8 extracellular the on effect no has ubiquitination receptor Fgf impaired that show we a Spectroscopy Correlation Fluorescence Using embryos. zebrafish in to leads receptors Fgf of the of function ubiquitination the broadening reduced of Fgf that with target gene show expression interfering and we a delay Cbl of By Fgf8 endocytosis ligase and elusive. lysosomal ubiquitin transport remains the tissues target within interpretation their how into gained been has however,maintained, and govern formed mechanisms are cellular gradients which morphogen insight much Recently, gradients. morphogen of interpretation determined largelyby are decisions fate cell organogenesis and development embryonic During Biotechnology Center andCenter forRegenerative Therapies, CRTD; TU Dresden, Germany M. Nowak, R.S. Yu, A. MachateandM.Brand Interpretation ofFgf8morphogengradient isregulatedby receptorubiquitination Signaling SESSION II 11 47

Talks Talks 48 Ncad inmigratingNCcells. embryo and early these results the suggest that Ovo proteins regulate Rab-mediated in intracellular trafficking toTaken localize together morphants. Ovo NC cells in development rescued function in their of inhibitors specifically chemical expressed were genes Rab these of several Ovo morphants were members of the Rab family involved in act intracellular as transcriptional trafficking.repressors, and we found that Surprisingly,some of the most highly upregulated genes in with Ncad and depletion causes a decrease in Ncad at the cell membranes. Ovo genes typically of migration for required are pigment relative progenitors, in part by close regulating the secretion of its N-cadherin. Ovo interacts genetically and Ovo gene target Wnt a that show we Here but alsoplayaroleinthespecificationofNC-derivedpigmentcells. has suggested that secreted Wnts surfaces, induce these changes in adhesion in NC, their leading to migration, on cadherins of evidence Experimental cadherin-6/7. upregulating expression and E-cadherin and N- both downregulating the regulating by initiate behaviors cells migratory (NC) highly crest their neural Vertebrate adhesion. cell regulated dynamically by controlled are cells of behaviors migratory how is biology developmental and cell in issue Afundamental University ofCalifornia-Irvine, USA S. Piloto,T. Schilling in migrating neural crestcells n-cadherin localize to trafficking intracellular rab-mediated regulate genes ovo Zebrafish 12 SESSION II Signaling 1 G. Vatine The molecularmechanism underlyinglight-regulatedperiod2expression Gene regulation SESSION III Light induces the expression of expression the induces Light Light exposure is mandatory for the onset of the pineal circadian clock that drives these rhythms. earliest detected circadian rhythms in zebrafish, appearing as early as days 2-3 of development. Daily rhythms of the melatonin hormonal signal and of gene expression in the pineal gland are the Toxikologie undGenetik, Forschungszentrum Karlsruhe, Germany of how clock gene regulation has evolved in vertebrates as well as of general photic cellular photic general of as well responses. as vertebrates in evolved has regulation gene clock how of understanding general a to contributes and clock circadian the of light-entrainment underlying of regulation clock enables light which in mechanism hierarchic a suggest studies these of findings The tested. functionally and vitro the of order to understand the mechanism underlying light-induced in the pineal gland but also throughout the body and in cell lines, by an unknown mechanism. In pineal circadian clock. Interestingly, in zebrafish, light exposure induces responsible for its clock-regulated expression. Using is expression. element clock-regulated E-box its an for while responsible expression, light-induced its for critical the is within enhancer box that D revealed a analyses LRM, Functional cells. zebrafish in expression gene of regulation light direct confer to counterpart zebrafish its throughout for substitute can conserved LRM human the highly and evolution is sequence LRM the Interestingly, induction. light for sufficient the in (LRM) module light-responsive a revealed clocks. circadian light-entrainable directly contain to known are which cells reporter constructs in zebrafish embryos and by means of stable transfection of PAC-2 zebrafish Department of Neurobiology, Faculty of Life Sciences, Tel Aviv University, Israel; University,TelAvivIsrael;Sciences, Life of Neurobiology,Faculty of Department techniques, transcription factors that bind to these elements were identified, characterized identified, were elements these to bind that factors transcription techniques, promoter was performed by transient and transgenic expression of expression transgenic and transient by performed was promoter per2 1 , D.Vallone 2 , N.S.Foulkes period2 ( . This study extends the understanding of the mechanisms the of understanding the extends study This per2. 2 , Y. Gothilf ) which is, in turn, important for the onset of the of onset the for important turn, in is, which per2) 1 promoter which is both necessary and necessary both is which promoter per2 approaches complemented by complemented approaches vivo in per2 expression, functional analysis 13 per2 expression not only This analysis has analysis This promoter- per2 2 Institut für Institut 49 in

Talks Talks 1 suggest that suggest present within the lateral mesoderm, while blood cell development remained normal. These data severely affected. Angioblasts were reduced and failed to migrate to midline although they were including RNA 22hpf. after somites and heart to restricted became and detectable maternally was gene this of Expression ligase. E3 of construct in the first trap intron gene conditional of the of the insertion the gene by caused was phenotype mutant the that confirmed with insertion the reversing by one was identified as a conditional mutant with embryonic lethal phenotype only beingR-SAinsertions, the introns. intron Among in apparent 15 and exons in 7 obtained we insertions, unique Cre 94 mapping From insertion. the reversing by phenotype the rescue to splicing, used be can recombinase abnormal causing (F-SA), transcription of direction same the Similarly,in expression. is insertion gene the endogenous if the disrupt to cassette trap gene the invert to applied be can recombinase Cre (R-SA), gene the to opposite being sequence SA the of orientation the with introns its of one in interest of gene a hits insertion the When transcription. stop to signal contains unidirectional Cre/loxP a system using coupled zebrafish in with trap a gene splice for screen acceptor pilot (SA) a performed and have Wepoly-adenylation 2 may berelatedtocancerandhumanvasculardiseases. wide screen of mutations. The potential role of loxP mediated conditional mutation in zebrafish and the method could be scaled up for genome 50 For Role Crucial a B. Zhang Reveals InactivationDevelopment Gene Conditional Los Angeles, USA Genetics, College of Life Sciences, Peking University, of Ministry Education, Beijing, P. R. CHINA; Department of Molecular, Cell & Developmental Biology, University of California Los Angeles,Los California Biology,of Developmental University Molecular,& of Cell Department and Biology Developmental of CenterDifferentiation, and Proliferation Cell of KeyLaboratory 14 1 , W. Liang flk1, is required for vasculogenesis. To our knowledge, this is the first Cre/ first the is this Toknowledge, vasculogenesis. our for required is zgc:112094 and etsrp . 1 , J.Ren , showed that blood vessel formation in the mutant embryos was embryos mutant the in formation vessel blood that showed , gata1 1 , X.Ren hybridization analysis with markers for different tissues, different for markers with analysis hybridization situ in RA neto. TPR n mN rsu experiment rescue mRNA and RT-PCR injection. mRNA Cre 1 , M.Liu zgc:112094, which encodes an ASB-5 homologous protein 1 , Z.Zhu zgc:112094 in cell migration and/or proliferation 1 , andS.Lin 1, 2 n Zebrafish in zgc:112094 based construct that construct Tol2based Gene regulation SESSION III 1 S. Fisher Transcriptional controlofboneformationthroughtheregulatorygeneRunx2 Gene regulation SESSION III opportunity toexamineregulationofRunx2 indetail. species. Thus our identification of this enhancer, and the transgenic fish carrying it, afford us the across conserved be also will it upon acting factors regulatory the that anticipate we zebrafish, earliest the regulate signals stages of what bone formation extension in the embryo. by Given the conserved and function enhancer,of the the human enhancer in regulate factors what about mediating sites bone–specific regulation of binding factor transcription represent hypothesize we which sequences, short multiple zebrafish from a with associated sequences one analyzed including orthologous vertebrates, to have comparison We in growth.enhancer human later the of and sequence the embryogenesis throughout formation bone ongoing picture dynamic of and detailed a the gaining are of we enhancer, analysis the Through by regulated bone. pattern to expression specific is and expression, cartilage detectable regulate not are expressed prominently during cartilage development. However, this enhancer element does In contrast to mouse to contrast In weeks. several for bones forming newly in transiently expressed be to continues and skeleton, enhancerdrives the craniofacial the of bones promoter, forming earliest minimal the in fertilization post days aheterologous 3 before expression GFP with conjunction In zebrafish. the in element from the human transcriptional regulation of the of mechanisms specific the skeletogenesis, in importance its Despite well. as conserved be will Runx2 of role functional critical the that suggests formation bone early in orthologues zebrafish that of other known markers of osteoblast differentiation. The conserved expression pattern of the The transcription factor Runx2 is required for bone formation in mouse, and its expression Molecular MedicineGraduateProgram,precedes JohnsHopkinsUniversity, Baltimore, USA Cell and Developmental Biology, University of Pennsylvania, Philadelphia, USA;Pennsylvania,Philadelphia, of Biology,University Developmental and Cell 1 ,M.Gallagher1,C.H.Lee , which shows limited expression in cartilage, the zebrafish genes zebrafish the cartilage, in expression limited shows which Runx2, RUNX2 gene that regulates osteoblast–specific reporter gene expression Runx2. Based on this information, we are testing specific predictions Runx2 gene itself are not known. We have identified an enhancer 2 gene. We find multiple conserved multiple Wefind gene. runx2 15 2 Cellular and Cellular 51

Talks Talks O. Pourquie in zebrafish using a phase-ordered microarray screen. We identifed the known cyclic genes, cyclic known components, new the two discovered and identifed We screen. microarray phase-ordered genes a cyclic using find zebrafish to aimed in we network, genetic this of conservation the Toassess repressors. transcriptional of family HES/her the of targetgenes downstream and pathway signalling Notch Delta/ FGF the to restricted and are zebrafish Toin 2006). genes Wnt al., cyclic et date, (Dequeant Delta/Notch, signalling with associated genes cyclic thirty than more are there mouse, In mechanism ofthisoscillatingnetworkarenotwellunderstood. forming somite boundary. Despite its importance for the vertebrate body plan, the evolution and cyclic genes form a segmentation clock in the PSM that defines timingof body loops feedback Oscillating somites. into and(PSM) mesoderm presomitic unsegmented from embryonic axis positionvertebrate of eachthe newly of segmentation sequential and rhythmic the is Somitogenesis 1 52 asthe genes her D. Roellig reveals zebrafish in genes cyclicconserved coreofthesomitogenesisclock for screen microarray phase-ordered A S, iia t te te Hs sbfml gns Wees vrepeso of over-expression Whereas genes. sub-family Hes5 other the to similar PSM, and clock, segmentation zebrafish the of components core known to it compared and genes, characterized we this answer To somitogenesis. of aspects different in roles independent play or redundantly function they whether question the genes her Eleven unclear. remains somitogenesis oscillate in in zebrafish genes posterior PSM, in HES/her contrast to four of known in mouse. role This large number the raises Nevertheless, different of clockworks complex species. the of ancestral unit the minimal of the form core today the and genes were clock, HES/her segmentation genes HES only that examined, suggesting expression, vertebrates cyclic all conserved across show Thus counterpart. simpler be mammalian may its clock segmentation than zebrafish the that suggesting found, were pathways Wnt causes weak disturbances, weak causes the mouse. as such clocks slower in required not function a zebrafish, the of clock segmentation fast the to robustness conferring loop oscillatory an form members sub-family Hes5 duplicated the which for role a indicate data these conclusion, In genes. Hes5 cyclic all of knockdown by tested is hypothesis a in affected not of function of Loss MPI-CBG, Dresden, Germany; 16 . In contrast to contrast In her7. 1 in somitogenesis that is distinct from known her genes. We propose a model in model a Wepropose genes. her known from distinct is that somitogenesis in her2 , A. Kumichel 2 mutant, suggesting that other Hes5 family members compensate. This compensate. members family Hes5 other that suggesting mutant, her2 or her1 1 and her1 , A. Krol over-expression results in strong disruption of somitogenesis. of disruption strong in results over-expression her2 yield distinct somitogenesis phenotypes, but somitogenesis is somitogenesis but phenotypes, somitogenesis distinct yield her7 2 Stowers InstituteforMedicalResearch, Kansas City, USA 2 , ML. Dequeant , cyclic expression of expression cyclic her7, and her2 . Strikingly, no cyclic members of FGF or FGF of members cyclic Strikingly,no her4. 2 , O. Tassy her2, n o nn cci Hs sub-family Hes5 cyclic nine of one 2 , G. Hattem is restricted to the posterior the to restricted is her2 2 , E. Glynn Gene regulation SESSION III or her1 2 , A. Oates her1 her7 1 , or Bonandif Aspp2a canbefoundonthepromoterofcas. induce ih o ad a b isl induce itself by can and Bon with and mezzo R. Hoffmans A rolefor Aspp2a inendodermdevelopment inzebrafish Gene regulation SESSION III 1 expression of expression the bind and Eomes of the transcription factors vertebrates. in formation Nodal endoderm signaling in results determinant in major phosphorylation the and is activation pathway of TGFbeta/Nodal The Smad2, which controls the expression development. pancreas and liver in defects in results turn in which formation, endoderm affects loss aspp2a that suggests This embryos. injected control to compared embryos injected morpholino aspp2a expressing cells. Epistasis analysis showed that in reduced severely is of expression the is and can be marked by marked be can and in reduced However, and neuroectoderm (sox2) are unaffected (bmp2b) in the mesoderm/ectoderm ventral (ntl), mesoderm presumptive for Markers analyzed. in the formation of the three germ layers the expression of marker genes during gastrulation were that are not seen in control morpholino injected embryos. To characterize the function of both maternally and zygotically. five tumors. Here we used the zebrafishco- totranscriptional studya theas rolerole activator of and its binding for partner known of p53, best a is tumor suppressor p53) gene of that is Protein mutated Stimulating in (Apoptosis half ASPPof all human Dana Farber CancerInstitute, Boston, USA; family members. In this work we focused on focused we work this In members. family aspp expression. Current efforts focus on testing if Aspp2a binds directly to Gata5, Eomes Gata5, if to testing directly on binds focus Aspp2a efforts Current expression. cas xrsin wih ak eddra cls n frrne cls i severely is cells, forerunner and cells endodermal marks which expression, sox17 injected embryos. The liver and pancreas are endodermal derived organs derived endodermal are pancreas and liver The embryos. injected aspp2a . These results indicate that indicate results These cas. 1,2 and sox17 , S.Sidi and bon rmtr n induce and promoter cas 1 and A. T. Look injected embryos and there is also a slight reduction in reduction slight a also is there and embryos injected aspp2a and hhex . In foxa2. , gata5 . However the number of number the However mezzo. aspp2a morpholino injected embryos show at 1 dpf tail defects bon and . The expression of both of expression The ins. morphants Smad2 phosphorylation is unaffected as unaffected is phosphorylation Smad2 morphants aspp2a xrsin Te rncito factor transcription The expression. cas 1 mezzo. Bon and Gata5 form a complex together with aspp2a morphants compared to uninjected controls. functions together with together functions aspp2a 2 Hubrecht Institute,Hubrecht Utrecht, Netherlands aspp2a is epistatic to xrsin Mzo ucin redundantly functions Mezzo expression. cas aspp in development. In zebrafish there are , which is ubiquitously expressed ubiquitously is which aspp2a, and hhex and foxa2 gata5 and 17 was reduced in the in reduced was ins expressing cells expressing cas and gata5 euae the regulates cas eomes but not to to eomes aspp2a gata5 53

Talks Talks and morphologythroughtheregulationofNotchactivity. NICD. These results may suggest that Epb4.1l5 cooperatively regulates neuroepithelial functions mutant, and knocking down of the down knocking rescued and mutant, partially adhesion, and polarity cellular regulate and NICD of downstream In addition, over-expression of constitutive active form of R-Ras, which had been reported to act mutant, and over-expression of the Notch intra-cellular domain (NICD) rescued the the in reduced significantly Interestingly,was over-expressionCrumbs2. activity of Notch Notch activator -Secretase in fruit fly.the regulate negatively The and Epb4.1l5 by regulated negatively be to reported of been had proteins downstream acting molecules potential analyzed next we level, how Epb4.1l5 cooperatively regulate morphology.neuroepithelial Toreveal and functions and functions morphology neuroepithelial in of the control molecular cooperative the for required is vagus motor neurons were also affected in the holm mutant. These results suggest that addition, neuroepithelial functions such as neurogenesis and guidance for tangential migration of junctions, and the tight extension junctions, of apical adherens polarity,processes to apicobasal ventricular surface as were such disrupted in morphology the Neuroepithelial mosaic eyes), whichencodesaputativeadaptorprotein. gene, polarity neuroepithelial apicobasal polarity, we as analyzed the zebrafish such neuroepithelial morphology their and which neurogenesis as such by functions their mechanisms regulate coordinately cells molecular the understand To important. crucially are processes, apical a of of functional establishment nervous the tip system, For the cooperative formed. the control are of junctions neuroepithelial At tight functions, respectively.and and morphology adherens surfaces, as neural such pial junctions cellular and of ventricular guidance the to neurogenesis, extend as that such development, neural migration, of and axon aspects guidance. They various have very control unique morphology with apical cells, stem and neural polarized highly are basal which cells, neuroepithelial processes development, neural the In 1 I. Masai 54 S. Ohata Canonical andNon-canonicalNotch Pathway through Polarity Neuroepithelial and Neurogenesis Regulates Coordinately Complex Crumbs RIKEN BrainScienceInstitute, Japan; 18 2 , andH.Okamoto 1 , R. Aoki 1 , S. Kinoshita 1 (erythrocyte protein band 4.1-like 5, formerly known as known formerly 5, 4.1-like band protein (erythrocyte epb4.1l5 1 canceled the recovery of the of recovery the canceled r-ras , S. Tsuruoka-Kinoshita 2 Okinawa InstituteofScienceand Technology, Japan holm mutant phenotype was partially phenocopied by holm mutant that has a mis-sense mutation in the 1 , M. Yamaguchi mutant phenotypes by phenotypes mutant holm Cub family Crumbs epb4.1l5. 2 , H. Tanaka Gene regulation holm mutant. In SESSION III holm mutant. 1 , H. Wada epb4.1l5 holm holm 1 , regulated byJagged-Notchsignaling. arches during patterning stages. Currently, we are pharyngeal testing whether the these of candidate genes domains are discrete also within expressed are that genes candidate present Wewill precursors. skeletal of domains DV different in GFP expressing zebrafish transgenic from cells morphology, (FACS) sorting cell activated fluorescent performed we of analysis microarray and rescued. mutation in mutation as such genes, specifier activating by identity jaw upper versus lower specifies morphogen (Edn1) Endothelin1 the of gradient dorsal clear.to less ventral A are identity dorsal for arches.responsible however,genes pharyngeal well-conserved the along the skeleton facial the dorsoventral axis. Several genes of have patterning been identified that arein required for a ventral skeletalinvolved fate; are cells that signals the crest identify to is neural goal Our and ectoderm, endoderm, the The facial skeleton in all vertebrates arises from complex cellular interactions that occur between United States California,RegenerativeMedicine,Southern Celland Los of Stem UniversityCenter for Angeles, F. Stellabotte,G.Crump Jagged-Notch signalingpatternsthefacialskeleton Organogenesis SESSION IV eurd agdNth inln wt te etal rqie Edtei (d1 morphogen. (Edn1) Endothelin required ventrally generated the We with dorsally between signaling relationship the Jagged-Notch study to required wanted we function, Edn1 reduced with skeletons larvae the resembled of embryos NICD of skeletons the Since skeleton. jaw ventral more the of reduction a and expression gene ventral-specific of loss a saw we stages, patterning skeletal during (NICD) domain intracellular Notch the misexpressing by signaling Notch activated we when fact, In phenotype. opposite the show would signaling Notch of activation that predicted jag1b function findthat with a we morpholino (MO) resulted Notch2 of in arches, reduction addition, dorsal-specificIn pharyngeal domain. skeletalventral the defects two in similarpattern complementary toa first in thatexpressed seen the in of domain dorsal Notch the which in expressed examined is next we ligand, signaling Notch a receptor is involved in patterning the is facial skeleton along the dorsoventral Jag1b axis. Whereas Since genes. expressed morphology. We show b1105 Furthermore, we wanted to identify additional DV-specific genes that control skeletal control that DV-specific genes additional identify to wanted we Furthermore, mutants. Since Jagged-Notch inhibition resulted in dorsal-specific skeletal defects, we jagged1b jag1b dlx3b, b1105 , and dlx5a, (jag1b jag1b specifies upper jaw identity by inhibiting the expression of ventrally

and b1105 edn1 ), in ventral skeletal precursors. Recently,precursors. skeletal ventral a in discovered dlx6a we which results in dorsal skeletal elements acquiring a ventral a acquiring elements skeletal dorsal in results which tf216b double mutants in which we find the facial skeleton is skeleton facial the find we which in mutants double 19 is notch2 jag1b 55

Talks Talks levels. Interestingly, levels. markers 56 of formation the during proteins core to addition as such chain proteoglycans, side glycosaminoglycan initiate cartilage and bone development during zebrafish endochondral ossification. Xylosyltransferases in mutation loss-of-function a of effects the reveal we Here, vivo. matrix, extracellular abundant remarkably in little is known about the themselves influence of proteoglycans onimmerse skeletal cell differentiation cells skeletogenic that fact the Despite Institute ofNeuroscience, University ofOregon, Eugene, USA B. F. EamesandC.B.Kimmel Positive andnegative feedback fromproteoglycansduringchondrocyte differentiation /- and anegativeroleduringchondrocytematuration. differentiation chondrocyte overt of stages initial during role positive a play proteoglycans that idtp sbig. t al sae o oet hnrgnss in chondrogenesis overt of the sameas stages early roughly is At siblings. these elements wild-type of patterning the although ossification, endochondral in perichondral bone, even though even bone, perichondral in genes are expressed in expressed are genes ihh maturation, chondrocyte initiate prematurely cartilages mutant that hypothesis a of support In perichondrium. overlying the in precursors osteoblast to signal to known are and chondrocytes perichondrium. Osteoinductive factors, such as Indian hedgehog (Ihh), are expressed in maturing zebrafish demonstrate decreased Alcian blue staining in cartilage elements undergoing incartilage blue staining Alcian decreased demonstrate zebrafish 20 sox9b and col2a1b xylt1 wih s xrse hgl i criae Accordingly, cartilage. in highly expressed is which , aggrecan -/-

xylt1 zebrafish also have increased and premature Alizarin red staining red Alizarin premature and increased have also zebrafish are expressed in similar domains as wild-type, but at decreased at but wild-type, as domains similar in expressed are -/-

cartilage earlier than in wild type. In total, these data suggest data these total, In type. wild in than earlier cartilage xylt1 does not appear to be expressed highly in developing in highly expressed be to appear not does xylt1 xylosyltransferase1 -/-

uat, h cartilage the mutants, Organogenesis SESSION IV ) on (xylt1) xylt1 in - D.Y.R. Stainier these genescontributestothegrowthandsurvivalofcolorectal cancercells. of orthologues human the of Wederegulation formation. the whether determining currently are well with the expression patterns of these genes during the rapid growth phase of digestive organ in our mutants, though unexpected in view of the critical nature of the defective genes, correlate observe we organ-specific phenotypes The cells. epithelial intestinal of survival and growth the regulation. for essential being and as genes cellular important expression of number Together a identified have gene studies our in splicing class minor of role important an consensus indicate genome distinctive with the throughout introns 1%) class minor than of distribution non-random (less and conserved introns The sites. splice of fraction small a processes that complex encoding a component of the U12-dependent minor spliceosome, a highly conserved molecular mutants, cellular essential of variety a in roles processes, play while genes the seventh cloned is a our novel gene. of The growth seven of the of intestinal epithelium out in two six Interestingly,of the 4dpf. at development pancreas and liver intestinal, in defects with mutants 100 approximately n dsutd iooe ignss a rtcl rcs i poieaig el, nldn cancer including cells, proliferating in cells. Lastly, process the positional cloning critical of a biogenesis, ribosome disrupted processing RNA ribosomal and abnormal to lead all genes different three are in mutations cells Here, Apoptotic rare. wildtype. to compared unfolded relatively and disorganized dysplastic, is the on out carried was UCSF.which at screen, screen This .. Trotter A.J. 1 J.K. Heath J.K. intestinal zebrafish disrupt pathways development cellular essential diverse of components in Mutations Organogenesis SESSION IV of Biochemistry and Biophysics, University of California, San Francisco, USA and USAFrancisco, San California, of University Biophysics, and Biochemistry of opooy Mawie h itsia eihlu in epithelium theendosome- intestinal the and Meanwhile size morphology. through both in abnormal are cells proteins epithelial intestinal trinculo in of lysosomes and endosomes trafficking including processes, microscopy,pathway.electron degradative and lysosome confocal Wethat using shown, have cellular numerous to (H+) vacuolar of subunit ATPase(V-ATPase), contributes that pump proton well-characterized a o xlr ter oeta rl i te eeomn ad rgeso o clrca cne. To Liver cancer. the in colorectal identified mutants of intestinal seven progression cloned and positionally have development we aim, the this infulfill role potential their explore to and development intestinal vertebrate in functions with genes zebrafish identify to is aim Our Eliza HallInstituteofMedicalResearch, Royal Parade, Parkville, Australia of each round of mitosis. In mitosis. of round each of encodes a nuclear pore component that is essential for the assembly of nuclear pores at the end premature stop codon in the numbers of intestinal epithelial cells undergo apoptosis. The molecular lesion in Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Australia; flotte lotte 1 1 , A.P.Badrock H Verkade H. , 2

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Talks Talks 58 potentiate properdifferentiationintoliverandpancreas. placing the bipotential hepatopancreatic progenitor cells in the right embryonic environment to cdx genes are responsible for providing the right positional cues along the anterior-posterior axis, cdx1b and of depletion Combined signals. BMP and Acid Retinoic Fgf, to cells progenitor naïve of exposure the influencing environment, signaling and endoderm the in progenitors between talk of depletion that demonstrate we Furthermore fates. pancreas exocrine this gene leads to a two severe expansion these of the endocrine that pancreas fate in suggest detriment anterior-mostpromotes Cdx4 of depletion and zebrafish of in precursors hepatopancreatic in fates both studies liver and previous and foregut embryonic endoderm-derived tissues arise from a subset of common the bipotential progenitors. We found that in cells endoderm from antero- develop during progenitors pancreas critically and Liver tube. function gut the to of development known and patterning posterior are and vertebrates among conserved highly are The Caudal-related (Cdx) genes are homeobox genes, members of expression. gene the of orchestration parahox spatial and cluster.temporal precise a requires Cdx patterning Embryonic genes Institute,Hubrecht Utrecht, The Netherlands A. Faro, J.Korving,H.Begthel,E.deBruijn,V. Gouriev, E.CuppenandH.Clevers progenitor cellpopulationinzebrafish CDX genes promote hepatic fate and repress pancreatic fate within bipotential hepatopancreatic 22 cdx4 revealed that all cdx genes act redundantly during endoderm organogenesis. Thus modifies the cross the modifies cdx4 Organogenesis SESSION IV cdx1a, hc cnrbt frhr ucinl nlss o xmn te oe i te neir chamber anterior the in roles the examine to analysis maintenance. analyses, functional profiling gene further by of tissues set contribute ocular comprehensive which adult provide the we together, in All genes hybridisation. cornea situ the in of distribution spatial mapped We analysis. relative amount of the cornea genes over the skin and confirmed the results from the microarray the adult cornea in comparison to the skin (P<0.01). Further RT-qPCR quantification established in whichisinitially enriched are that genes cornea ~165 presented analysis tissue, Microarray level. ultra-structure the reference asa gene skin analysed at similarities anatomical high the shares we and stages embryonic using during chamber,cornea the with continuous anterior cornea the adult the of of maintenance profile expression the in involved genes identify To the of polarity planar 2) formation; corneal epitheliumwasregulatedbymultiplepathwaysofEGF,endothelium NotchandHh/PKA. corneal the for required are signallings FGF cornea morphology were analysed by time-lapse imaging and electron microscopy: 1) EGF and the on inhibitors chemical of effects the formation, chamber anterior the in involved pathways signalling the into chamber.insights Toanterior gain the contribute that source major a as hpf a reporter earlier discriminate to one allows expression from imaging the later. lapse This time “pulse-chase” imaging cancelled the out to prior photo-conversion transcription activity by using a unique feature established of We EosFP,origin. crest neural a the of photo-convertible structure fluorescencea protein; cornea, the on focused we level, molecular thelens between the at chamber anterior the of development and origin humour the understanding for basis the establish aqueous with Tovision. normal a filled for crucial hence retina, space the on focused light clear the guides which cornea a and is eye the of chamber Anterior ITG, FZK, Karlsruhe, Germany M. Takamiya, B.Weger, U.Straehle Zebrafish anterior chamber development andmaintenance Organogenesis SESSION IV -active cluster of non-ectomesenchymal neural crests in the pharyngeal arch at 18-20 at arch pharyngeal the in crests neural non-ectomesenchymal of cluster sox10-active transgenic line to follow neural crests, but also to visualize to also but crests, neural follow to line transgenic sox10::eosFP sox10-inactive cells to reveal 23 de novo de sox10 59

Talks Talks Lipmann Institute,Lipmann Jena, Germany; 1 R.I. Handin Division, Brigham and Women’s Hospital, Boston, USA; found functional donor-derived GFP circulation of unlabeled wild-type or kidney-injured recipient fish. Following transplantation we tagged with a kidney-specific green fluorescence protein (GFP) transgene were injected into the cell population in adult fish, we carriedstem out transplantation experiments.renal Donor cellspresumptive genetically the characterize Tofurther nephrons. existing with fuse basophilic that structures into coalesce and expand to appear aggregates cells that express These the marrow.transcription factor kidney the within cells factor transcription renal the of expression increased showed kidneys fish adult injury.damage, to kidney response gentamicin-induced in Following kidney) the of unit functional (the nephrons new regenerating in involved is that kidney zebrafish the in population cell stem renal a of existence the supporting evidence kidney.present the we Here, Zebrafish possess tremendous regenerative capacities and can regrow a range of organs, including Biochemistry, SchoolofMedicine, University ofPittsburgh Pittsburgh, USA 60 C.Q. Diep Renal stemcellsgive risetonewnephronsduringzebrafish kidneyregeneration From stem renal cells the Single expressing of development. larval origin during kidney adult the the of determine formation Tothe examined nephrons. we cells, new form and circulation from the to kidney back home can that population cell stem/progenitor renal a possesses kidney zebrafish adult the that suggest findings These fish. recipient wild-type of 9% in and regeneration, kidney source fordenovo nephronformationinresponsetorenalinjury. from embryonic kidney. Taken together, we propose that the adult zebrafish kidney is initially formed of expression initiate expand, to USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, USA; 24 lhx1a-expressing renal stem cells, and these cells persist in the kidney marrow to serve as a timecourse analyses of double transgenic larvae, these larvae, transgenic double of analyses timecourse vivo in 5 rerc-cilrUiest, ea Germany; Jena,Friedrich-Schiller-University, 1 , D. Ma lhx1a appeared just posterior to the swim bladder at around 10 days post-fertilization. 2 , A.J. Davidson 2 ,G. Djordjevic 1 , and differentiate into adult nephrons that fuse with the with fuse that nephrons adult into differentiate and wt1b, 1 4 , F. Bollig Section on Model Synaptic Systems,Synaptic Model NIH/NIAAA,on Section Bethesda, + nephrons in 47% of the recipient fish that were undergoing 3 , T. Ikenaga wt1b and then epithelialize into primitive nephron 6 eatet f oeua Gntc and Genetics Molecular of Department 3 4 Leibniz; Institute for Age Research, Fritz , F. Ono in single mesenchymal-like single in lhx1a 4 , C. Englert lhx1a + 5 cells were found were cells , N.A. Hukriede Organogenesis SESSION IV 2 Hematology 6 , 1960. Laser-induced Site-Specific gene Temporally-Controlledexpression in (2009); specific cells of M transgenicM,MC,WarrenSato-Maeda 2.JT, Halloran zebrafish.F, Brand Su PH,KuwadaJY,Z,Lele Krone Development (2000);W D, Shoji 127(9):1953- Freudenreich J, Recombination inZebrafish. PLoS ONE4(2): Kaslin e4640. S, Hans 1. the rapidzebrafishdevelopmentthatismuchfasterthandegradationofEGFP. of advantage takes it because embryo zebrafish early the in studies mapping fate facilitate will certain structure is formed by the early Cre expressing cells. Hence, the conditional a final reporter that line reveal its might and reached labeled be not will lineage the has of portion domain a Consequently,only expansion. expression Cre the when stage certain a at activated be line can reporter conditional the domain, expression Cre a of lineage entire the labels always that reporter constitutive a to contrast in Furthermore, examined. stages all at mapping fate genetic that this conditional red-to-green reporter line temperature-inducible zebrafish promoter which has been the shown to express towards in a strong and turned ubiquitous fashion [2]. thus We will we show line reporter red-to-green gastrulation and mid-somitogenesis stages which limits the use of this line. To establish a faithful showed that Cre is in fact highly efficient in this organism. Furthermore, temporal control of control temporal Furthermore, organism. this in recombination canbeachievedbyusingtheligand-inducibleCreER efficient werecently highly fact in transgenesis, is mediated Cre transposon that butusing showed low very previously was efficiency recombination Cre-mediated zebrafish, In organisms. model vertebrate other in absent almost but mouse, in technology powerful a is Cre recombinase site-specific the of use Conventional Biotec/CRTD, TU-Dresden, Dresden, Germany S. Hans,J.Kaslin,D.Freudenreich,M.Brand DsRed2-loxP-EGFP) reporter red-to-green conditional the using tracing lineage genetic Improved Emerging technologies SESSION V of DsRed2 for up to 7 days, 7 to up for DsRed2 expression of strong see we although However, event. recombination successful a after EGFP to Xenopus Elongation Factor 1 alpha 1 Factor Elongation line Xenopus reporter red-to-green a generated previously we Tg(EF1α: mapping loxP-DsRed2-loxP-EGFP fate ). This genetic line expresses allow DsRed2 under to the control order of the In ubiquitous in situ in ) promoter in the absence of Cre activity, but changes activity,but Cre of absence the in promoter (EF1α) analysis reveals strong ubiquitous expression only during only expression ubiquitous strong reveals analysis Tg(hsp70: loxP-DsRed2-loxP-EGFP) indeed allows T2 25 [1]. : loxP- Tg(hsp70: hsp70 61

Talks Talks between the different cell systems used in our study, illustrating the relevance of performing of relevance the illustrating study, single-molecule microscopyinvivo . our in used systems cell different the between significantly differs organization membrane This diffusion proteins. membrane of diffusion the of confining type the in differ which exist, behavior they display. anchor Furthermore, membrane we demonstrate two the YFP-tagged occurrence that of the membrane demonstrate microdomains and of cells populations mammalian in findings previous with line in are experiments our of results The approach. this for used was setup microscopy (TIRF) reflection internal total A protein. tagged fluorescently the encoding RNA with injected previously embryos, zebrafish two-day-old of skin the in cells epidermal in performed was organisms living in imaging Third, cells. primary dissociated the using performed was Subsequently,imaging stage. sphere the at wereinjected embryos zebrafish RNAwith Second, dissociated were embryos injected the and protein tagged fluorescently the encoding setup. microscopy fluorescence wide-field a using three-step approach was used. First, individual a molecules this, were achieve visualized in to cultured order (ZF4) In cells membrane. plasma the in anchored proteins for model a as serve to shown been anchor, has membrane which H-Ras human the to fused (YFP) protein fluorescent molecule microscopy to the level of a living organism. As a molecule of interest we bya used yellow and detection cells in culture. In the present study, the zebrafish was used as a model system to extend single- proteins tagged fluorescently of excitation sensitive CCDcamera.However, untilnowthistechnologyhasonlybeenappliedtoindividual laser using years, several for Studying the dynamics of individual proteins by single-molecule microscopy has been available of LifeProcesses, Institute ofPhysics (LION), Leiden University, The Netherlands and H.P. Spaink 62 M. J.M. Schaaf The zebrafish asamodelsystemforinvivo single-moleculemicroscopy 1 Molecular Cell Biology, Institute of Biology (IBL), Leiden University,Leiden(IBL), Biology, CellBiology Netherlands;Molecular of The Institute 26 1 , W. J.A. Koopmans 1 2 , T. Meckel 2 , J. van Noort 2 , B. E. Snaar-Jagalska Emerging technologies 1 SESSION V , T. S. Schmidt 2 Physics 2

integration site. the investigating are and phenotypes mutant possibility to use I-SceI meganuclease for generation of local deletions involving the of transposon reversion for recombinase Cre of expression To complement this insertional mutagenesis approach, we are lines. generating lines with tissue-specific trap gene the of each for identified been have genes affected Candidate blood. and vasculature pancreas, system, nervous the as such specific, highly to mosaic weak to ubiquitous bright very 12 isolated and transposon this with injected embryos from vector. raised trap fish Togene 70 reporting screened we date, self- a levels maybe UAS:eGFP,making added thus also we expression ToGal4-VP16 very low Gal4-VP16. detect by at reporter mRFP the replace to us prompted genes expressed considerations other and into This detect. to impossible integrations is that reporter fluorescent a of termination/ drawback potential One events. trapping gene mRFPfor used reporter vectors the initial as Our and transcriptional for integrationintogenesandefficientdisruptionofgeneexpression. acceptor selection stringent splice enables reporter.advances the two from these removed of was Combination codon initiation translation fish-derived the specificity, Toincrease used. were sequences mutagenicity,polyadenylation transposons. breaking gene increase of specificity and To mutagenicity achieve to made were advances gene from built Twointegration. key upon are expression gene disrupt vectors to known components Our transposon breaking zebrafish. the in pattern) expression and phenotype (mutant characteristics two these address simultaneously mutantto tools transposon-based the developing are We nor pattern expression the neither Thus, phenotype isrevealeduntilatransgenicmousegeneratedfromtheseEScells. cells. ES in out carried are experiments these system model a as mouse of limitations to Due traps. gene using mutagenesis insertional gene, affected genome-wide undertaking are consortia several the mouse, the In non-mutagenic. inherently is of but pattern expression the reveals It trap. enhancer is approach simultaneously used widely that and approaches successful most the experimental Drosophila, In system. model develop any pattern. in characteristics two expression to these address and challenging phenotype extremely function been of has loss It are gene any of characteristics key Two ofBiology,Department Temple University, Philadelphia, USA D. Balciunas,D.Nagelberg, K.Gonzalez,J.Balciuniene Transposon-based insertionalmutagenesisplatform Emerging technologies SESSION V gene trap lines. Gene trap expression patterns range from range patterns expression trap Gene lines. trap gene fide bona 27 63

Talks Talks 2 1 64 R. W. Köster Optimized Gal4geneticsforpermanentgeneexpressionmappinginzebrafish and maintained expression by Kalooping, a technique that can be used for permanent spatio- temporal geneticfatemappingandtargeted transgeneexpressioninzebrafish. permanent for used be can that technique a Kalooping, by expression maintained and cells in rhombomere 5 during stages of early embryogenesis. These data demonstrate prolonged that the secondary octaval nucleus in the adult hindbrain is likely derived from mediated autoregulatory mechanism to live adult structures. We demonstrate its use by showing KalTA4-a KalTA4via transient embryogenesis relates during which expression “Kaloop”, strain rendered KalTA4 be activationindetail.Thesestudiesleadtothedevelopmentofaself-maintainingeffector can and transgenes tissue-specific of kinetics transactivation the characterized have UAS-dependent we Furthermore, homozygous. transactivate reliably activator strains KalTA4 fluorescent These red strains. self-reporting of the collection Witha provide expression. we expression, transgene system of effects dosage studying a of for help allow on to dependent zebrafish.In numbers activation usein copy transgene KalTA4-mediatedUAS for in vivo data in genetics quantitative for Gal4-UAS offer we of (KalTA4) addition, optimization Gal4-activator systematic an optimized a establishing report zebrafish we Here introduced. been have . Drosophila Recently this system was adapted for zebrafish research and promising applications in studies genetic sophisticated for possibilities the expanded immensely has system Gal4-UAS combinatorial the of development The control. specific tissue and temporal with study function gene to technology powerful a is activation transgene conditional for genetics Combinatorial Germany uwgMxmlasUiest Mnc, eatet f ilg I, Planegg-Martinsried, II, Biology of Department Munich, Germany; Neuherberg, Ludwig-Maximilians-University Genetics, Developmental of Institute München, Zentrum Helmholtz 28 rnpsnmdae KlA ehne ta sre bae fr eta nervous central for biased screen trap enhancer KalTA4 transposon-mediated Tol2 1 , M.Distel 1 , M.F. Wullimann 2 , Emerging technologies egr2b-expressing SESSION V and illustratethepotentialofthesephotoactivatablereagentsinfunctionalgenomicsresearch. cMOs targeting develop to them applied successfully and design cMO for principles basic established have we structure, cMO and between adhesion, cell relationship the investigated and procedure synthetic cMO our optimized also have we pathways, factors, signaling developmental with extracellular matrix production. To associated facilitate the proteins application of this including approach to other transcription have 70 we strategy, this approximately Using analysis. discovered expression gene microarray and sorting, cell activated fluorescence- fluorophores, photoactivatable cMOs, combining by manner tissue-specific a in whole-embryo analyses of the transcriptional targets of the discover T-boxto cMOs of applicationgene the here report we and organisms, transparent optically other and zebrafish in manipulations experimental such enable Caged oligonucleotides (cMO) precision. morpholino temporal and spatial with programs genetic these alter to ability an requires physiology and development vertebrate regulate that mechanisms molecular the Deciphering Chemical andSystems Biology, University SchoolofMedicine, Stanford Stanford, USA J. K.Chen,IA.Shestopalov, X.Ouyang Transcriptional targetdiscovery throughcagedmorpholinotechnologies Emerging technologies SESSION V other patterning genes. Our findings establish the generality of cMO technologies thermodynamics, and thermodynamics, vitro in ntl mutant/morphant transcriptome, we have identified dpnet ee ta cnrbt t ntcod formation, notochord to contribute that genes ntl-dependent no ( tail ntl). In contrast to previous studies that conducted activity. Through these studies, these Through activity.vivo in 29 ntl targets 65

Talks Talks duplications. enable detection of intra-chromosomal events including chromosomal inversions, deletions and a multi- using chromosome, color labeling approach, specific will eventually lead to a a multi-color chromosome probe set that would within BACs chromosome-specific individual label to the rapid identification of suspected chromosomal translocations in mutant strains. Future efforts chromosomal translocation using two-color FISH. The establishment of this resource will enable intervals. Additionally,Mb 1 approximately at chosen chromosomes a identified also have we zebrafish each for (BACs) chromosomes artificial bacterial chromosome-specific of pools using probes paint chromosome-specific developed have We chromosomes. marker supernumerary ambiguous and inter-chromosomal rearrangements of identification rapid permits probes DNA DNAprobes that of “coat” combinations entire chromosomes containing with probes a given painting fluorescentchromosome color.developed now Thehave development ofwe these gap, can they which at scale identify chromosomal the rearrangements, amplificationsbut andrearrangements, translocations ischromosomal limited.gross identifying To bridgefor this useful andnear- be to for proven near-telomeric has which 2007) al., et probes (Freeman chromosomes zebrafish all (FISH) of regions centromeric hybridization situ in Recently, fluorescence lacking. still developed is we alterations chromosomal specific visualize and mutants of genomes the characterize to probes cytogenetic molecular of cancer,establishment and the biology cell development, vertebrate studying for organism model a as zebrafish of importance the Despite 66 K. H.Brown rearrangements andmarkerchromosomes inter-chromosomal of identification the for chromosomes zebrafish of painting Chromosome USA; 1 Department Department of Pathology, Brigham and Women’s Hospital and Harvard Medical School, Boston, 30 2 School ofHealthSciences, Purdue University, West Lafayette, USA 1 , J.L.Freeman 2 ,C.Lee 1 Emerging technologies SESSION V J.C. Smith 3 F.C.Wardle A mesodermalgeneregulatorynetwork directedby T-domain transcription factors Genomic workshop SESSION VI early zebrafishembryo. describe a preliminary Gene Regulatory Network for mesoderm formation and patterning in the we data these using and T-domainfactors, other of targets similar identify to Using begun studies. have we vitro methods in by identified previously (TBS) site binding T-box consensus the network of other transcription factors and we identify an in vivo Ntla binding site that resembles massively that and plays a central role microarrays in mesoderm formation. We genomic have found that with Ntla parallel sequencing regulates we have identified targetsa of combined Nodownstream tail a (Ntla), a zebrafish Brachyury ortholog immunoprecipitation chromatin Using Bioinformatics Institute, Cambridge, UK 1 et Pyilg, eeomn & ersine Uiest o Cmrde UK; Cambridge, of University Neuroscience, & Development Physiology, Dept. National Institute for Medical Research, London, UK; 2 3 R.. Morley R..H. , 1 K Lachani K. , 2 .. Nelson A.C. , 2 Gurdon Inst., University of Cambridge,UK; 3 D Keefe D. , 4 MJ Gilchrist M.J. , 31 2 P Flicek P. , 4 European European 67 4 ,

Talks Talks signalling pathwayshavebeenperturbed. which in embryos in phenotypes expression gene detecting in tool analysis image our of utility screens. toxicological or pharmaceutical genetic, for embryos high throughput analysis of spatial distribution of a variety of fluorescence readouts in zebrafish the for suitable also is here described technology The traps. enhancer as such studies reporter promoter sequence in cis-regulatory interactions and provide a valuable promoter resource core for transgenic the of promoter importance enhancer-core the 203 underscore specificities between interaction These specificities combinations. pipeline interaction imaging of embryo diversity the a Using demonstrate shape. we reference dimensional two virtual a onto images Domain specificity of Venus reporter expression was evaluated by warping of individual embryo allowed the registration of reporter activity in Automated intelligent almost microscopy and 18 custom built thousand embryo detection and microinjected segmentation software zebrafish throughput embryos. high aim of mapping a the of the with application embryos zebrafish in expression and reporter of specificity domain development registering for pipeline the report we Here the by embryo. provided potential transparent the exploit to technologies imaging of automation for need is there complexity of the vertebrate animal for a variety of phenotype screening applications. However, Zebrafish embryos offer a unique combination of high throughput capabilities and the organismal Forschungszentrum Karlsruhe, EggensteinLeopoldshafen, Germany U. Liebel 1 68 J. Gehrig Automated highthroughputmappingofpromoter-enhancer interactions inzebrafish embryos Leopoldshafen,Germany; n Dna Sine, nvriy f imnhm UK; Birmingham, of University Sciences, Dental and Institute of Toxicology andGenetics, Forschungszentrum Karlsruhe, Eggenstein- 32 1,2 1 andF. Mueller , M. Reischl interaction specificities between cis-regulatory modules and core promoters. 3 , E. Kalmar 1,2 2 Department of Medical and Molecular Genetics, College of Medical Genetics,of CollegeMolecular and Medical of Department 1 , Y. Hadzhiev 1,2 , M. Ferg 3 nttt o ple Cmue Science, Computer Applied of Institute 1 , A. Zaucker To this end we are exploring the exploring are Towe end this 1,2 , C. Song Genomic workshop SESSION VI 1 , S. Schindler 1 , 1 S. M.Burgess Identifying thegenenetwork involved ininnerearcellregeneration intheadultzebrafish Genomic workshop SESSION VI vns eg cl dvso ad ifrnito) uig ar el eeeain o e a better a get to regeneration cell hair during differentiation) and understanding ofthefunctionsthosegenes/pathwaysinregenerationprocess. division cell (e.g. cellular various with events genes/pathways candidate new the associating on working now are We candidate pathwaysinvolvedinthehaircellregeneration. new as well as genes candidate new suggested data TagProfiling the of analysis Our and/ analysis. developed pathway and have Weanalysis, ontological point. assignment, gene time for tools each bioinformatic at various utilized expression or gene of view deep very a pool, us mRNA the giving from “tags” gene more or million 3 generates technique the nature, in SAGE manner. to high-throughput Similar and in-depth an in profiles expression gene the generate to the various time points. Here we used a new technique, Tag Profiling (Digital Gene Expression), at expression in during increase significant with genes those points on particularly focusing regeneration, time the regeneration different at the subsequent profiles expression gene inner-ear characterize the and determined We zebrafish process. adult the of epithelium saccular the in loss cell hair induce to us enabled which protocol, noise-exposure previous a Wemodified for geneexpressionprofiling. response used for hair cell regeneration in zebrafish using next generation newones inner sequencingfor ear techniques hair with cell regeneration in requiref adult vertebrates. As network an initial step, cells we identifiedgene the transcriptional the hair understand to is goal in ultimate lost Our deficiencies adulthood. replace throughout permanent can various zebrafish in contrast, resulting In replaced, sensation. neuroepithelia. not vestibuloauditory the are in cells units hair mechanotransductory lost the mammals, are In ear inner the of cells hair Sensory Park, USA National Human Genome Research Institute,ResearchBethesda, USA;Genome Human National 1 , J.Liang 1 , G.Renaud 1 , T. Wolfsberg 1 , and A. N.Popper 2 University of Maryland, CollegeMaryland, of University 33 2 69

Talks Talks indels and other structural variants in the Wildtype strain of Zebrafish will be presented and presented be will Zebrafish of discussed. strain Wildtype the in variants structural other and indels small polymorphism, coverage single-nucleotide regarding 25Xsequence Updates genome. reference inaverage Zebrafish the of resulting genome, reference Zebrafish Sanger onto reads analyzer.Genome 11Illumina from 110the data Weon short sequence runs the of aligned GB strain-to-reference comparison for genome-wide sequence variant discovery. We collected that over strains Zebrafish facilitate will This India. in bodies water Wildtype the a from directly caught Zebrafish of Strain grown lab these on in captured sequenced have we variations, was focused genomic Toto studies. related genome questions diversity initial address genetic been true no has or little clones; work genetic represent genomics functionally main the parallel date, Till massively allows method mapped backtothereferencegenome. This Analyzer. then are which pairs, base Genome 75 to 36 from ranging fragments genomic of millions of Illumina sequencing of technology (MPSS) Sequencing Signature Parallel Massive the using Zebrafish of Strain Wildtype a of genome the Wevariations. sequence genome-wide identify to reference a to compared are sequenced have the for reads short sequence whereby technologies, reference re-sequencing fast of a application allows permits of genome Zebrafish and existence The genetics processes. understanding biological for important of valuable discoveries is sequences DNA by encoded Information V. Scaria Malhotra S. 1 70 Sivasubbu S. Whole-genome sequencingofa Wildtype Strain ofZebrafish for variant discovery Institute ofGenomicsandIntegrative Biology, Delhi, India; 34 1

1 , J. Maini J. , 1 , A. Patowary A. , 1 , M. K.Lalwani M. , 1 , R. Purkanti R. , 1 , N. Singh N. , 1 , M. Swarnkar M. , 1 , A. Singh A. , 1 , A. Ramcharan Singh Ramcharan A. , 1 2 , V.Pandey , Mayo Clinic, Rochester, USA. 1 , S. C. Ekker C. S. , Genomic workshop 1 SESSION VI , R. Chauhan R. , 2 , E. Klee E. , 2 1 , , but byextensivegeneticmappingdata. be the most complete assembly and that it is confirmed not just by sequence and clone overlaps Thus we anticipate that the next release, Zv9, will be very densely covered with genetic markers, to improvethegenomecoverage. one whole genome shotgun read. We have about 40Mb of such sequence, and hope to use this least at by supported is which of much Zv8, current the in not is that sequence identify to able assembly failures. From our 100s of millions of reads - generated without cloning steps - we are assembled well as such by difficulties technical covered or to regions TAcentromeric long and e.g. telomeric due sequences stretches, not certain of unclonability often genome, are human the that in regions even true are is This there sequence. assembly genome any in SNPs Finally 200,000 over selecting currently are we Tuebingenpolymorphic betweenthetwoDHfishforhighresolutionmappingonacustomSNP array. data one sequence and our ~0.1cM AB give Using would each (~65kb). which one panel resolution F2 have 500 a which (currently assembling are F1s we female which Tuebingen of from and haploid set chromosome, doubled a generated a have to we male sequencing) DHAB sequenced the crossing By very a with sequence. genome the researchersto cross mapping mutant a from provide transition rapid and map genetic dense would andthento this chip, Furthermore, length. it’s single along ontoa assembly genome 7.5kb) every SNP (1 the anchoring genetically - clone BAC a of that than finer SNPs resolution a at these map genetically 200,000 over fit to possible is It inversions basedonreadpairmappinganomalies. on changes in read sequences, and larger indels (including copy number variantss - CNVs), and identifying SNPs we can also identify small insertions and deletions (indels) in nucleotide the genome based single million 10 polymorphisms (SNPs) between over the DHAB fish and identified Tuebingen reference genome.we In addition to doing so By sequence. genome reference the to reads these mapped and fish, AB male (DH) have haploid doubled we a coverage sequencing 25x to generation sequenced next Illumina Using strains. lab common two these even variation between genetic the of knowledge limited have we YetAB. especially strains, laboratories other Many use strain. Tübingen the on based is project sequencing genome zebrafish The of Oregon, Eugene, USA 1 M. Clark Genetic diversity between Tübingen and AB strains, andanewultra-dense geneticmap Genomic workshop SESSION VI Vertebrate Development and Genetics, WTSI, Hinxton, UK; 1 ,C.Torroja 1 , J.Postlethwaite 2 , D.Stemple 1 2 Institute of Neuroscience, University 35 71

Talks Talks 1 72 Spaink H. High throughputscreeningofinnateimmuneresponsesinzebrafish andcarpembryos In recent years the zebrafish has been shown to be an excellent model for studyingthe for model be anexcellent to responses shown towards pathogens such as been has zebrafish mechanisms of the the innate immune defense years against pathogens. We recent have shown that In transcriptome Jagalska which canbefurtheranalyzedintransgeniczebrafishmodels. show that it is now possible to undertake chemical compound screens in carp fish the results of the hallmarks of innate immune responses in both fish species are extremely similar. Our results a obtained that showing zebrafish with have obtained results compare Weto order in carp the screening. of sequence shotgun throughput high for embryos of quantities limiting overcome common the instance For assays. carp that throughput can yield hundreds of thousands of embryos for each high fish after in vitro fertilization can for zebrafish the of relatives close employ also to approach this in profitable highly be can it Wethat sorting. show embryo with coupled profiling scanning laser flow-through using symptoms disease screening for Biometrica) (Union can also be extended to studies a such high that throughput show level. will we For presentation this this we In have assays. applied xeno-transplantation the using Copas instance XL biosorter only relevant to infectious diseases but also to the study of immune responses not to are cancer results cells, These system. for immune innate the of players key of functions the in insights new deep transcriptomic of sequencing, combinations Using morpholino knockdown systems. and mammalian transgenic in reporter fishresponses technologiesto we havesimilar obtained very Leiden University, Leiden, The Netherlands; 36 1 , A. Meijer 1 R Dirks R. , 1 2 R Carvalho R. , 2 O Stockhammer O. , Mycobacterium marinum and 2 ZF-screens B.V. Leiden, The Netherlands 1,2 S He S. , 1 A Zakrzewska A. , Salmonella typhimurium are Genomic workshop SESSION VI 1 E Snaar- E. , and journals. completes a new and important data feedback among authors, electronic data systems like ZFIN, with journals and biocurators to facilitate accurate data exchange and reporting. option. publication open-access an provide now journals Many journal. a choosing when considered be should and databases public in data of accessibility the limit issues These available. readily not do journals Some give permission important. to reproduce also figures in publicis databases, and theformat articles themselvesand may not be journal of choice The publication. before not eliminate all ambiguity. As a reviewer, one can identify ambiguities and missing information morpholino? Which developmental specific! stage? References to prior publications are helpful, but may be - enemy the genes. new is naming Ambiguity before committees nomenclature consult and nomenclature, mutant and gene current for databases organism model check possible, whenever sequences and numbers most the extract completely.and provide Namely,correctly should represented authors are data their and publications read carefully captured salient data for who entry into a database. Authors can facilitate the curation process and ensure that be professionals publication. trained can each in are that available information Biocurators information the of of precision the from type upon largely dependent and come critically are data amount those quality, and data, the their However, as savingtime publications. good bench, as the only to are Databases going resources. before and hypotheses community. refine research wider researchers the help on databases work your Public of impact the amplifying data, other with data your link and integrate (zfin.org), Network Information Zebrafish the as such databases, Public preliminary stepinalarger processthatinvolvespublicdatabases. data. communicating in step final the once was manuscript a Publishing Zebrafish Information Network, University ofOregon, Eugene, USA M.A. HaendelandZFINstaff data stream Amplify the impact of your research: Ensure that your data can be integrated into the electronic Genomic workshop SESSION VI This is the friendliest format for a public database. Authors can help by working together hc “h” ee Which gene? “shh” Which 36 bis lee Which allele? fgf3 Now, publication is a Now,is publication This interaction accession pax2a 73

Talks Talks 74 efficient modelsystemforvalidatinganti-cancereffectsof mitotic kinaseinhibitors. an as serve will embryos zebrafish and kinases, mitotic and mitosis of mechanisms molecular the studying in Taken tool live-imaging. valuable with a is together,zebrafish that conclude we (SAC). Moreover, we were able to monitor the effects of Aurora A inhibitors CIN at in the cancer.single cell As level a result, loss of Plk1 resulted in the activation of spindle assemblyc checkpoint micro nuclei, and defects in centrosome numbers and maturation, all of which are hallmarks of bridges, chromosome pairs, chromosome sister unsegregated accompanied arrest mitotic The Plk1 of loss the that found resulted in we a delay in mitosis developed, for hours while normal we zebrafish embryos dividefish in 12-17 minutes. transgenic H2B-GFP a in imaging live-cell Using organism. living a in cells single analyze to ability the is zebrafish to advantage great A with somedistinctpatternscomparedthoseofPlk1morphants. results similar displayed AAurora of knockdown Morpholino embryos. developing in integrity genetic the in crucial is and stages multiple in mitosis through progression for essential is Plk1 that indicate results These aneuploidy. exhibiting defects, mitotic noted the with divisions death more before two or one underwent cells embryogenic zebrafish Interestingly, attachments. kinetochore-microtubule unstable and numbers, centrosome aberrant outgrowth, microtubule morpholinos resulted in growth defects followed by apoptosis. Loss of Plk1 resulted in defective microtubule using Aurora A or and Plk1 of loss maturation embryogenesis, zebrafish centrosome In roles. distinct in have they outgrowth, implicated both are A Aurora and Plk1 While A. kinases as anti-cancer drugs. We mitotic of inhibitors have of efficacy the validate to adopted specifically be can zebrafish that data this focusedon based on propose twoWe mitosis. studying mitotic for system kinases, valuable a Plk1 is zebrafish andthat showing Aurora data present We alternative invivo system. in cell cycle has control genes, development of inhibitors development of mitotic kinases requires validation drug in an anti-cancer Recently, cancer. focused on CIN and the mitotic kinases. Because established cancer cell lines harbor mutations understanding to central is but biology basic aberrant in question fundamental a byonly not is mitosis of characterized regulation the understanding Therefore, (CIN), instability Chromosome chromosome structures exhibit and numbers. Unregulated mitosis is implicated in aneuploidy and CIN. often cancers Aggressive University, National Seoul Sciences, Natural of College Korea Sciences, Biological of Department H. Lee,K-H.Jeong,H-Y.mitosis andchromosome Jeong,J-Y.of Jeong,H-O.Lee,E.Choi theregulation study to instability system model an efficient as Zebrafish 37 SESSION VII Cancer tumorigenesis, andwillfosterthedevelopmentofimproved, targeted therapyofhumanGCTs. Tumors. cell germ understanding for model zebrafish the of relevance the confirm results These BMP signaling activity, whereas signaling is maintained in the differentiated subtype of Yolk Sac made from our zebrafish model, we find that undifferentiated GCTs such as dysgerminomas lack clinically-annotated human 40 of GCTs series a in pathway of signaling BMP the diverse of activity examined histologic also have subtypes. line. We zebrafish In agreement with the predictions and differentiation and to determine the precise mechanisms of tumor development in the We mutant are conducting experiments to further characterize the role of Alk6b in germ GCTs,cell aswellevidenceofalterationsintheexpressionlevelBMPproliferation target genes. germ cell differentiation, we find evidence of impaired BMP signal transduction in the zebrafish hasbeen pathway controlling in Alk6b for role critical a with agreement BMPsignaling In cancers. human various the in implicated of Misregulation gonadogenesis. and germ specification development, cell embryonic differentiation, proliferation, cell of regulation including roles diverse has signaling BMP receptors. of superfamily TGF-beta/BMP the of member a is Alk6b like Kinase6b)inthemutantanimals. termination codon in the type IB Bone Morphogenetic Protein Receptor, Alk6b (Activin narrow Receptor- the critical interval to a 0.6 cM interval containing three genes. We identifiedto azebrafish premature chromosome 10. We carried out further high-resolution recombinational mapping to dominantly is inherited. Wemutation the used interval that haplotype demonstrated analysis of analysis 14 affected Pedigree individuals age. to localize of the months mutation 9 to 7 around of undifferentiated spermatogonia by 4 months of age while heterozygous males develop tumors genetic a forward during screen to GCT identify of cancer susceptibility incidence loci. a high Homozygous adult with males line develop tumors mutant consisting zebrafish a identified We animal modelsofGCT, whichposesafurtherobstacletoourunderstandingofthedisease. no are there Currently damage. kidney and infertility loss, hearing including effects side severe has remained static since the introduction 30 years ago GCTsof treatment of the reasons, cisplatin these For which, therapies. new while of development effective, the impeding known, causes in increasing are The molecular and origins of GCTs and adults the pathways that treatment. determine to response tumor and young cell potential malignant differentiation their are in not vary which histologies, undifferentiated and children infants, affect incidence worldwide. GCTs (GCTs) arise from pluripotent germ cells and can tumors exhibit differentiated and cell Germ Medical Southwestern Biology, Molecular and Center, Medicine Dallas, USA Internal Pediatrics, of Departments J. C.Neumann,G.L.Chandler, andJ.F. Amatruda A Mutationin Alk6b CausesGermCell Tumors inZebrafish Cancer SESSION VII 38 75

Talks Talks differences arecurrentlybeingidentified. salient these cited, technologies the Using phenotypes. severe more the for responsible are that differences genomic and epigenetic genetic, reveal should initial clones T-ALL and final the of acquired, or already present in a subclone now evident through selective pressure. Comparison newly either mutations, somatic from results situations both in seen disease aggressive the that hypothesize We death. to progression rapid with relapse typically but treatment, irradiation to into cells addition, affected T zebrafish from each line malignant show a marked of reduction in tumor burden transplantation inirradiated response recipients leads to a serial more aggressive disease with accelerated progression to death. genes, In cancer-modulating regarding Finally, loci, andcurrentprogresswillbepresented. changes in DNA methylation identify to patterns. MeDIP and loci, These deleted or amplified studies identify to aCGH have genes, dysregulated identify uncovered several combination candidate to oncogenic a analysis transcriptome using micro-array following: are the including technologies we moderate complementary of genes, these the identify To that suppressors. tumor in or hypothesize occurring hits” proto-oncogenes “second we somatic for requirement a genes, represents line cancer-causingeach in mapping T-ALL category,incidence small second relatively the using Regarding identified rapidly were populations. Thesearch forcandidategenesisongoing. regions high-throughput Linked and markers sequencing. (RAD) DNA associated Solexa site restriction using line each in scans a predisposition to develop disease. To identify the genetic lesions, we carried out genome-wide T-ALL penetrance in the lines is incomplete, indicating that the mutated in allthreecategories. gene in each case confers discovery gene for resources valuable are mutants new severity.Our disease modulating those three into overlapping grouped categories: 1) those predisposing to cancer; 2) those directly be causing cancer; and 3) can genes T-ALL Conceptually, (T-ALL). leukemia lymphoblastic acute for therapies contemporary performed a phenotypic mutagenesis screen and identified three new zebrafish models of T cell isrevolutionizing cancer,we cell T of pathogenesis the genetics in involved genes cancer new Toidentify malignancies. human of field evolving rapidly The Departments of Cancer Institute, University ofUtah, SaltLake City, USA; Nipper 76 Meeker N. New Zebrafish Modelsof TCellCancer:ImportantResources forGeneDiscovery 39 5 , S.L.Perkins 1,2 JK Frazer J.K. , 1 Pediatrics, 4 , andN.S.Trede 1,3 2 L Rudner L. , Internal Medicine, 1,3 3 DF Bradley D.F. ,

3 Oncological Sciences, and 1,3 5 ACH Smith A.C.H. , Floragenex Inc., Eugene, USA 1,3 4 Pathology, Huntsman W Horsley W. , SESSION VII Cancer 3 R.W. , interval that cooperates with cooperates that interval Our screen identified genes in the 1q21 region that are recurrently amplified and overexpressed in human melanoma. all examined melanoma. We human in amplification genome of regions in genes candidate of the with cassette the harbors that miniCoopR called vector plasmid a Wedeveloped rate. melanocytes and subsequent melanoma. Tumors arise at 3 to 4 months of age at a reproducible C.C. Ceol Injection of a rescuing melanocyte-deficient Identification of the SETDB1 histone methyltransferase as a new oncogene inmelanoma oncogene anew as Zon methyltransferase Z. L. histone SETDB1 the of Identification Cancer SESSION VII a loss-of-function allele of allele loss-of-function a This approach utilized a zebrafish strain that reliably forms melanomas due to expression of expression to due melanomas forms BRAF reliably that strain zebrafish a utilized approach This progression. melanoma on effect their for strategy intervals these novel within a genes of developed numbers large test have to we Here, defined. molecularly been not have melanoma in number gain at specific genomic loci, and yet most of the genes in these intervals that participate modulation. such of mechanism major a is number development, coupled with invasion and metastasis. In a variety of solid tumors, genomic copy Cancer is initiated and maintained by successive genetic alterations that lead to enhanced tumor Farber CancerInstitute, Boston, USA 1 melanoma lines with amplified scalable andoffersauniqueapproachtodiscoveringoncogenes inhumancancer. robust, is system zebrafish vertebrate animal whole this using oncogenes candidate screening that demonstrates study This proliferation. cell decreases cells melanoma human in SETDB1 of is the major H3K9 histone methyltransferase for tri-methylation in melanoma cells. Knockdown but lines with the lowest level of eaooyOclg, HICide’ Hsia Bso, USA; Boston, Hospital HHMI/Children’s Hematology/Oncology, V600E , an oncogenic variant found in roughly half of all human melanomas, coupled with coupled melanomas, human all of half roughly in found variant oncogenic an , 1 1 J.J. Jane-ValbuenaJ.J. , , Y.H. Houvras promoter driving cDNAs of interest. This vector allows for the testing the for allows vector This interest. of cDNAs driving promoter mitfa SETDB1, an H3K9 histone methyltransferase, as the sole gene in the 1q21.3 mitfa mutant background, melanoma formation is completely suppressed. mitfa minigene into 1 p53. 2 BRAF , A.U. Uong A.U. , SETDB1 have tri-methylation of H3K9 on Western blot analysis, When the When SETDB1 lacks H3K9 tri-methylation. This suggests that SETDB1 V600E ;p53 BRAF 1 BRAF , LT., Turner -/- to increase the rate of tumor formation. Human formation. tumor of rate the increase to V600E V600E ;p53;mitfa mutants leads to mosaic rescue of Human melanoma is associated with copy with associated is melanoma Human transgene and transgene 1 , F.F., Ferre 1 , W.L.Lin , mutation are put into a into put are mutation p53 2 eia Oclg, Dana Oncology, Medical 40 minigene and a and minigene mitfa 2 , L.G. Garraway L.G. , 77 2 ,

Talks Talks 1 78 M. Schartl model Genetic background dependent pigment cell tumor formation in a transgenic medaka melanoma Germany; Double transgeniclinesareproducedtostudytheeffectofputative tumormodifiers. focal pigment cell tumors, while tumor onset was unchanged compared to p53 wildtype medaka. Interestingly, on p53 negative background the expression of genes anddifferentiation activation and Mitf- and Bcl-2 levels correlated with a more aggressive stage of the malignancy. several apoptosis of Stat5 that Wefound pathways. relevance transduction signal melanoma-relevant of induction the in-vivo the for and of monitoring first a used was for model The system. Xiphophorus the from known changes molecular oncogenic the xanthoerythrophores, occurred. These pigment cell tumors were shown to perfectly recapitulate invasive background, genetic the on beginning melanoma, Depending uveal melanoma or exophytic and penetrance. less tumors aggressive pigment cell tumors, 100% derived from cell with and pigment stages malignant larval highly at to hyperpigmentation disorders benign develop pigmentary that from ranging wereestablished lines Several promoter. mitf the specific express cell that produced were transgenic medaka stable fish, of transgenic lines a in migration function cell Xmrk tumor analyze further and identified. To sites been ectopichave at survival antiapoptosis, differentiation, cell pigment of suppression proliferation, of stimulation including phenotype neoplastic the responsible establishing are for that events malignant signaling intracellular to critical leads The cells. Xmrk pigment of of transformation Overexpression receptor. factor growth epidermal the of version mutant active constitutively a Xmrk, encodes oncogene melanoma-inducing tumor The development. and transformation cell pigment to lead that mechanisms molecular the study to useful their evaluate easiest to been has model melanoma organism.Xiphophorus whole The required the of context the is in importance clearly are mediators models animal melanoma but models, of culture cell analysis in investigate The to of understood. process partly the only identified, been is have melanomagenesis metastasis and transformation induction, melanoma for Melanoma is a tumor with a very low cure rate once metastasized. Although many genes important Japan Department of Physiological Chemistry I, University of Wuerzburg, Biozentrum, Am Hubland,Wuerzburg,Biozentrum, Am of University I, Chemistry Physiological of Department 41 1 2 , B.Wilde Department of Radiation Genetics, Graduate School of Medicine, KyotoMedicine,University, of School Graduate Genetics, Radiation of Department 1 , J.Laisney 1 , D.Liedtke 1 , S.Meierjohann DA ne cnrl f h pigment the of control under cDNA xmrk xmrk led to the appearance of giant 1 , S.Takeda 2 , Y. Taniguchi SESSION VII Cancer 2 L. I.Partecke 1 3 2 Bagowski P. C. metastatic block and expression a miR-10 behaviour ofpancreaticcancer inhibit antagonists (RAR) receptor acid Retinoic Cancer SESSION VII n acetc acr hc rglts eatss i sprsin f OB ad HOXB3. and HOXB1 of suppression a promisingstartingpointforanti-metastatictherapies. via metastasis is inhibitors) specific (with function or regulates antagonists) RAR (with expression miR-10a of Inhibition which cancer pancreatic in Conclusions: to promotemetastasisformation. HOXB3. and Interestingly, HOXB1 suppression genes, of HOX HOXB1 and of HOXB3 in down pancreatic knock cancer cells specific is by sufficient prevented be can activity anti-metastatic receptor acid retinoic that and This metastasis. block completely and targetexpression miR-10a repress effectively antagonists (RAR) acid retinoid a is miR-10a that formation. demonstrate metastasis further and We invasion inhibit to sufficient is miR-10a of repression that and cells Results: pancreatic in xenotransplantationexperimentszebrafishembryos. human of specimen siRNA resected and and carcinoma. Morpholino Metastatic behaviour lines of primary pancreatic tumours and by cancer cell cell lines was tested carcinoma analysed pancreatic was of miR-10a transfection of role The adenocarcinoma. metastatic in pancreatic miR-10a of levels expression elevated revealed analysis blot Northern Methods: We and function. tumours. pancreatic in formation metastasis of further investigate the upstream and downstream regulatory mechanisms mediator of miR-10a expression important an as (miR-10a) 10a microRNA- identify and embryos zebrafish into fragments tumour human time first the for here lethal of all solid malignancies, largely due to a high frequency of early metastasis. We transplant Background & Aims: Background Greifswald, Germany nvriäslnkm risad Kii fr nee eii A Gefwl, Germany. Greifswald, A, Medizin Innere für Klinik Universitätsklinikum Greifswald Abteilung für Allgemein-, Viszeral-, Greifswald,Thorax- und Gefäßchirurgie, Universitätsklinikum Zoology,Netherlands;Leiden,Integrative Biology,The of of of University Department Institute We show that miR-10a expression promotes metastatic behaviour of pancreatic tumour 3 These findings demonstrate that miR-10a is a key mediator of metastatic behaviour , C-D.Heidecke 1, 1, . . Marques J. I. The infiltrating ductal adenocarcinoma of the pancreas is among the most the among is pancreas the adenocarcinoma of ductal infiltrating The 3 1 , M.Lerch F U Weiss U. F. , 2 2 J M Woltering M. J. , 1 D H Vlecken H. D. , 42 1 , A. Aghdassi A. , 79 2 ,

Talks Talks 80 demonstrates theapplicabilityofzebrafishtomodelcomplexdiseaseprocesses. larvae zebrafish further modulation pharmacological to in phenotypes the of responsiveness The colitis IBD. human and chemically-induced between similarities delineated has study This and metabolism lipid intestinal cellproliferationsuggestinganactivedamagerepairresponseduringinflammation. impaired revealed inflammation increased with associated were phenotypes functional These vascularisation. duringintestinal reduced function intestinal of Studies bythe was prevented administration ofbroad-spectrumantibioticsoranon-steroidalanti-inflammatorydrug. transcription cytokine pro-inflammatory Increased differentiation. cell epithelial intestinal of markers regional to changes and gut larval the within state inflammatory Analysis of gene expression following chemical exposure demonstrated both the induction of an Myd88 forsurvival. Alteration ofmicrobialdiversitywasalsonoted,afeaturetypicalcolitis. wasco- ofinflammation development The Toll-likethe for requirement a was There microbiota. on dependent molecule adaptor receptor inflammation. gut of models murine with shared characteristics key with generated was model colitis zebrafish acute experiments. An murine in To establish doses for inducing colitis in can beinvestigatedtogetherinvivo . zebrafish, larvae were exposed toto standardhave, chemicalscolitis of used date, mainly been studied separately.aetiology Zebrafish providethe a system where these interacting factors to factors environmental and genetic of contributions relative interactions. host-microbe byaberrant necessary and possibly studied sufficient to trigger the escalating commonly ischaracterised inflammation characteristic of IBD. most The dysfunction, (IBD) Epithelial disease bowel Inflammatory Auckland, New Zealand Pathology,of & Sciences,University Medicine Medical The Molecular of of School Department S. Oehlers,M.Flores,K.Okuda,C.Hall,Crosier, P. Crosier Chemical colitismodelsinzebrafish larvae 43 sn ceial-nue clts is colitis, chemically-induced using vivo in Disease Models SESSION VIII We have used antisense morpholino oligonucleotides to disrupt expression of the previously the of reported expression disrupt to oligonucleotides morpholino antisense used have We we havefocussedonskeletalmuscleisoformsoftheTandIsubunits. assembly,sarcomere during Troponincomplex the of role the Tounderstand (hpf). fertilisation accessible and terminal differentiation of muscle easily cells is in larvae the zebrafish trunk is in completed Muscle by 48 zebrafish. hours post the in strategies function of loss using assembly sarcomere in proteins sarcomeric of function the of analysis systematic a undertaken Wehave muscle. overlapping patterns of partially expression, displaying show specificity to cases cardiac, skeletal some slow or in skeletal fast and diversified, have family same the in Genes copies. multiple duplication, genome teleost ancestral the of because zebrafish, In there are few animal models available to study mechanisms underlying the different pathologies. of cardiomyopathy, nemaline myopathy and distal arthrogryposis. Despite the medical relevance forms including muscle, Troponinthe the of diseases genetic human in implicated are complex which anchors the complex to the thin filament by binding to Tropomyosin. Troponin-T,and actin; with heads Allmyosin Troponin-I,of interaction inhibit to ofrequired is which the subunits of three subunits of the Troponin complex are Troponin-C, which is required to bind calcium ions; the Troponin complex, which confers calcium (Actin, dependence to the filaments actin-myosin interactions. thin The to of the Z-disks composed and is M-lines respectively. sarcomere, the In Tropomyosin, contraction, striated muscle, Troponin, of regulation Nebulin, unit of etc) intercalatedcontraction basic with is thick filaments The mediated (mainlyby Myosin),stimuli. attached physiological to response and contraction of regulation for required proteins other several with associated are and fashion regular a highly in organised are filaments contractile myofibrils In Wellcome Trust SangerInstitute, Cambridge, UnitedKingdom M.I. Ferrante, J.E.Collins,D.A.GouldingandD.L.Stemple Disruption ofthe Troponin ComplexLeadstoLossofSarcomeric IntegrityinSkeletalMuscle Disease Models SESSION VIII or I2affectsmaintenanceofsarcomere structure. T3 components fast-twitch the of disruption whereas formation, sarcomere prevents T1 subunit complex sarcomere assembly is impaired. Specifically, we find that disruption of the slow twitch Troponin complex subunit. We find that in thethe absencein ofmutation functionala components of the Troponin characterised and identified also have genome. We zebrafish the of annotation improved with tnnt1, and tnnt3a gene, which is the first animal model of a fast-twitch muscle specific, muscle fast-twitch a of model animal first the is which gene, tnni2 , as well as other genes of the same family that have emergedhave that family same the of genes other as well as tnnt3b, genes are found in found are genes troponin 44 81

Talks Talks fin mutants, we have demonstrated that zebrafish is a relevant model for human distallimb forhuman model relevant isa ofzebrafish number anomalies, andcanserveasausefultoolfortheidentificationofnoveldiseasegenes. ina thatzebrafish demonstrated we have mutated genes mutants, the fin of identification through Thus Hemicentin1. of partner interacting essential an as Fibrillin2 identify and proteins, Fras1 and Frem2 of shedding genetic interaction data demonstrating a role for the proprotein convertase FurinA in membrane suggests a potential involvement in the data, interaction Fraser genetic with along complex. and, vertebrates in Furthermore, protein matrix extracellular this we for role present biochemical and a of description first the is This Hemicentin1. of orthologue zebrafish the encoding gene the in and Fraser complex. Indeed, two of mutants are due to mutations in zebrafish orthologues of the sublamina densa of the basement membrane, as seen in mice mutant for components of the the of blistering display class second epidermis of the fin fold. Electron microscopy demonstrates that fin blisters occur at the level of the of mutants the degeneration, fin embryonic display fin mutants, which can be grouped into two phenotypic classes. Whilst mutants of the first class embryonic zebrafish of number a of cloning positional the describe we Here patients. Fraser in seen syndactyly the underlies limbs the of ridge ectodermal apical the of blistering embryonic other half of Fraser syndrome patients are unknown. Analysis of mouse Fras mutants suggest that matrix protein encoding genes, Approximately 50% of investigated causes are due to mutations in one of two large extracellular chryptophthalmos. and syndactyly by characterized condition congenital a is Syndrome Fraser 82 T. J.Carney disease genes novel potential identifies and syndrome fraser for model zebrafish a reveals analysis Genetic 1 Australia; Genes and Development, IMCB, Biopolis, Singapore; Biopolis, IMCB,Development, and Genes 45 . Critically, we identified that another fin blistering mutant is caused by mutations by caused is mutant blistering fin another that identified we Critically, FREM2. 3 Institute forDevelopmental Biology, University ofCologne, Cologne, Germany 1 , C.Sonntag 2 , N.Feitosa-Martins FRAS1 and FREM2, 3 , M.Hammerschmidt whereas the causative genetic lesions in the 2 Muscle Development, ARMI, Monash, ARMI,Development, Muscle 3 Disease Models SESSION VIII FRAS1 SBDS gene. the in mutation a without patients SDS for strategies therapeutic possible reveal and diagnosis Identification of the SDS-like diseasewhenmutated. product and Sbds interact genetically or if the from distinct therefore is and 15, 3 and have identified candidate genes. In contrast, the zebrafish not been observed in Sbds knockdown embryos. We have have abnormalities However,mapped skeletal hematopoiesis. definitive the dysregulated and development to Similar syndrome inhumans. the Thus, patients. SDS in failure stem cells (HSCs) are normal, while late HSCs are severely reduced, similar to the bone marrow hematopoietic definitive early and primitive insufficiency. Furthermore, pancreas exocrine and immunodeficiency.for mutant zebrafish the describe we Here, have beenidentifiedthatcauseSDS. fully in other genes can result in a clinical SDS been phenotype. However, to date no not other genetic lesions has function its but metabolism elucidated. RNAAs twenty in percent of implicated SDS patients been do not has have mutations product in the protein frequently that syndrome abnormalities. Eighty percent of patients with SDS have mutations in the failure skeletal and insufficiency marrow pancreas exocrine by characterized also bone is SDS leukemia. a in results is (SDS) syndrome Shwachman-Diamond 1 N. S. Trede Ceylon: azebrafish mutantwithShwachman-Diamond syndrome-likebonemarrow failure Disease Models SESSION VIII Department ofOncological Sciencesand Department 1,2 cey, , S. A. Hutchinson opoioijce ebys xii dfcie xcie pancreas exocrine defective exhibit embryos morpholino-injected sbds cey has an SDS-like phenotype characterized by skeletal abnormalities skeletal by characterized phenotype SDS-like an has cey lesion and its potential interaction with 1 , E.Locke cey W ae n h poes f eemnn i the if determining of process the in are We cey. lesion may represent a mutation that leads to an SDS-like an to leads that mutation a represent may lesion ceylon 2 Department ofPediatrics,Department University ofUtah, USA 1 , D.Hu cey lesion represents a new pathway that leads to ) that was originally identified in a screen a in identified originally was that (cey) 1 , B.Demarest sbds could be useful for prenatal sbds gene maps to chromosome 1

46 cey lesion to chromosome SBDS gene. The SBDS SBDS gene, mutations gene cey 83

Talks Talks 84 Santoriello etal.,DisModelMech.20092(1-2):56-67 larvae zebrafish in lost is which AKT,constitutively expressingoncogenicrasandinourzebrafish modelofCostellosyndrome. and ERK activate to ability HRAS restores degradation active oncogene and target it for degradation. number We a less a into HRAS mutant processing for candidates are These enzymes. conjugating ubiquitin of of are expression the investigating of whether increase an blocking show proteosomal syndrome Costello of model zebrafish our in presumably We are (which now investigating the HRAS machinery involved in oncogenic ras ubiquitination. Microarray data of obtained senescence (amechanismthatallowscellstoescapeRASinducedtransformation). PTM novel a between reduces its ability to activate downstream link targets and transform) and oncogene-induced cellular a establish data These in whereas ubiquitinated, proliferating cellsitisnotsignificantlyboundtoubiquitin. is HRAS oncogenic cells senescent in that show results Our in HRAS. oncogenic ubiquitination HRASis with transduced HRAS been checked have which we fibroblasts, brain,oncogenic human parallel proliferating and and In senescence degradation. heart for in targeted and that ubiquitinated shows (PTMs) modifications post-translational cellular senescence, while liver and heart kidney are and hyperproliferating. Analysis brain of shows oncogenic H-RAS model The 2009). al., et Santoriello syndrome, Costello mimics (which specific “resistance” to oncogenic transformation in our model of constitutive HRAS expression type cell this for reasons the examined We expressed. therefore highly is oncogene the if even and ubiquitous a Expression in of oncogenic expressed HRAS in is our models HRAS does not where necessarily lead model to constitutive fashion. cellular a transformation, generated have we Moreover lines. in and levels different at expressed different cell types, using promoters like hsp70, UAS in combination with different GAL4 driver be can HRAS oncogenic where models zebrafish several by the preferential association of Ras mutations with certain types of cancers. WeSome tissues are have more developed affected than others by the expression of oncogenic Ras. This is confirmed IFOM, theFirc InstituteofMolecularOncology, Milan, Italy M. Mione,C.Santoriello,V. Anelli, G.Deflorian,and F. Pezzimenti ubiquitin-mediated proteosomaldegradation ofoncogenicHRAS to linked is syndrome Costello of model zebrafish a in damage DNA and senescence Cellular 47 Disease Models SESSION VIII df. In (dpf). 1 sofa potato; sop potato; (sofa of zebrafish in features characteristic and trimester d-subunit first the of mutation the null (FADS).corresponding ASequences Deformation in Akinesia Fetal death to leading humans, in akinesia fetal a causes subunit AChR an of lack The (NMJ). junction neuromuscular the at (AChRs) receptors The contraction of skeletal muscle is dependent upon synaptic transmission through acetylcholine Abuse and Alcoholism, NationalInstitutesofHealth, Bethesda, USA Alcohol on Physiology,Institute Molecular National of Laboratory Systems, Synaptic Model on J. M.Urban maturation sexual beyond survive to mutant null a enables d-subunit receptor acetylcholine modified A Disease Models SESSION VIII a-actin promoter. AChRs containing these fusion proteins are fluorescent, assemble on the assemble We screened for fluorescent, germ-line transmission of the transgene are current. and synaptic established generate a and line endings, proteins of neuron motor under clusters make fusion membrane, plasma these containing an AChRs of promoter. control the a-actin under loop, intracellular the at fused molecules (d2YFP) protein fluorescent expressing the d2YFP.d2YFP/sop the These expressing of NMJ. synaptic strength. The survival of the transgenic adulthood. animal first to altered delineates had NMJ the and expression transgene survived the drove promoter to requirements foreign a fish, rescued the and In for corresponding transgene gene a animal therapies by mutant rescued been a has of that human case in first lethality trimester the is this knowledge, our of best the To adulthood anddisplaycomplexbehaviorssuchasfeedingbreeding. to Remarkably, grow embryos fish. these type wild to compared when decay slower and time, their wild type siblings. Synaptic currents in these embryos had a smaller amplitude, slower rise The Whitney Laboratory for Marine Bioscience, University of Florida, St. Augustine, USA; sop 2 , K.E.Epley -/- mutants, we expressed modified d-subunits, with one (d1YFP) or twoyellow or one (d1YFP) with d-subunits, modified expressed we mutants, 1 , T. Ikenaga -/- ) leads to the death of embryos around 5 days post-fertilization days 5 around embryos of death the to leads ) 2 , F. Ono -/- embryos can mount escape behavior close to that of that to close behavior escape mount can embryos 2 48 sop -/- fish stably 2 Section 85

Talks Talks 86 Alagille of sufferers amongst deaths of the 50% of haplodeficiency a least with associated at usually disease for a syndrome, account defects heart Congenital ofCellDepartment ofPediatrics, Biology andDepartment Duke University, Durham, USA K. A. NembhardandM.L.Kirby Role ofnotch signalinginearlyzebrafish cardiogenesis Having chosen zebrafish for its genetic tractability, an RNA tractability, an genetic its for zebrafish chosen Having formation. tube myocardial initial after until morphogenesis heart to population second the of contribution the delaying by field cardiogenic the of subpopulations these establishes pathway is coordinated, are fundamental to understanding heart morphogenesis. I propose that the Notch differentiation their how and tube. formed, cellpopulation are populations heart two these when the and how Understanding to asaprogenitor adds and muscle smooth and state myocardium into anundifferentiated differentiates gradually in that maintained is field to heart firstpopulation second the is field heart first The differentiate into myocardium and form the initial, non-proliferating myocardial heart tube. The populations. cell distinct two into segregated is and shown to establish subpopulations within the cardiogenic mesoderm in gene, and these genes encode members of the Notch signaling pathway. This pathway has been window oforgan development. tube. These results show a perfect example of how multifaceted Notch signaling is in a small time whereas that by cell myocardial in increase in the of number extent and timing morpholinos The mechanisms. via separate two embryo by so do zebrafish but tube the myocardial initial the in in number cell myocardial the genes regulate both that demonstrated these down Knocking cardiogenesis. controlling for early candidates as syndrome Alagille with associated members pathway Notch the selecting to led genes pathway Notch 49 . During early mouse and chick embryogenesis, the cardiogenic mesoderm cardiogenic the embryogenesis, chick and mouse early During laevis. Xenopus zfjagged1b zfnotch2 morphants suggests premature differentiation of the second heart field heart second the of differentiation premature suggests morphants suggests an increase in the proliferative index of the initial myocardial zfjagged1b and zfnotch2, in situ in the zebrafish orthologues of orthologues zebrafish the hybridization screen of the of screen hybridization Cardiovascular system Drosophila Drosophila melongaster Jagged1 SESSION IX or the or Notch2 In a forward genetic screen of zebrafish mutants with defects in cardiac valve formation, we mutant formation, lethal valve embryonic cardiac recessive, in the defects isolated with mutants zebrafish of screen genetic forward a In model organism. excellent an become has zebrafish the formation, valve cardiac in involved pathways signaling far little is known about disease causing genes and their respective function. To characterize new group of cardiovascular malformations a high percentage accounts for defects of heart valves, so Congenital heart defects are the leading cause of death amongst newborns. Although within the Internal MedicineIII, Heidelberg, University Hospital Germany I.M. Berger,S.Just,B.Meder, H.A.Katus,W. Rottbauer Bungee – A novel regulatorofcardiacvalve formationinzebrafish Cardiovascular system SESSION IX for thefirsttimethatbungeekinaseplaysanessentialroleduringheartvalveformation. Serine/Threonine kinases are known to be involved in multiple signaling development. pathways. Here we show kinase-domain the layer of the atrio-ventricular canal, suggesting an important role for proper AVC and heart valve inside site the of splice-donor a against phenocopies the directed oligonucleotides antisense morpholino-modified of injection while phenotype, wild-type the restores embryos mutant bng a Thyrosine to an Asparagine within the highly conserved kinase domain. Injection of mRNA in within a gene encoding a zebrafish Serine/Threonine protein kinase, leading to a substitution of the that revealed we cloning positional By endocardial andmyocardialcellswithintheearlyformationof AVC. in formation valve heart of failure and (notch1b endocardial ofimportant has2) expression Altered leaflets. valve heart of precursors (AVC),canal atrio-ventricular the widened cushions, and endocardial arteriosus missing bulbus transgenic of microscopy confocal and analysis circulation.Histological blood of loss progressive a to leading atrium the hours post fertilization (hpf) development including chamber formation proceed normally in and myocardial (versican protein kinase within the embryonic heart is restricted to the endocardial cell endocardial the to restricted is heart embryonic the within kinase protein bungee bng mutant phenotype. By :GFP zebrafish revealed that revealed zebrafish fli:GFP bng mutant hearts dilate and blood regurgitates from the ventricle into and mutant embryos is due to perturbed signalling between signalling perturbed to due is embryos mutant bng bmp4) markers for heart valve development suggests that the in situ hybridisation we demonstrate that expression phenotype is caused by a point mutation point a by caused is phenotype bungee bungee ( mutant hearts have a constricted a have hearts mutant bng bng JH177 bng mutant embryos, around 72 ). While first steps of cardiac of steps first While ). 50 87

Talks Talks is necessarytofacilitateregionalpatterningofthedevelopingaortabylocalextrinsiccues. unexpected molecular diversity within arterial endothelial cells and show that this differentiation aorta, while the single DA in the trunk remains unaffected. Taken together, our studies reveal an endoderm-derived chemokine specifically guides the development of the branched region of the of loss that and LDA , a ligand for ligand a cxcl12b, inactivation of receptor 4a (cxcr4a) specifically labels endothelial cells of thecells anteriorare LDA.genetically distinct. In Accordingly,particular, we targeted find that expression ofendothelial the these that show we Furthermore, mesoderm. plate lateral posterior and anterior the signaling and this In mechanisms elusive. remained the have LDAstudy we show this, that the LDAbranched form in a bidirectional manner from the endothelial cells originating in to of formation contrast the to In contributing molecules midline. the towards migrate mesoderm plate lateral the the from angioblasts in when vasculogenesis, via (DA) develop to aorta shown been has dorsal which single formation, DA a to leading mechanisms and the elucidating in embryo made been the has progress oftwolateral Much of trunk. regions anterior consisting in is“Y”-shaped, (LDA) aortae artery dorsal this embryos, zebrafish In circulation. arterial The aorta traverses the length of the vertebrate body to deliver oxygenated blood to the systemic Worcester, USA 1 88 A. F. Siekmann Chemokine signalingcontributestoregionalpatterningofthefirstembryonic artery USA; Group, Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, a-lnkIsiue o Mlclr imdcn, untr Germany; Muenster, Biomedicine, Molecular for Max-Planck-Institute 51 3 Program in Gene Function and Expression, University of Massachusetts Medical School, Medical Massachusetts of University Expression, and Function Gene in Program 1 cxcr4a leads to a specific defect in anterior LDA formation. We further show that , C.Standley , is expressed in endoderm adjacent to the anterior branches of the of branches anterior the to adjacent endoderm in expressed is cxcr4a, cxcl12b 2 , K.E.Fogarty phenocopies 2 , S. A. Wolfe deficiency. Thus, our results suggest that an that suggest results our deficiency. Thus, cxcr4a 3 andN.D.Lawson Cardiovascular system chemokine (C-X-C motif) 2 imdcl Imaging Biomedical 3 SESSION IX vasculature tomodulateangiogenesisduringintersomiticvessel formation. the where and muscle model the between crosstalk a the regulate to support VEGFa ligand results potent the regulates these miR-1 together Taken meeting. the at presented be will results These mRNA. target the of repression miRNA-mediated block and 3’UTR the in sites target the the regulation of individual target sites in VEGF using target protector MOs (TP). These TPs bind dissecting currently Weare phenotype. down knock miR-1 the suppressed VEGFa of levels the hypothesis we blocked miR-1 function in wild type and VEGFa knock down embryos. Reducing levels in the muscle thus controlling angiogenesis during intersomitic vessel formation. of Toa reporter mRNA. test Takenthis together these results suggested that miR-1 regulates VEGF expression includes three functionally conserved miR-1 target sites in their 3’UTR able of to VEGF regulate pathway effect for expression miR-1 target the sites. the Sequence of Thus analysis components the and searched receptor. we receptor.reporter Third, the studies of upstream revealed functional act that to likely VEGFa is a LOF miR-1 require phenotype overgrowth intersomitic the that revealed function of loss miR-1 and embryos mutant kdrl a using experiments epistasis Second, non-autonomous. is miR-1 of effect the that suggesting phenotype, overgrowth vasculature the type cells labeled with the vascular marker fli1-GFP transplanted into miR-1 MO embryos show wild First, experiments. epistasis and analysis clonal performed we possibilities these between in increase an of reminiscent Toformation. are vessel intersomitic during distinguish signaling cells Notch results of loss a or signaling 3 These VEGF morphants. from miR-1 cells in of cells number 6-9 the to type and wild vessels in intersomitic of area the in increased an caused miR-1 of loss whether MOs non-overlapping two with miR-1 of Inhibition development. vasculature analyzed affected function we hypothesis, this test To development. during vasculature the and muscle the between talk cross the regulate might miR-1 muscle the that suggested hypothesis the observations These genes. of set control a to compared 3’UTRs their in site target 1 miR- muscle the for enriched were vasculature the Conversely,in vasculature. expressed genes development. vivo. Analysis of these in targets revealed muscle that during interactions a miRNAs fraction by regulated of miRNA-target mRNAs them target were of also set large individual expressed a in identified the study of previous Our relevance functional the dissect and targets targetaccelerate and mRNAtranslation decay. However, their identify to challenge a remains it MicroRNAs have recently emerged vivo. as in important modulators modulated gene expression that are repress mRNA signals these how understand to is field the in question fundamental A Cell signaling between different tissues ensures coordinated development during organogenesis. Yale University, New Haven, USA A. J.Giraldez, Y. Mishima,andC.Stahlhut MiR-1 regulatesangiogenesisby modulating VEGF signaling Cardiovascular system SESSION IX 52 89

Talks Talks 1 and S.Schulte-Merker 90 4 B.M. Hogan in zebrafish andhumans The zebrafish with genetic linkage to the human the to linkage genetic with Significantly, we have recently identified a human patient population that develops lymphedema sprouting fromtheposteriorcardinalvein. to an identical failure of lymphangiogenesis and hemangiogenesis at the level of endothelial cell of the lymphatic vasculature, we found that the loss of either lymphangiogenesis. during non-autonomously acts and system, lymphatic the of of pathway precursors earliest migration the lymphangioblasts, the along mesoderm somitic the in expressed is Ccbe1 Ccbe1. protein, to mutants mutant, three one identified found cloning and Positional lymphangiogenesis date. embryonic for required genes identify beboth to can that system alymphatic imaged possess zebrafish that demonstrated recently have We the ligand thatactsthroughVegfr3secreted todirectlymphangiogenesisduringdevelopmentanddisease. a resorption, VEGFC, and Prox-1 factor fat transcription the as homeostasis, such known, are fluid lymphangiogenesis in of roles aspects initial the regulating factors few pivotal very Only metastasis. cancer and response plays immune system vascular lymphatic The USA role invertebratedevelopmentandhumandisease. regulator of lymphangiogenesis in the predicted secreted protein Ccbe1 and highlight its critical zebrafish of function human the in lesions molecular f lncl eeis Aaei Mdcl ete Asedm Te Netherlands; The Amsterdam, Centre, Medical Academic Genetics, Clinical of adooy Toacne, rsu Uiest Mdcl etr Rtedm Te Netherlands; The Rotterdam, Center, Medical University Erasmus Thoraxcenter,Cardiology, Department of Medicine, University of California San Diego, Division of Cardiology,Diego,of San Division Medicine,California of of University Department California, Hubrecht Institute-KNAW & University Medical Centre, Utrecht, The Netherlands; 53 in vivo as well as manipulated genetically. We have performed a forward genetic screen 1 , M. Alders full-of-fluid mutant identifies a secreted protein essential for lymphangiogenesis 1 2 n ypagoeei. hs suis dniy nvl conserved novel a identify studies These lymphangiogenesis. in ccbe1 , F.L. Bos Using a series of new transgenic lines to visualize the development the visualize to lines transgenic new of series a Using 1, 3 , J. Bussmann ee n ta te qiaet uain ihbt the inhibit mutations equivalent the that and gene CCBE1 locus. We demonstrate here that these patients have patients these that here Wedemonstrate CCBE1locus. , to encode a novel predicted secreted predicted novel a encode to full-of-fluid, 1 , M. Witte 1 , N.C. Chi ccbe1 or Vegfc-Ft4 signalling leads 4 , H.J. Duckers Cardiovascular system 3 , R.C. Hennekam SESSION IX 3 Experimental Experimental 2 Department Department 2

model to study vascular myogenesis as well as tumor angiogenesis and other MCs-associated other diseases. and angiogenesis tumor as well as myogenesis vascular study to model cells muscle smooth genetic and cellular useful a vascularas zebrafish establishing vertebrates, higher in found pericytes and the of characteristics functional and molecular morphological, from but other derivatives of the lateral plate mesoderm. Thus, zebrafishprogenitors vascular MCs share many of the endothelial or blood from arise not do vessels trunk the of MCs vascular the ventral that indicate the further data genetic to Our heart. contribute developing the of also tract outflow (VA)they and aorta that anterior and and animals (LDA), developing aortae of dorsal (AMA) lateral arteries the mesenteric around present are MCs vascular that show We zebrafish. in tools genetic and molecular using MCs vascular of origin the investigate we Here, organisms. model available in manipulations experimental and genetic defined performing in the origins of vascular MCs has been hampered by difficulties in observing these cells mural cells (MCs) of around recruitment the the requires system cardiovascular vascular nascent the tree of maturation in and Development a process called vascular myogenesis. Biophysics, UCSF, SanFrancisco, USA 1 M. Santoro Characterization ofvascular mural cellsduringzebrafish development Cardiovascular system SESSION IX Molecular Biotechnology Center, University of Torino,Center,of Biotechnology Italy;University Molecular 1,2 , G.Pesce 1 , V. Mugoni 1 , D. Y. Stainier 2

2 Department of Biochemistry and Biochemistry of Department 54 Understanding in vivo and 91

Talks Talks hs rsls hud ie e isgt o te eeain f lo cls n h vertebrate the in cells blood of generation the on insights new embryo. give should results These of hematopoiesismightoriginatefromhemogenicECs. these if determine to order in from come also subsets EMPs and macrophages primitive mark that lines transgenic to crossed been have fish adult Toswitched them. issue, characterized this and investigate further hours post fertilization, when primitive lineages and EMPs, but not HSCs, have been generated, 27-30 primitive at cells “switched” sorted embryogenesis: we question, Tothat unknown. answer still during is ECs hemogenic arise lineages from arise lineages other these hematopoietic not or Whether EMPs. definitive other and macrophages/erythrocytes three above, stated As hematopoietic genesareturnedon. while down, shut is program genetic endothelial the transition, the during that, show and lines, cells isolate could we during their transition from EC to hematopoietic findings, cells using the flk1:RFP and cmyb:GFPthese reporter Tocorroborate embryo. the in ECs from originate HSCs cells harbor long-term reconstitution when transplanted into irradiated recipients. “switched” that show Moreover,Thus, could (DsRED+). we switched were zebrafish leukocytes adult of 100% (n=30), animals all in and months, 3 at sacrificed were adults transgenic Double recombinase. CRE the by excised is cassette STOP a after DsRED of expression the driving promoter fromhemogenic b-actin born also are ECs in the zebrafish by crossing this CRE line to a switch-reporter line. This latter consists of the of progeny the trace specifically HSCs could we so pan-mesodermic, that not is here used promoter zebrafish, flk1 the in results endothelial cells (ECs). We used similar the Flk1:CRE transgenic line. Unlike in mammals, the zebrafish show we Here ventral sideoftheaortaweregivingrisetoHSCsinmouseembryo. the on cells endothelial vascular that showed nicely have reports technology,recent CRE/LOX independently island blood the using By aorta. the of side ventral the along arise posterior that (HSCs) cells stem hematopoietic from the in arise EMPs zebrafish that shown previously have Wepotential. lymphoid lack that (EMPs) progenitors erythromyeloid committed are precursors generated to produce the adult-type lineages. In both the zebrafish and mouse, the first definitive to embryonic macrophages or erythrocytes. rise Multipotent, or definitive, precursors are subsequently give first precursors primitive, or Monopotent, locations. embryonic different in arising Development of the blood system is complex, with multiple waves of hematopoietic precursors 92 inthe J.Y. cells Bertrand endothelial hemogenic from zebrafish embryo arise hematopoiesis definitive and primitive Both 1 Biology, UCSD, La Jolla, USA; 55 1 , B.Santoso vascular cells. vascular flk1+ Altogether,waves all that indicate data preliminary 1 , N.Chi 2 School ofMedicine, UCSD, La Jolla, USA 2 , D.Traver 1 Hematopoiesis andimmunesystem SESSION X and regeneration/repair of other organ systems that express SDF-1 during embryogenesis and organogenesis embryogenesis involving postnatally.during SDF-1 process express homing that systems the organ of other of analysis regeneration/repair the and in useful prove should line this HSC interact with their which niche by under mechanisms steady the state elucidating and in stress important hematopoiesis be conditions. will In line addition, zebrafish tg(sdf1a:dsRed2) this of creation The systems. mammalian in does it as process transplant homing the during zebrafish the cellular in to contributes axis SDF1-CXCR4 the that indication an was giving transplant after which immediately region kidney fish the to home donor to failed from CXCR4) block (to marrow AMD3100 with Finally,pretreated radiation. myeloablative following expected as RNA expression SDF1a in increase an with correlated which gamma-irradiated were fish when epithelial tubular kidney the in expression positive numbers in increase in showed cells These adulthood. throughout maintained was which cells, dsRed found also we mature, fish the As marrow be roleduring important in organogenesis including the heart, notochord, fins, myofibers and retinal axons. their to understand dsRed2. We have found dsRed positive cells in areas where SDF1 has previously been shown and to zebrafish the transplant, we created an SDF1a transgenic zebrafish by fusing the SDF1a proximal inpromoter to cells niche-establishing HSC to a supportive environment essential for fostering long-term hematopoiesis. To identify potential derived factor-1 (SDF-1), an important mediator that functions to stromal recruit secrete known CXCR4 been expressing have cells HSCs These cells. stromal various contain to thought the is niche How zebrafish. the myeloablative in currently the that subject of much experimentation in the mammalian accomplished systems. In mammals the HSC shown be and zebrafish the in also unknown is engraft can and niche HSC the to way their has find HSC transplant transplanted work (HSC) cell Recent stem cells. hematopoietic hematopoietic of development the zebrafish, The USA in HSC Homing Following Adoptive Cell Transfer in a Zebrafish Model of Bone Marrow of Bone Model T. Zebrafish C.Lund a in Transfer Cell Transplant Adoptive Following Homing Role a Play HSC and Niche in (HSC) Cell Stem Hematopoietic the Establish Cells Expressing SDF-1 Hematopoiesis andimmunesystem SESSION X 1 Transplant, Minneapolis, USA;Transplant,Minneapolis, nvriy f inst, iiin f eiti Hmtlg/nooyBod n Marrow and Hematology/Oncology/Blood Pediatric of Division Minnesota, of University 1 , T. J.Glass i icesnl bcmn a iprat oe i unraveling in model important an becoming increasingly is , rerio danio 1 , J.Tolar 1 2 , L.I.Zon Children’s Hospital Howard Hughes Medical Institute, Boston,Institute, Medical Hughes Howard Hospital Children’s 2 , B.R.Blazar 1 56 93

Talks Talks 94 populate the later that (AGM) Double cells. the aorta-gonad-mesonephros in thymus. and pronephros HSCs definitive marking in HSCs to endothelium haemogenic mouse and zebrafish. from We have generatedtransition a zebrafish Runx1 transgenicthe line, for required also is vertebrates all in HSCs of development the for essential factor transcription a Runx1, endothelium. haemogenic vertebrates. In zebrafish, as in mammals, definitive HSCs are believed to have their origins in the the from HSCs definitive haemogenic endothelium of in a living emergence embryo. The haematopoietic direct program is very conserved the in of demonstration first the provide we Here, endothelium astheoriginofHSCswithinorganism. performed were studies these as can they that show to imaging long-term generate haemogenic endothelial cells that by directly give rise to tracked blood cells in culture. were However, cells mesodermal mouse single of stage endothelial haemogenic transient a of formation through Time-lapse microscopy revealed that bipotential haemangioblasts generate primitive blood cells (the blood. definitive and primitive of generation the in cells endothelial haemogenic of role central precursor endothelial-haematopoietic this debate has recently come from bipotential a to resolution some embryos, mammalian In endothelium. haemogenic the or haemangioblast) progenitors, mesenchymal mesodermal from emerge to cells, proposed been have HSCs controversial. been long has embryo The identity of the cells that give rise to the first haematopoietic stem cells (HSCs) in theThe University developing of Auckland, New Zealand ofMolecular Medicine&Pathology,Department SchoolofMedicalSciences, M. V. Flores,E. Yi NiLam,K.Crosier, C.Hall,andP. Crosier endothelial cellsinzebrafish embryos Live imaging reveals that definitive haematopoietic stem cells emerge directly from haemogenic endothelial cells,providingstronginvivo evidencefora‘haemogenicendothelium’. cells. This demonstrates that haematopoietic cells in the AGM arise from further differentiation of of the cells that initiate expression of EGFP, were red fluorescent mCherry-expressing endothelial imaging of these compound transgenic embryos over 11 hours (24-33 hours post fertilisation), all nuclei of endothelial cells, while HSCs emerging from the AGM express EGFP. During time-lapse 57 rngnc ie express lines transgenic Tg(runx1P2:EGFP/kdrl:nls-mCherry) is expressed in haemangioblasts and in early endothelial early in and haemangioblasts in expressed is (Flk-1) Kdr in vitro, in in vitro live imaging technology that shows evidence for the they still do not definitively identify the haemogenic the identify definitively not do still they Hematopoiesis andimmunesystem . Similarly, the cell fates cell Similarly,the . vitro in Tg(runx1P2:EGFP) n the in mCherry SESSION X critical insightintomammaliantransplantationbiology. already unique genetic and screening advantages mutants of the zebrafish model, such experiments may zebrafish provide with experiments identified in prior genetic screens, transplantation and long-term tumor transplantation competitive assays. By harnessing the including possible, MHC-matched long-term transplantation experiments in zebrafish that have previously not been facilitate will method This haplotypes. zebrafish between genes MHC of test functional first the assay first the represent data These allowing long term HSCs to be 1.41%). distinguished from other hematopoietic progenitors, to and provide 0.83 (range chimerism donor lymphoid mean 1.28% and 8.58%) to 4.58 (range chimerism donor myeloid mean 6.45% only had 6) of 4 (n= 0.92-77.57%) for lymphoid cells. (range Engrafted 10.51% animals receiving and MHC-mismatched cells donor marrow myeloid with for 93.44%) 5.56- fish (range 47.86% 15 of chimerism of donor mean 13 months in engraftment 3 multilineage At had recipients. recipients and MHC-matched donors post-transplant, MHC-mismatched to compared recipients in chimerism donor of percentage and engraftment increases locus 19 chromosome the at haplotypes MHC recipient and donor the matching that demonstrating donors, sibling from cells marrow kidney whole with transplanted were fish recipient MHC-typed genes. specific for amplified products for family single a Utilizing sibling marrow transplants, we identified the four parental MHC haplotypes by transplants. sequencing PCR mismatched and matched performing by haplotypes these of significance functional the testing 19, chromosome on locus MHC core proposed the at haplotypes zebrafish Wedefined frequencies. then HSC activity, mammalian with consistent data suggest that at least 1 out of 65,000 nucleated zebrafish marrow cells contain repopulating by flow cytometric analysis of GFP-expressing donor cells. Statistical analyses of limiting dilution post-transplant, primary and secondary recipients showed multi-lineage engraftment, while minimizing as the mortality measured presumably due to radiation damage of other organs. At 3 identifiedmonths a sublethal radiationWe zebrafish. doseadult ofin 25Gyassay whichreconstitution washematopoietic optimal forlong-term hematopoieticquantitative reconstitution a created we methodologies to assess hematopoietic stem cell (HSC) function are largely undeveloped. Here, reverse screening methodologies leading to the discovery of novel used genetic prominently for mechanisms. the However,study of hematopoiesis, due to the advantages of largegenes and inability scale to immune-match forward donors and and recipients. Despite this, the zebrafish has been (MHC) complex histocompatibility major functional the of understanding poor to due zebrafish Transplantation of hematopoietic cells, tumors or any allogeneic tissue has been hindered in the Y. Zhou J.L.O. de Jong activity inzebrafish Development of an immune-matched transplantation model to detect hematopoietic stem cell Hematopoiesis andimmunesystem SESSION X School, Boston, USA; Cancer Institute, Howard Hughes Medical Institute, Harvard Stem Cell Institute, Harvard Medical 1 Stem Cell Program and Division of Hematology/Oncology Children’s Hospital and Dana Farber 1 , andL.I.Zon 1 , C. E. Burns 2 Clinical Research Program, Boston, Hospital Children's Boston, USA 1 1 , A. Chen 1 , E. Pugach 1 , E. A. Mayhall 1 , A. C.H. Smith 58 1 , H. A. Feldman 95 2 ,

Talks Talks 96 with interference simultaneous However, it. alone or in combination with possible with consistent functions is in attracting which lymphocyte progenitors to epithelium, the thymic thymic primordium. Knock-down the of encapsulating cells the in expressed ligand their only medaka, in receptors chemokine several Of development. thymus of chemokine The epithelial microenvironment. non-functional Given a that lineage. to thymic epithelial owing cells thymopoiesis in cell of failure a with T associated are gene the this in anlage to organ progenitors epithelial these of the transcription commitment factor of cells, colonization progenitor involving lymphoid process, by complex a is Thymopoiesis Freiburg, Germany ofDevelopmentalDepartment Immunology, Max-Planck InstituteofImmunobiology, B. Bajoghli,N. Aghaallaei andT. Boehm Thymus colonizationisunderthecontrolofchemokines andchemokine receptors ccr9 of cooperation the requires colonization thymus that suggesting thymopoiesis, blocked of levels high express to known are that immigrants thymic encompassing populations thymocyte for regulators potential Additionally, we provide evidence that homing isnotabolishedinmicedeficientforeitherCxcl12/Cxcr4orCcl25/Ccr9. u dt sget that suggest data our two the of one microenvironment. results identify genetic networks regulating the homing of lymphocyte progenitors to the thymic . This analysis suggested analysis This Ccr9. 59 and chemokine/chemokine receptor pairs and might explain why thymus why explain might and pairs receptor chemokine/chemokine cxcl12a/cxcr4 are expressed in the thymic rudiment. Moreover, the chemokine Moreover,the rudiment. thymic the in expressed are ccl25a ccl25, we analyzed the expression of chemokines and chemokine receptors during ) is expressed in the developing thymus. Indeed, thymus. developing the in expressed is (bcl11b.2) homologues bcl11b Foxn1 has a key role in the establishing a functional thymic niche. Mutations ccr9a/b , we considered transcription factors that are highly expressed in expressed highly are that factors transcription considered we ccr9, ccr9 r dwsra tre gns of genes target downstream are as a potential candidate. In medaka and zebrafish only zebrafish and medaka In candidate. potential a as bcl11b genes specifically affected thymus homing, but did not abolish ccl25a and ccl25a is a downstream target gene of Foxn1-deficient mice lack the expression Hematopoiesis andimmunesystem specifically and completely and specifically cxcl12a Tkn oehr our together, Taken Bcl11b. ccr9a/b Foxn1. To identify SESSION X receptors and receptors cxcl12a ccl25a/ ccl25a is MHCII secondary (spleen) lymphoid organs. By allowing the study of APCs in their natural environment, reporter lines, allowing an unprecedented view of analyses, APC/T cell FACSinteraction in primary studies, (thymus) and co-expression gene profilingandfunctionalassays.The APC-reporterlineswerecrossedwithavailable Tcell- transgene following the lines the APC-reporter of present we these expressionHere of fertilization. post development, characterization days 5 cell by cells T epithelial thymic during DCs. in observed and MHCII also whichexpression was macrophages of cells, in role B including the subtypes, with APC were identified all Consistent in observed Founders is transgene animals. the of transgenic fluorescent generate to mechanisms thatunderlie APC ontogenyandfunctionalbehavior. zebrafish lines. Because APCs rely upon class II Major Histocompatibility complex(MHCII) MajorHistocompatibility transgenic II new class upon generated genes for antigen presentation, rely we have identified, characterized, and utilized the MHCII we APCs APCs,Because lines. of zebrafish analyses functional and isolation, prospective of adult zebrafish by cytochemical analyses and electron thrombocytic, microscopy. erythroid, To including enable in vivo mammals, imaging, in myeloid and lymphoid cells. found We lineageshave previously identified putative cell DCs in the blood lymphoid organs major all features not possible to other vertebrates. The hematopoietic system of zebrafish is highly conserved and ways in function and development immune of aspects specific examining for potential unique are response immune an have Zebrafish treatments. ofnew of design the phases and etiology disease understanding the for important during compartments interact cell later immune and different produced, the are APCs with when and where how, Defining vivo. dynamic in of studies behavior improving cell in lies cells. biology T APC of naïve understanding of our activators to potent challenge most major the A being DCs with cells, B and (DCs) cells dendritic macrophages, APCs: of types three contains system immune the mammals, In self-antigens. to (APCs), which play key roles in the orchestration of the various forms of immunity and tolerance cells antigen-presenting are immunity adaptive and innate Linking system. immune the of arms adaptive and innate the both of response integrated an requires infection against defense Host Cell andDevelopmental Biology, UCSD, SanDiego, USA V. Wittamer, J.Bertrand,andD.Traver and dendriticcells macrophages lymphocytes, b cells: antigen-presenting study to model transgenic zebrafish A Hematopoiesis andimmunesystem SESSION X DAB transgenic lines should provide new insights into zebrafish immunity and the complex 60 DAB promoter 97

Talks Talks of anon-neurogeniczoneneuralprogenitorcells. Our findings reveal a novel mechanism in which signaling zones. from neurons non-neurogenic underlies the the maintains formation that neurons specific by secreted FGF an identified have areasin inneurogenic-free Moreover,population. progenitor this regulating by hindbrain the in neurogenesis restrict to we progenitors ofneural required is signaling FGFR that shown a population have we approaches, transgenic Using centres. segment identified have studies Our zebrafish hindbrain, in which neurogenesis becomes restricted to zones adjacent to boundaries. spatial and temporal of non-neurogenic zones. theWe have investigated how neuronal differentiation is patterned in the control that formation the in involved signals mechanisms extrinsic the about known is little the neurogenesis, of regulation Despite dissecting zones. in non-neurogenic progress and neurogenic increasing distinct the of formation the by system nervous appropriate location in the nervous system. This is achieved in specific regions of the vertebrate The precise regulation of neurogenesis is critical to generate the correct number of neurons at the 98 R. Gonzalez-Quevedo Negative regulationofneurogenesisby anFGFfromneurons 1 London, UK; iiin f eeomna Nuoilg, ainl nttt fr eia Rsac, il Hill, Mill Research, Medical for Institute National Neurobiology, Developmental of Division 61 2 Department ofCellDepartment Biology, Duke University MedicalCenter, Durham, USA 1 , Y. Lee 2 , K.D.Poss 2 andD.G.Wilkinson Neurobiology I:patterningandbehaviour 1 SESSION XI ie iul akrud dpain VA, poiei rsos (K) otmtr response optomotor (OKR), response optokinetic (VBA), (OMR). adaptation background visual like responses visual basic visual more for “higher” dispensable is in activity its tectum while capture, the prey as of such function role central a for explanation an provides circuitry neuronal This tectum. the by processed selectively be to stimuli smaller allowing inhibition, interlaminar confined toand the superficial dendriticsegregated compartments of tectalare neurons by inputs a local circuitvisual mediating broad Thus, neurons. preiventricular the of bodies cell the to observed instead we case neuronal this activity spreading In from the superficiallocalized. layers ofis the opticstimulus tectum tovisual the deeperthe ones and when silent remain neurons the in experiments in which localization is dependent This on the field. function of receptive GABAergic entire cells located the in the covering neuropil itself broad – is only stimulus visual the when tectum the of layers demonstrated that dendritic processing of visual inputs remains confined to the most superficial Wetectum. the of imaging functional by responses light vivo in recorded and neurons tectal of subpopulations several in or axons retinal the in either GCamP1.6 express to lines driver GAL4 Drosophila antennal lobes to visualize presynaptic and postsynaptic activity. We used a series of indicator, which calcium has proven useful in a variety of preparations including encoded mouse brain slices and genetically a is GCaMP1.6 line. transgenic UAS:GCaMP1.6 a generated we question, this answering toward step first a As dendrites. tectal the of level the at lost is inputs in stratifying neuropil, entire layered of segregation the the if question the raises This layers. non-visual through and visual the of several extend to appear arranged and are larger or layers neurons the tectal to to of perpendicularly majority the of arbors response dendritic main their the Intriguingly, tectum. decrease and the of architecture layered the underlying size principles functional the understanding in interested field Weare response. receptive positive a elicit normally would that the region the within even objects multiple than smaller objects to optimally respond neurons selectivity,wherein size a demonstrated have vertebrates higher of maps. visuotopic parallel 2007). Baier, & Xiao sublayers 2005; three least al., at et into divided (Xiao further is which of of one map tectum, in the terminate a of RGCs layers distinct of re-establishing four subtypes that fashion, discovered previously topographic have We brain. a the in in space visual organized is neurons. projection tectal of retinotectal dendrites the The with connections synaptic of form Axons (RGCs) zebrafish. in cells information ganglion visual retinal for center processing major the is tectum optic The 1 F. DelBene by calciumimaging Segregated processing of visual inputin in the optic tectum by interlaminar inhibition revealed Neurobiology I:patterningandbehaviour SESSION XI Physiology, UCSF, SanFrancisco, USA; calcium responses spread to other layers. On the contrary,the On layers. other to spread responses calcium inhibitory located neuropil these 1 , C.Wyart 2 , A. Muto GABA action was blocked or these neurons were selectively ablated did Moreover, studies in both the tectum of fish and superior colliculus superior and fish of tectum the both in studies Moreover, 1 , EK.Scott 2 MCB, UCBerkeley, USA hs te eioetl erpl otis i o more or six contains neuropil retinotectal the Thus, 1 , EY. Isacoff 2 , H.Baier 1 62 99

Talks Talks 100 Zebrafish neuraltube. the in diversity neuronal the maintain to necessary is zSARA1 if knowing in interested also are We mitosis. across levels signaling correct the ensure to endosomes SARA the using is Notch, characterizing now are We neural state. pathway,signaling or which TGFbeta it’sclarify proliferative analyzing to cargoand endosome zSARA1 the a the the with associate endosomes could during zSARA1 we cells the daughter of cells the inheritance of duringmitosis fate differentiated the tracking segregated by beunequally and and neuroepithelium can the in proliferating ZSARA1 embryo. the Zebrafish zSARA1 the that between of observed development have ratio we morpholinos, the with function modulates protein the down knocking By homologous tothehumanSARA. and Smad binding domain (SBD), and SARA2, without the SBD. We are focusing on SARA1, the vertebrate during FYVE the containing factor SARA1, genes: SARA this two contains on genome zebrafish The study embryogenesis. first a perform to SARA1 zebrafish the studying are We endosomes, unravelinganewmechanismofdirectionalsignaling(Coumailleau,Nature2009). disc (Boekel, Science 2006), but also that in the SOP there is directional Notch trafficking in Sara SARA not only Drosophila ensures that the shown correct have partition lab of our TGFbeta in signaling Studies levels localization. across endosomal mitosis it in confers the that wing domain activated receptors and allow the phosphorylation of the the to close R-Smad the bring to transcription necessary protein adaptor an is activation) receptor factor.for anchor Sara contains transcription a FYVE Smad phosphorylate receptors factors. These can TGFbeta then shuttle to the activated nucleus where they regulate the gene transcription. Sara (Smad signaling, TGFbeta Upon University ofGeneva, Switzerland C. Campos,S. Abke, I.Castanon,M.Fürthauer, M.González-Gaitán SARA1, a TGFbeta signalingadaptor, regulatesproliferation oftheneural precursors 63 Neurobiology I:patterningandbehaviour SESSION XI approaching novelobjects. are they when zebrafish by use eye right preferential the the on take consideration into we report if previous intriguing especially is normally result This may response. IPN fear excessive dorsal suppress the to with function habenula dorsal the of subnuclei lateral left-dominant the connecting tract the suggesting stimulus, light conditioned the of presentation the to response In the fear conditioning, the manipulated fish showed extremely enhanced levels of the freezing conditionally.or constitutively either impaired selectively is habenula dorsal the of subnucleus have we Gal4-VP16, of site target succeeded in establishing the lines in which the neural signal transmission by way the of the lateral of control tetanus under the gene carrying nitroreductase lines the or transgenic gene toxin other with lines such crossing By subnuclei. dorsal habenular the in specifically we Gal4-VP16 expressing hypothesis, line this zebrafish investigate transgenic Tofurther the manners. established sedate have more in it with cope to or be panic may a IPN ventral and in react dorsal to whether danger,as of such the face the in choice that behavioral alternative the in suspecting involved now are we behaviors, involved instinctive adaptive are in neurons more serotonin involvedin its and is raphe the gray the that central and in freezing the as neurons such that the behaviors Consideringdefense and raphe. hindbrain, the the to in project gray IPN central ventral the along axons descending the dorsal IPN send or the ventral to of either subnuclei projects lateral targets. Furthermore, we have specifically recently and discovered that the neurons which in medial the dorsal IPN specifically of the each of habenulae, ratio dorsal size the the in In difference (IPN). (LR) nucleus left-right prominent interpeduncular a the of way by by caused projection habenulo-interpeduncular indirectlyasymmetric prominent a showed we zebrafish, or directly either hindbrain and the within limbic system that pathway connects telencephalic nuclei conduction to the monoaminergic conserved neurons in highly the midbrain evolutionarily an of part are habenulae The K. Kawakami Mishima, Japan, 1 . Agetsuma M. Functional analysisofthehabenulaincontrolfear Neurobiology I:patterningandbehaviour SESSION XI Lab for Devel. Gene Regulation, RIKEN Brain Science Institute, Japan; 2 1 , S.Higashijima H Aizawa H. , 3 OkazakiInstituteforIntegrative Bioscience, Okazaki, Japan 1 T Aoki T. , 3 1 M Takahoko M. , 1 R Nakayama R. , 2 64 National Inst. of Genetics, 1 T Shiraki T. , 1 M Goto M. , 101 1 ,

Talks Talks 102 asymmetry altered how exploring this and nuclei whether might influenceotherneuralpathwaysinthebrain. habenular examining the currently by mediated are is We behavior activity. altered swimming Parapineal-ablated behavior. reduced of exhibited pattern similar also a larvae show to tended fish reversed the adults, As controls. than area swimming smaller a and distance shorter a behavior,covering navigational eye in laterality for in in reduction asymmetry. or significant reversed a with Surprisingly,showed larvae larvae assay larvae reversed wild-type used8-day in observed commonly not to also a were responses differences test, significant and statistically mirror use, activity the motor In for stimuli. tests visual of or variety vibrational werea of asymmetry in those reversed siblings as with their larvae well that from as found indistinguishable behaviors we their studies, prior test In can fish. we adult viable, resulting the are region brain this in asymmetry reversed symmetry with or larvae Because habenula. right the resembling properties with both mediated ablation of the parapineal causes the habenular nuclei to develop more symmetrically, pathway, larvae can be produced that show a left-right (L-R) reversal of these asymmetries. Laser-Nodal-signaling the disrupting By habenula. right the than expression gene different shows and neuropil larger,dense is more nucleus has habenular left adjacent the and anlage pineal the of in larval and adult zebrafish. In >98% of wild-type zebrafish, the parapineal develops to the left behaviors and epithalamus the in asymmetry between relationship the studying been Wehave Embryology, Carnegie, Baltimore, USA M. E.Halpern,L.Facchin Behavioral consequencesofzebrafish epithalamicasymmetry 65 Neurobiology I:patterningandbehaviour SESSION XI regulating BMP receptorendocytosis/trafficking. by development axon SMN in acts Atlastin that suggests which cones, growth in enriched and neurites along all distributed structures endosomal in BMPRI with co-localises partially Atlastin adominant- Finally, primary culture of dorsomorphin. zebrafishreagent, expressing spinalpharmacological neuronsa fromor line prim-5(BMPRI), embryosI enabledreceptor zebrafish BMP us the toof show genetically-modified version that negative a either using pathway BMP the the both Interestingly, pathway. canonical inhibiting by rescued be BMP could morphants Atlastin in observed the defects axon SMN and mobility of inhibitor an as show to functions us Atlastin allowed that have embryos mutant chordino transcripts. and full-length wild-type human in studies and Overexpression zebrafish the both Atlastin with of experiments lack rescue the to performing due by was phenotype this that confirmed have the Wepathway. of BMP/Smad up-regulation significant a by accompanied fragmentation), (microtubule degeneration is preceeded by which abnormal onwards, pathfinding ofpost-fertilisation spinal motorhours neuron72 (SMN) axonsfrom andmobility subsequent axonal larval of decrease drastic a HSP,causes of form early-onset major a in involved protein a Atlastin, of knockdown the that shown spasticity of the lower limbs due to retrograde degeneration of the cortico-spinal tracts. We have HSP is a heterogeneous group of neurodegenerative Toachieve disorders models. mainly during developmentofthezebrafish. characterised by (HSP) paraplegia progressive spastic hereditary in ininvertebrate involved proteins of analyses performed gain-of-function and we lack- degeneration, axons neuron motor neuron spinal of understanding motor the in of progress significant junctions pathfinding ofneuromuscular correct development and proper for (NMJ) required specifically BMPs be the to molecules, signalling shown these were Among stages. embryonic later at Morphogenetic growth Bone synaptic and the as such signalling morphogens, proteins (BMP), Wnt developmental of or Hedgehog, have been identified as guidance families cues and for axonal pathfinding some processes Concomitantly, neurodegenerative pathways. between links various recently unveiled have disorders neurodegenerative several for responsible genes of studies Functional Curie, Paris, France U952/ CNRS 7224, Université P. & M. Curie, Paris, France; 1 J. Hazan Atlastin controlszebrafish spinalmotoraxonarchitecture viainhibitionofthebmppathway Neurobiology I:patterningandbehaviour SESSION XI R Cnr fr eeomna Nuoilg, igs olg Lno, UK; London, College King’s Neurobiology, Developmental for Centre MRC 1,2 , C.Fassier 2 , J.Hutt 1 , S.Scholpp 1 , B.Giros 2 , S.Schneider-Maunoury 3 UMR CNRS 7622, Université P. & M. 66 3 andC.Houart 2 M INSERM UMR 103 1

Talks Talks of preplacodal development to late blastula stage and resolves discrepancies in the literature the origins in discrepancies the regarding theroleofBmpsignaling. resolves traces and stage that blastula model late two-step to development simple preplacodal relatively of a support data These gradient. Bmp the altering without ectoderm of preplacodal into misexpression neurectoderm Conversely,convert can interface. factors competence neural-nonneural a of formation or gradient Bmp the disrupting without specification preplacodal blocks specifically factors competence four all of necessary, as neither signal alone is sufficient to activate preplacodal development. Knockdown ectopic placodal of derivatives production on to the leading ventral competence, side of of zone the the embryo. within anywhere Both development Bmp-antagonism preplacodal and Fgf are within competent cells abutting the neural plate. Misexpression of Fgf8 and Chordin can activate identity preplacodal specify tissue dorsal from secreted Fgf Subsequently,and Bmp-antagonists competence. preplacodal promote to sufficient and necessary are Gata3) and Foxi1 Ap2 (Ap2, ectoderm. Moreover, we find thatsimply establishes preplacodal competence throughout its signaling range within the nonneural four transcription factors co-induced by Bmp priorstudies in zebrafish to support a strikingly gastrulation different model: However, Rather gradient. our than DV a acting along as signaling Bmp a of morphogen, level Bmp discrete identity a of preplacodal readout by specified that is posit models current Most head. the of structures sensory paired the of portions produce that placodes cranial discrete into resolves later domain This plate. neural Preplacodal ectoderm arises during gastrulation as a narrow band of cells surrounding the anterior Cell Biology, University of Iowa, USA 104 B.B. Riley Early requirementsforpreplacodalectodermandsensoryorgandev 1 Biology Department, Texas A&M University, College Station, USA; 67 1 , H-J.Kwon 1 , N.Bhat 1 , R.A.Cornell Neurobiology II:sensoryorgansandregeneration 2 2 Department Department of Anatomy and elopment SESSION XII not beasdifferentitatfirstappeared. may patterning ear inner amniote and zebrafish in Hedgehog of role The vesicle. otic zebrafish Hedgehog that suggest signalling data must be new kept tightly these repressed for Overall the correct tbx1). acquisition (dlx3b,of the dorsolateral markers cell at fates dorsolateral in expanded the of are expense pax2a) (eya1, domains otic ventromedial of markers character: posterior double reported previously the displaying as well as medialised the and ventralised severely is line, ear affected severely most the In ear. the for are lost structures or reduced. Even a otic relatively modest increase dorsolateral in Hedgehog dorsolateral signalling has consequences and embryo, of the throughout expense increased variably is the activity pathway at territories otic ventromedial Hedgehog signalling, Hedgehog of inhibitors in mutations carrying lines of panel a In domains. of expansion an causes activity does not affect dorsoventral and mediolateral otic patterning, an increase in Hedgehog signalling much less obvious. We now show, however, otic that while chick in zebrafish aand loss ofmouse Hedgehog function the are present, if effects, anteroposterior patterning; dorsoventral for required is Hedgehog vesicle, in contrast, In ears. posterior double symmetric mirror to rise gives symmetric double anterior ears, while severe overactivation of the Hedgehog signalling pathway to specify posterior otic identity. A severe or complete loss of Hedgehog signalling Biomedical leads In to zebrafish,mirror Hedgehog signalling from of ventral midline structures is both necessary Department and sufficient and Genetics Biomedical and Science, University ofSheffield, Developmental UK for Centre MRC K. L.Hammond,F. J.M.vanEeden,andT. T. Whitfield in fates the zebrafishcell otic vesicle dorsolateral of acquisition the for required is signalling hedgehog of Repression Neurobiology II:sensoryorgansandregeneration SESSION XII obe uat, h inner the mutants, double ptc2-/- ptc1-/-; 68 105

Talks Talks 106 neuronal progenitorstotheONL,whichremovedthemfrom theproliferativenicheinINL. neuronal progenitors to reach the wild-type number per cluster (Pax6a), and iii) the migration of the of proliferation continued the ii) (Pax6b), cluster each in cell progenitor neuronal first the of proliferation the i) process: regeneration the in points critical three for required are Pax6b and produced light constant of hours proliferating 68 cell after Pax6b, clusters not that but contained Pax6a, an of increased knockdown number Furthermore, wild-type. as of progenitors neuronal neuronal number progenitors. the Thus, half approximately Pax6a contained that clusters cell proliferating yielded Pax6a of knockdown contrast, In cell. progenitor neuronal resulting the to of failed division, cell asymmetric divisions cell initial the the for required However,was after division. Pax6b cell glial Müller until the disrupt glia Müller the in detected not were which Pax6b, or Pax6a either of knockdown contrast, In regeneration. photoreceptor glia-dependent Müller for required is Stat3 that hours, suggesting proliferate, blocked to beginning 51 glia, from glia Müller Müller all and almost the in 31, expressed (16, is which regeneration expression, Stat3 during of Knockdown times respectively). different at expression in increased genes the that identified and experiment microarray gene a performed We which knocksdowntheexpressionofspecifictarget proteins. response, regeneration the during or to prior either retina the into them electroporate then and eye zebrafish adult the into morpholinos inject to technique a developed we amplification, cell progenitor neuronal and proliferation cell The glial Müller of initiation division. the regulating mechanisms cell asymmetric Tocones. molecular outer an and the rods study both the into differentiate they where to (ONL), layer nuclear migrate through that clusters cell cells generate to cellproliferate progenitor to the continue progenitors reentering neuronal neuronal (INL), layer producing nuclear inner and the in cycle population cell stem neuronal adult the represents which glia, Müller the with initiates response regeneration The photoreceptors. only rapidly undergoes apoptosis of both rod and cone photoreceptors and subsequently regenerates adult adult an light-damaged the from use We population. neurons cell retinal stem neuronal of regeneration underlying mechanisms the define to wish We ofBiological Sciences,Department UniversityDame, ofNotre USA D. R.Hyde,T. Bailey, S.C.Kassen,R.Thummel,F. Raycroft, C.M.Nelson light-damaged the of regeneration zebrafish retina during proliferation cell stem neuronal adult Regulating 69 Neurobiology II:sensoryorgansandregeneration zebrafish retina, which retina, zebrafish albino , stat3 , and pax6b, SESSION XII pax6a f agt eonto. e lo on ta te lnr oaiain f ar el i te sensory the in cells hair of polarization planar epithelium influencestheneuron’s the axonarborcomplexity. that found also We recognition. target of kinetics the and topography neuronal of formation the to leading events molecular and cellular has allowed us to formulate an experimental framework to perform an exhaustive analysis of the This process. re-innervation the underlie that ear, events activity-dependent inner key identified have mammalian we the of that to comparable is neuroepithelium whose organ, lateral-line zebrafish the in imaging live Using described. been yet not has process the however,because unclear,remain re-innervation hair-cell permit that mechanisms underlying The damage. after repair can become however,innervated shortly thereafter, granting species, the toorgan full anatomical and vertebrate functional recovery unable Other are birth. mammals damaged neuroepithelia after with because new hair cells cells irreversible throughout their entire sensory lifetimes. is These new hair cochlear death cell hair-cell regenerate sensory to due loss Hearing CDB, CRG, Barcelona, Spain H. Lopez-Schier and A. Faucherre Afferent innervation duringhair-cell regeneration inthezebrafish Neurobiology II:sensoryorgansandregeneration SESSION XII 70 107

Talks Talks 108 lateral lineandthegenesinvolvedindifferentstepsofregenerationprocess. the of cells supporting the in specifically expressed genes the both define to us platform. allow will This Analyzer Genome Illumina the and microarrays using differences, transcriptional at from this transgenic line in the wildtype and in the cells FACssorted Using 1). Marker Cells (Supporting SCM1 called we which line, unableto lateral the of are cells supporting because process divide. regeneration We are the characterizing a in transgenic line deficient expressing GFP exclusively is in the which supporting cells (pho), in events regeneration the the regeneration underlying process in events of the molecular neuromasts. We the unraveling observations have are previously characterized we a lines, mutant morphologic line zebrafish and/ mutant and dyes combining transgenic vital using By larvae, lines. live in transgenic observed be or can days 3 of course the over regeneration Their cells. hair its manner selective highly a in destroying is which copper to exposure and by line dedifferentiation by mainly lateral the in occurs triggered easily be Regeneration can Regeneration cells. supporting line. surrounding the of lateral division the in and ear inner the in cells hair damaged regenerate to ability the have do fish and amphibians, Birds, damage. after them regenerate to unable are they mechanoreceptors, because mammals, the all in cells, deafness for hair cause leading of the is absence or loss the ear, inner the In markers. molecular various of expression the in and ultrastructure in both, neuroepithelia ear’s inner the to similar It is comprised of stereotypically distributed neuroepithelial patches called neuromasts. These are The lateral line in amphibians and fish is a sensory organ that is used to detect water movements. NHGRI, NIH, Bethesda, USA M. Behra andS.Burgess Molecular events duringregeneration inthezebrafish lateral lineaftercoppertreatment 71 Neurobiology II:sensoryorgansandregeneration phoenix mutant background, we are looking SESSION XII phoenix wound healing,reinnervation,scarformation,andotherkeyprocesses invertebratephysiology. epithelial-mesenchymal interactions, melanocyte migration, angiogenesis and lymphangiogenesis, cell types it contains are highly conserved. Thus “barbology” may be a useful system outgrowthfor studyingand regeneration inadult zebrafish. Although the ZMB has no human equivalent, the preliminary observations establish a novel, highly accessible organ system forgenes to studyingbe highly expressed in appendageboth tissues, including reactionchain(RT-PCR) developinganalysesregeneratingoftheseand showedseveral ofZMBs Reverse-transcriptionprocess.this inpolymerase gene key development, barbelsuggestinga of glypican4 for mutant zebrafish Adult injury. to response and investigated members of the Wnt pathway, which are known to be expressed in skin differentiation life-long.To investigate ZMB geneticthe mechanisms barbel The ofoutgrowth regeneration,and havewe injury. is capacity regenerative its to that suggesting old), years 2 response (~ zebrafish senescent in regenerates repeatable also a demonstrating amputation, of rounds multiple after surgery. regeneratehealing, completelyafterstillmonthscould However, of imperfectZMBs 6 these periods to long up after cases, even remodeled some not in were defects and, These rods, tracts. tissue nerve connective misdirected disorganized section, of plane lengths, the had shorter at scars regenerates internal normal, superficially Although imperfect. is regeneration ZMB injury.of structure, missing the of copy well-integrated a produces which regeneration, fin Unlike proximal amputation (n = 350, >95% of barbel length removed), elongating rapidly within 2 weeks catfish) but this capacity has not been tested in zebrafish. Welymphatic showsystem thatin thezebrafish. ZMB Barbel can tissue regenerate is after known to regenerate infurther other investigatedwas vessel speciesbylive this imaging ofof ofthe barbel identity cyprinidscirculation, The (e.g. lymphatic. extendinglarge a recent descriptionsas interpret ofthewe that an vessel alsoclosed-end adjacent but loop,capillary a only not contains ZMB the thatdemonstrate we (fli1:EGFP), cells craniofacial the from sprouts elongates, nerves and ZMB blood supply invade. the Using As transgenic zebrafish with fluorescently labeled endothelial tastebuds. many differentiates epithelium rod. The dorsal epithelium is invaded by melanocytes that are regularly spaced, while the ventral centralU-shapeda formextracellularcondense accumulatestostrandsofthat matrix ZMBthin (30-40days post fertilization) tothe adult (3-6 months). Beginning small,asa epithelial bud, the in immunohistochemistry, types tissuewe have described these scanning-electron and microscopy, the sectioningdevelopment histological and of of the light ZMB maintenancefrom the Usingearly vertebrate. bud stage and adult developmentan the studying for system potential a ZMB the clear,is optically and structure, in simple diameter, in Small nerves. sensory and vessels circulatory cells, pigment buds, taste glands, cells, skin including derivatives, crest neural and mesodermal ectodermal, of number small a contains appendage elongated This ZMB. the called hereafter barbel, maxillary longer a and barbel nasal short a barbels– of sets two develops , rerio Barbels are skin sensory appendages found in fishes, reptiles and amphibians. The zebrafish, 1 E.E. LeClair for adultvertebrate tissuegrowth andrepair system study novel a (ZMB): barbel maxillary zebrafish the of regeneration and Development Neurobiology II:sensoryorgansandregeneration SESSION XII Biological Sciences, DePaul University; 1 , J.Topczewski 2 2 Pediatrics, Northwestern University, Chicago, USA glypican4, show a complete arrest arrest complete a show (knypek) Wnt5a, 72 Wnt5b, and Wnt11. Our Danio Danio 109

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POSTERS

Posters

74 73 76 75 83 82 81 80 79 78 77 Gern cellsandmaternaldeterminants AND ANIMAL USEREGULATIONS INTHEUSA AND EU NURSERYOPERATIONS,FISH, RAISING HUSBANDRYWORKSHOP: UCL AND ZIRC Husbandry Workshop ZEBRAFISH LARVAL REARING AT UCL F. Pezzimenti,Milan,Italy FISH BREEDING AT CAMPUS-IFOM-IEO Z.M. Varga, Eugene,USA THE SOUFFLE GENE IS REQUIRED FOR VESICLE TRAFFICKING DURING OOGENESIS C. Barton,Eugene,USA THE ZIRCNURSERY AND GROW-OUT SECTION J.L. Hakkesteeg,London,UK S. Yang, Beijing,China MUTAGENESIS INZEBRAFISH BY TRANSPOSONGENE TRAP SYSTEMS E.G. Ivaskin,Moscow, Russia THE AT DRUGS NEUROTROPIC OF EARLY PRENERVOUS EFFECTS STAGES OFTELEOSTFISHDEVELOPMENT THE AND SYSTEM SEROTONERGIC M. Kraussling,Wuerzburg, Germany EARLY DURING AND CELL DEVELOPMENT OFORYZIAS LATIPES STEM EMBRYONIC IN STAT3-ACTIVITY OF ANALYSIS K. Goudevenou,Nottingham,UK OF WNT5INTHENON-CANONICAL WNTSIGNALINGPATHWAY DEF6, ZEBRAFISH A NOVEL GUANINE NUCLEOTIDE EXCHANGE FACTOR, EARLY ACTS DOWNSTREAM IN PROTEINS K.M. Hsiao,Chia-Yi, Taiwan MUSCLEBLIND OF DEVELOPMENT STUDIES AND FUNCTIONAL MUSCLE BRAIN, IN INVOLVED GENE H. Pan,Taichung, Taiwan HOMEOBOX NOVEL PHARYNGEAL ARCH DEVELOPMENTINZEBRAFISH A IS IRXL1 Early development andgastrulation F. Müller, Birmingam,UK ZEBRAFISH EMBRYOS VIABLE IN RESULTS FOLLICLES I-II STAGETRANSPLANTATION INTRAOVARIANOF R. Dosch,Génève,Switzerland

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Posters Posters 114 95 94 X.G.Wang, Singapore 93 92 91 90 89 88 87 86 85 84 HW. Han, Taipei, Taiwan NOGO-B THE RECAPITULATES EXPRESSION PATTERN INDIVERSETISSUESINCLUDIND THELIVER AND INTESTINE LINE TG(NOGO-B:GFP) ZEBRAFISH TRANSGENIC (NPM1) ZEBRAFISH G. Deflorian,Milan,Italy DURING NUCLEOPHOSMIN1 SIGNALLING AND WNT OF PRDM5 EMBRYOGENESIS REGULATION THE GENES IN SUPPRESSOR COOPERATE TUMOR THE REGULATION AND FUNCTIONOFSox6INZEBRAFISHSKELETAL MYOGENESIS Signaling T. Kudoh, Exeter, UK OR SMALL EGGS TAIL BUD FORMATION AND SOMITOGENESIS BETWEEN FISH SPECIES WITH LARGE CLOSURE, RING GERM OF DEVELOPMENT HETEROCHRONIC COMPARISONOF A M. Guarienti,Brescia,Italy SUBUNITS (ADAPTOR COMPLEX AP-1) INZEBRAFISH (DANIORERIO) IDENTIFICATION AND FUNCTIONAL CHARACTERIZATION OF A NOVEL Zizioli, Brescia,Italy D. RERIO) (DANIO ZEBRAFISH IN μ1-ADAPTIN AP-1 THE NEURALOF CHARACTERIZATION ZEBRAFISH DURING GENE C. PereiradaCruz,Exeter, EVE1 UK THE DEVELOPMENT OF ANALYSIS FUNCTIONAL CELL I. Roszko,Nashville,USA IN TRILOBITE/VANGL2 COMPONENT PATHWAY WNT/PCP BEHAVIORS UNDERLYING CONVERGENCE AND EXTENSIONMOVEMENTS THE OF ROLE M. Marsal,Barcelona, Spain THE ROLEOFJNKINZEBRAFISHEARLY MORPHOGENESIS X. Tan, Qingdao,P.R.China FLOUNDER(PARALICHTHYS OLIVACEU) FROM FOXD(1,3,5) ANALYSISFUNCTION PATTERNAND EXPRESSION CLONING, B. Wendik, Freiburg, Germany ACTIVATION OFVOXEXPRESSION POU5f1/OCT4 CONTRIBUTES TO DORSOVENTRAL PATTERNING A. Schepis,Stanford,USA BY ALLOSTERIC CONTROL OFα-CATENIN INCELL ADHESION AND MIGRATION TRANSCRIPTIONAL γ- ADAPTIN 109 108 107 106 105 104 103 102 101 100 99 98 97 96 UCIN F H CLAY EE T/PRPL UIG ZEBRAFISH G. Weidinger, DURING Dresden,Germany FTM/RPGRIP1L FEEDBACK MODULATION GENE OF WNT SIGNALING CILIARY BY TROPHOBLASTS.S. Maunoury, Paris,France GLICOPROTEIN-LIKE THE A DEVELOPMENT OF BY REGULATED FUNCTION IS MUSCLE SKELETAL M. Lahne,London,UK IN CONTRACTION-DEPENDENT SIGNALLINGMECHANISM ORGANISATION MYOFIBRIL P. vanDuijn,Utrecht,The Netherlands UNRAVELING PTENFUNCTIONINZEBRAFISHDEVELOPMENT M. Matsuda,Bethesda,USA AND MIGRATION INTHELATERAL LINEPRIMORDIUM NOTCH SIGNALING IS ESSENTIAL FOR COORDINATING CELL FATE, MORPHOGENESIS H. R.Kim,Sheffield,UK ANALYSIS OFTHEHEDGEHOGSIGNALINGPATHWAY INZEBRAFISH F. Rodrigues,Bath,UK AN INVITRO APPROACH FORTHESTUDY OFZEBRAFISHNCMUTANTS A.C. Binot,Liège,Belgium nkx6 GENESCONTROL ENDOCRINEPANCREATIC CELL FATE INZEBRAFISH CATIONICS. Narawane,Bergen, Norway OF ROLE DEVELOPMENTALAMINO ACID TRANSPORTER 1(CAT1) INZEBRAFISH(DANIORERIO) AND DISTRIBUTION SPATIOTEMPORAL M. Milanetto,Padua,Italy IN MOUSE AND ZEBRAFISH GENES EMILIN NOVEL OF FUNCTION AND EXPRESSION OF CHARACTERIZATION T.U. Wagner, Wuerzburg, Germany AND BMP-RECEPTOR 1A CHARACTERISATION OF A FUNCTIONAL PHYSICAL INTERACTION BETWEEN STAT3 S. Jia,Beijing,China AND BALANCINGTGF-BETA AND BMP SIGNALING SIPX IS REQUIRED FOR ZEBRAFISH DORSOVENTRAL PATTERNING BY AUGMENTING P. Porazzi,Milan,Italy NOTCH SIGNALINGREGULATES THYROIDDEVELOPMENTINZEBRAFISH M.J.M. Schaaf,Leiden,TheNetherlands ISOFORM GLUCOCORTICOIDZEBRAFISH THE FUNCTIONALOF THE ROLE RECEPTOR BETA-

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Posters Posters 116 121 120 119 118 117 116 115 114 113 112 111 110 B. Peers,Liège, Belgium IN HEPATO-PANCREATIC DEVELOPMENT IN SOX9B ZEBRAFISH OF ROLE THE DISSECTING V.M. Darras,Leuven,Belgium SEVERELY DISTURBSEMBRYONIC DEVELOPMENTINZEBRAFISH COMBINED KNOCKDOWN OF TYPE 1 AND TYPE 2 AND IODOTHYRONINE DEIODINASES BRAIN IN LIMP2 I. GuerreroGardugno,MexicoCity, PROTEINE Mexico MEMBRANE LYOSOMAL THE NOTOCHORD FORMATION DURINGZEBRAFISHDEVELOPMENT OF ROLE THE F. Imai,Okinawa,Japan UBIQUITIN PROTEASOME SYSTEM IS ESSENTIAL FOR LENS FIBER DIFFERENTIATION T. Mochizuki, Okinawa,Japan AND INTEGRITY EPITHELIAL LENS FIBER DIFFERENTIATION IN DEFECTS SHOWING MUTANT ZEBRAFISH Organogenesis T.T. Luo,Bethesda, USA PATTERN FORMATION EARLYTO CYTOSKELETALDYNAMICS LINKS INKA COMPONENT SIGNALING THE J.M. Topczewska, Chicago,USA MESODERM ROLE OF WNT/²-CATENIN SIGNALING DURING SEGMENTATION LOOP OF THE FEEDBACK PRESOMITIC NEGATIVE J. Topczewski, THE Chicago,USA OF ELEMENT REGULATING WNT/BETA-CATENIN AN SIGNALING IS NOTUM-HOMOLOGUE A.A. Stanton,NewHaven,USA REGULATION OFCHEMOKINESIGNALINGBY THEmicroRNA miR-430 RE. Lindeman,Madison,USA NUCLEAR- IN CENTROSOMAL ATTACHMENT ROLES ESSENTIALAND PRONUCLEAR CONGRESSION WITH LRMP/JAW1 HUMAN TO HOMOLOGOUS POSITIONAL CLONING REVEALS THAT FUTILE CYCLE ENCODES A MATERNAL GENE I. Castanon,Génève,Switzerland DEPENDENT CONVERGENTEXTENSIONDURINGGASTRULATION MEDIATETOWNT- LRP6 WITH INTERACTS 2A/CMG2 RECEPTORTOXIN ANTHRAX GLUTAMATE OF DEVELOPMENT DW. Ali, Edmonton,Canada NORMAL THE FOR SYNAPSES ONZEBRAFISHMAUTHNERNEURONS REQUIRED IS PKCgamma 133 M. Andreazzoli, Pisa,Italy 132 131 130 129 128 127 126 125 124 123 122 D.V. French,Edmonton,Canada Unc119b REGULATES CILIOGENESIS AND LEFT-RIGHT AXIS DETERMINATION THE VENTRAL REGIONOFZEBRAFISHEYE CHARACTERIZATIONVSP,OF INTERACTORAN OF IN EXPRESSED ARMS/KIDINS220 D. Gotti,Brescia,Italy DEVELOPMENT PRELIMINARY STUDY ON THE ROLE OF SIALIDASE NEU3.1 AND NEU4 ZEBRAFISH IN ZEBRAFISH OF PATTERNS CY. Fu,Taipei, EXPRESSION Taiwan AND STRUCTURES TROPONIN IGENES MOLECULAR THE A. Alunni, Gifsur Yvette, France CHARACTERISATION OFNEURAL STEMCELLSINMEDAKA FISHMIDBRAIN OF FORM DOMINANT-NEGATIVEE. Sawatari,Nagoja,Japan OVEREXPRESSING MYOSTATIN DEVELOPS INCREASEDNUMBEROFTHEMUSCLEFIBERS MEDAKA TRANSGENIC GLOMERULAR L. O’Brien,Boston,USA THE TO PODOCYTES OF BASEMENT MEMBRANE ATTACHMENT THE REGULATES PGAF IN SIGNALING IC. Li,Taipei, Taiwan NOTCH BY KLF4 REGULATED NEGATIVELYEMBRYONIC MAMMALIAN INTESTINAL CELL DIFFERENTIATION IS LIKE SUPPRESSOR FUNCTIONING TUMOR 4A FACTOR KRÜPPEL-LIKE ZEBRAFISH M.Watanabe, Osaka,Japan FUNCTIONAL ANALYSIS OF ZREBRAFISH CONNEXIN41.8 FOR PATTERN FORMATION A. Davidson,Boston,USA FROM THEINTERMEDIATE MESODERM RETINOIC ACID ACTS UPSTREAM OF WT1A AND FOXC1A TO SPECIFY PODOCYTES S. Kinoshita,Tokyo, Japan SUPERFICIAL SLOWMUSCLESPECIFICEXPRESSIONS AND CARDIAC INVOLVEDIN MYHM5, GENE HEAVYCHAIN MYOSIN TORARUGU IDENTIFICATION AND FUNCTIONAL ANALYSIS OF THE PROMOTER REGION IN THE R.A. Wingert, Boston,USA THE ZEBRAFISHPRONEPHRICNEPHRON AND ACID THE IRX3B TRANSCRIPTION RETINOICFACTOR TO BY PATTERNEDGENERATE SEQUENTIALLY PROXIMO-DISTAL ARE DOMAINS IN PROGENITORS RENAL 117

Posters Posters 118 145 144 143 142 141 140 139 138 137 136 135 134 HRCEIAIN N FNTOA AAYI O GLUCONEOGENESIS OF P. Gut,SanFrancisco,USA ANALYSIS FUNCTIONAL DURING ZEBRAFISHDEVELOPMENT AND CHARACTERIZATION F. Priller, Berlin,Germany IDENTIFIES MOLECULAR NETWORKSINVOLVED HEARTS INEARLY EMBRYONIC CARDIACMORPHOGENESIS ZEBRAFISH OF PROFILING EXPRESSION GENE K. Hatta, AND Ako-gun, Japan RESPIRATION IN DURING ELEMENTS MOTIONS SKELETAL THEIR FEEDING AND AND TEETH MEDAKA, OF AND STRUCTURES ZEBRAFISH 3D OF IMAGING X-RAY J. Chang,Cambridge,USA REGULATION OFBRAINVENTRICLEINFLATION Y. Quiroz,Liège,Belgium EXPRESSION OFGATA2 AND SPECIFICATION OFTHYROTROPECELLS THE ZEBRAFISH HMG-BOX TRANSCRIPTION FACTOR SOX4B ACTIVATES PITUITARY B.Willems, Singapore OF THEZEBRAFISHCRANIAL SKELETON LRP5 AND ITS PUTATIVE INHIBITOR SCLEROSTIN ARE REQUIRED FOR DEVELOPMENT F. Naye,Liége,Belgium ENDOCRINE PANCREAS MUTATIONSFOR SCREENING EXOCRINE AND OF DEVELOPMENT THE AFFECTING IN CILIOGENESIS FOR ESSENTIAL IS A. Clement,Nashville,USA CDC14B REGULATOR ZEBRAFISH CYCLE CELL THE A.E. Fortunato,Palermo,Italy OF THEHOMEOBOXUNCX4.1 SOMITOGENESISPRIMARYOF DEVELOPMENT AND ROLE MOTORTHE NEURONS: THE E. Maquet,Bruxelles,Belgium IN ZEBRAFISH AS A MODEL TO ROLE STUDY OFTHYROIDDEVELOPMENT ESSENTIAL AN PLAYS CY. Lin,Taipei, Taiwan SIX1A FACTOR CRANIOFACIAL MYOGENESISOFZEBRAFISH TRANSCRIPTION THE M.A. Moriarty, Galway, Ireland IN CARDIOMYOPATHY ZEBRAFISH VENTRICULAR RIGHT ARRHYTHMOGENIC MODELLING 155 154 153 152 151 150 149 148 147 146 157 L.Braeutigam,Stockholm, Sweden 156 THE THALAMUS IN IDENTITY NEURONAL AND GRADIENT NEUROGENETIC REGULATESTHE HER6 J. Schweitzer, Freiburg, Germany OF LONGITUDINAL POSITIONING DOPAMINERGIC DIENCEPHALOSPINAL LATERALAXON COORDINATE HSPGs AND ROBO-SLIT NETRIN-DCC, L.A. Appelbaum, Palo Alto, USA HYPOCRETIN INTERACTS WITH MELATONIN IN REGULATING SLEEP IN BRAIN ZEBRAFISH DEVELOPMENTAL INDUCE R. Creton,Providence,USA SIGNALING CALCIUM DEFECTS AND BEHAVIORAL CHANGES OF MODULATORS Neurobiology I:Patterning E.W. Knapik,Nashville,USA ZEBRAFISH DURING PROTEINS SKELETAL MORPHOGENESIS MATRIX EXTRACELLULAR TRANSPORTS SEC24D I. Porreca,Naples,Italy UNCOVERING NEWGENESINVOLVED INZEBRAFISHTHYROIDDEVELOPMENT F. Langellotto,Naples,Italy SOUL-2, A HEMEBINDINGPROTEIN-CODINGGENEINKIDNEY DEVELOPEMENT FISSURE CHOROID UNDERLYNG G. Gestri,London,UK INTERACTIONS GENETIC FORMATION AND CLOSURE AND CELLULAR T.T. Whitfield, Sheffield, UK VESICLE AND VOLUMEOFTHESWIMBLADDERINZEBRAFISHLARVA MORPHOLOGICALREGULATIONTHE FOR REQUIRED IS ENDOLYMPHNkcc1/Slc12a2 OF OTIC THE IN THE UNDERPIN A.P. Badrock,Parkville, Australia BIOGENESIS INTESTINAL AND CRANIOFACIAL ABNORMALITIES RIBOSOME CHARACTERISTICS OF A IN ZEBRAFISH DEVELOPMENT MUTANT, SETEBOS, WITH EYE, DEFECTS EAIU ATRN BEDN OTOE N ERFS (AI RERIO) (DANIO ZEBRAFISH M.K. Soeffker, Exeter, UK IN OUTCOME COLONIES BREEDING ALTERING BEHAVIOUR REPRODUCTIVE SUBSEQUENT ON IMPACTSEARLY-LIFE ESTROGEN TO EXPOSURE IMPAIRED NEURON DEVELOPMENT TO LEADS 2) GLUTAREDOXIN (ZEBRAFISH GLUTAREDOXINA OF KNOCK-DOWN S. Scholpp,Karlsruhe,Germany

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Posters Posters 120 170 169 168 167 166 165 164 163 162 161 160 159 158 G. Lauter, Huddinge,Sweden PACAP INZEBRAFISHNEURODEVELOPMENT FACTORS TRANSCRIPTION SPECIFIC A. Filippi,Freiburg, Germany OF EXPRESSION IDENTIFIES DISTINCTCATECHOLAMINERGIC GROUPSINZEBRAFISH COMBINATORIAL THE M. SartoridaSilva,Utrecht,TheNetherlands ROLE OFCULLIN-BINDINGDOMAIN ASB11 INNEUROGENESIS Y. Grinblat,Madison,USA A NOVEL ROLEFORZICGENESINTHEDEVELOPINGFOREBRAIN D. Peukert,Karlsruhe,Germany FUNCTION OFTHELHXGENESINTHALAMICDEVELOPMENT OFTHEZEBRAFISH E. Janssens,Leuven,Belgium MATRIX METALLOPROTEINASES DEVELOPMENTAL RETINOTECTAL AXON PATHFINDING IN ZEBRAFISH: A ROLE FOR Y. Yoshimura, Okinawa,Japan MECHANISM REGULATION NEURONAL APOPTOSIS INTHEZEBRAFISHRETINA M. Roussigne,London,UK THE HABENULARNUCLEI NODALLEFT-RIGHTIMPOSES SIGNALLING ASYMMETRYIN NEUROGENESIS UPON C. Yanicostas, Paris,France TARGETING OFOLFACTORY SENSORY NEURON AXON INZEBRAFISHEMBRYOS TERMINALFASCICULATION AND IN ANOSMIN-1A FOR REQUIREMENT ESSENTIAL C.R. French,Edmonton,Canada AND LENSDEVELOPMENT PATTERNINGRETINAL DORSAL-VENTRAL INITIATION OF FOR REQUIRED IS Gdf6a IN 1 DEACETYLASE HISTONE V.T. OF Cunliffe,Sheffield,UK ACTION OF EPIGENETIC CONTROL OFNEURAL PROGENITOR FATE MECHANISM THE DISSECTING L.A. Hale,Eugene,USA MOTONEURON NETRIN SIGNALING IS TELENCEPHALON: REQUIRED FOR DEVELOPMENT OF AN ZEBRAFISH IDENTIFIED ZEBRAFISH ADULT J. Ganz,Dresden,Germany THE IN BY FGF AND BMP SIGNALING NICHES PROLIFERATIONREGULATIONCOMPOSITION AND CELLULAR CHARACTERISTICS, NEUROGENIC THE 183 182 181 180 179 178 177 176 175 174 173 172 171 K. Tanabe, Kobe,Japan PURKINJE OF FORMATIONCELLS INTHEZEBRAFISH CAREBELLUM DENDRITE POLARIZED CONTOLLING MECHANISMS BRAIN’S EARLY ZEBRAFISH X. Zhao,Helsinki,Finland OF DEVELOPMENT THE IN CATECHOLAMINERGIC SYSTEM AMIGO1 OF ROLE THE F. Ono,Bethesda,USA FORMATION OF SPINAL NETWORK DEPENDENT ON DOMAIN-SPECIFIC PAX GENES A. Domenichini,Padua,Italy IN THEDEVELOPMENTOFZEBRAFISHLEFT-RIGHT ASYMMETRIE ANALYSISINVOLVEDMATERNALMOLECULAR AALLELE OF AND EFFECT GENETIC A. Germana’,Messina,Italy SOX-2 EXPRESSIONINTHEBRAINOFDEVELOPINGZEBRAFISH L.D. Knogler, Montreal,Canada BEHAVIOURAL CORRELATES HOMEOSTATIC SYNAPTIC PLASTICITY IN THE DEVELOPING SPINAL CORD AND ITS L. Braeutigam,Stockholm,Sweden ZEBRAFISH NERVOUS SYSTEM LOCALIZATION OF UROTENSIN 1 AND UROCORTIN 3 NEURONS IN THE EMBRYONIC S. Abke, Génève,Switzerland ENDOCYTOSIS AND NEUROGENESISINTHEZEBRAFISHSPINAL CORD JL. Juarez-Morales,Cambridge,UK AND XENOPUS ZEBRAFISH IN FORMATION CIRCUIT NEURAL AND STUDY TO TOOLS GENETICS PMN NEURON, MOTOR PRIMORDIUM C.H. Cheng,Taipei, Taiwan OF REQUIREMENT PROFILE, DEVELOPMENT CHARACTERIZATION OFITSPROMOTERREGION EXPRESSION THE GENE: DX FOR CYCLIN ZEBRAFISH NOVEL A D. Freudenreich,Dresden,Germany ADULT THE IN CELLS PROGENITOR ZEBRAFISH BRAIN NEURAL OF PROFILING EXPRESSION GENE ZEBRAFISH THA IN S. Cote,Montreal,Canada NEUROGENESIS DURING EMBRYO NEUROTRASMISSION OF ROLE PLANAR L. Gordon,Philadelphia,USA THE FOR ROLE CELL POLARITY PATHWAY? ANOTHER FORMATION: SYNAPSE NEUROMUSCULAR 121

Posters Posters 122 196 195 194 193 192 191 190 189 188 187 186 185 184 B. Simões,Faro, Portugal PERIPHERAL NERVOUS SYSTEM DEVELOPMENT PATTERNZEBRAFISH EXPRESSION IN INVESTIGATIONRUNX3 FUNCTION OF AND MY. Law, SaltLakeCity, USA SPATIAL AND TEMPORAL REGULATION OFROBO2SPLICE VARIANTS FOREBRAIN AND INTERNEURON GABAERGIC S IN R. MacDonald,Ottawa,Canada GENES CELL-FATE DLX DEVELOPMENTOF ZEBRAFISH THE OF DIENCEPHALIC ROLE OF COMPONENTS K.M. Kuerner, London,UK GENETIC SPECIFICATION EMBRYONIC THE DISSECTING IN VARIANT MACHRO2A L. Uajardo,Santiago,Chile HISTONE THE DEVELOPMENT OFTHEZEBRAFISH OF FUNCTION N.S. Asuri, Madison,USA BY ROHON-BEARDCENTRAL AXONS EXIT CORD SPINAL ABERRANT ELICIT INTERACTIONS SEMAPHORIN-NEUROPILIN M. Langworthy, Aurora, USA NERVE PERIPHERAL IN 1 CHAIN MYELINATION HEAVY 1 CYTOPLASMIC DYNEIN OF ROLE THE H. Bielen,London,UK PLATE AT EARLY AND PRE-GASTRULA STAGES INZEBRAFISH BMP SIGNALLING IS REQUIRED TO INDUCE CELL FATES IN THE MARGINAL NEURAL GA. Cerda,Cambridge,UK IN INVOLVED FACTORSSPECIFYING CIRCUMFERENTIAL ASCENDING SPINAL INTERNEURONS TRANSCRIPTION OF FUNCTION THE CHARACTERISING J. Blechman,Rehovot,Israel SPECIFICATION AND CIRCUIT FORMATION OF OXYTOCIN - SECRETING NEURONS N. Tiso, Padua,Italy AND FUNCTION INZEBRAFISHCNSDEVELOPMENT INITIATIONAND REGULATION EXPRESSION, GENES OLIG OF ANALYSIS COMPREHENSIVE CEREBELLUM: A ZEBRAFISH ADULT THE J. Kaslin,Dresden,Germany IN MAINTENANCE OF A NOVEL STEMCELL NICHE CELLS STEM C. Danesin,Toulouse, France SPECIFICATION INTHEVENTRAL SPINAL CORD OLIGODENDROCYTE SHH-DEPENDENT SULFATASEANALYSISIN OF FUNCTION 1

208 207 206 205 204 203 202 201 200 199 198 197 P. Washbourne, Eugene,USA PHYSIOLOGICALLY RELEVANT SYNAPSE ATO COMPONENTS OF RECRUITMENT SEQUENTIAL VIVO: IN SYNAPTOGENESIS S. Low, Montreal, Canada TOUCHE`, A NEWTOUCH-UNRESPONSIVE ZEBRAFISH MUTANT A. Nagiel,New York, USA POSTERIOR LATERAL LINEOFTHEZEBRAFISH SPECIFICITY OF AFFERENT SYNAPSES ONTO PLANE-POLARIZED HAIR CELLS IN THE V. Kuscha,Edinburgh, UK AFTER AND BEFORE ZEBRAFISH OF LESION CORD SPINAL THE IN CIRCUIT NEURONAL T. Piotrowski, SaltLakeCity, USA WNT/Β-CATENINREGULATES SIGNALING LATERAL THE OF MORPHOGENESIS LINE M. Kapsimali,Paris,France CELL RECEPTOR BUD TASTE PHARYNGEAL FORMATION FOR NECESSARY IS SIGNALING FGF A. Arrenberg, SanFrancisco,USA OPTICAL CONTROL OFZEBRAFISHBEHAVIOR WITHHALORHODOPSIN B.K. Polok,Sion,Switzerland CELL Bigh3 ISUPREGULATED INREGENERATING ZEBRAFISHFINS HAIR OF SELECTORS STRICT A. Faucherre,Barcelona, Spain AS NEURONS POLARITY AFFERENT LATERAL-LINE: ESTABLISHMENT AND REFINEMENT OF SENSORY INNERVATION IN THE ZEBRAFISH C. Norden,Cambridge,UK ACTOMYOSIN RETINA: FORCES ARE THEPRIMEMOVER ZEBRAFISH THE IN MIGRATION NUCLEAR INTERKINETIC T. Pietri,Eugene,USA SPINAL CORDOFZEBRAFISHEMBRYOS GLUTAMATE DRIVES THE TOUCH RESPONSE THROUGH A ROSTRAL LOOP IN THE OCULAR J.M. Gross, Austin, USA AND PIGMENTATION FOR VERTEBRATE ROLES IN CRITICAL DEVELOPMENT SYNTHESIS IDENTIFY PURINE PAICS NOVO AND DE GART IN MUTATIONS ZEBRAFISH Neurobiology II:Sensoryorgans

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Posters Posters 124 221 220 219 218 217 216 215 214 213 212 211 210 209 B. Blanco,Eugene, USA THE ROLEOFUSH1C DURINGZEBRAFISH OTIC SENSORY PATCH DEVELOPMENT OPSIN MODULATES PHOTORECEPTORS S.L. Renninger, CONE Zurich,Switzerland IN INACTIVATION AVAILABILITY ARRESTIN M. Luz,Dresden,Germany DIFFERENTIATION AND MAINTENANCE OF ZEBRAFISH PHOTORECEPTOR POLARITY B. Kennedy, Dublin,Irleand MUTANT DEFECTIVE PHOTORECEPTORS UNDERLIE INHERITED BLINDNESS IN THE RAIFTEIRI C.M. Maurer, Zurich,Switzerland NOIR MUTANT IMPAIRED ENERGY METHABOLISM LEADS TO REDUCED VISION IN THE ZEBRAFISH K.S. Schaefer, Muenchen,Germany CHARACTERIZATION FUNCTIONAL EVOLUTIONARYAND ZEBRAFISH: IN CLAUDINS MYELIN-SPECIFIC J. Ryan, Montreal,Canada THE WITHIN CHANNELS MOTOR CIRCUITOFZEBRAFISHEMBRYOS CALCIUM VOLTAGE-GATED OF CHARACTERIZATION R. Sanchez-Gonzalez,Salamanca,Spain PROGENITOR ROD OF GENERATION THE OF CELLS MODELS VIVO EX AND VITRO IN A. Germana’,Messina,Italy IN ZEBRAFISH TRKB/BDNF EXPRESSION IN THE HAIR CELLS OF DEVELOPING LATERAL LINE SYSTEM C.J. Huang,Taipei, Taiwan EPHRIN-B1 AND EPHRIN-B3INZEBRAFISHEMBRYOS OF AND INPC-12CELLS SIGNAL REVERSE BY OUTGROWTH NEURITE OF REGULATION DIFFERENTIAL BUD TASTE AND PHARYNGEALG. Gibon,Paris,France EFFECTS MUTATION DEVELOPMENT OL185 ZEBRAFISH THE Y. Omori,Suita,Japan IN CILIOGENESIS AFFECT MUTATIONSPHOTORECEPTOR CELLS REVEALS SCREEN GENETIC GFP-BASED A M. Haug,Zurich,Switzerland METABOTROPIC GLUTAMATE RECEPTORS INTHEZEBRAFISHRETINA

233 232 231 230 229 228 227 226 225 224 223 222 Behaviour A. Rissone,Turin, Italy DEVELOPMENT EMBRYONIC VASCULAR IN NEUROLIGINS IN NEUREXINS OF ROLE SYNERGISTIC C. Campos,Génève,Switzerland REGULATION OFCARDIACDEVELOPMENTBY ZSARA1 IN MORPHOGENESIS VASCULARD. Epting,Mannheim,Germany CONTROLS ELMO1 ZEBRAFISH REGULATOR RAC1 THE C. Tobia, Brescia,Italy ZEBRAFISH IN (EDG1) DIFFERENTIATIONGENE-1 ENDOTHELIAL THE CHARACTERIZATIONOF P. Corti,Pittsburgh, USA VASCULAR REMODELINGISREGULATED BY ALK1 INRESPONSETO BLOODFLOW S. Abdelilah- Seyfried,Berlin,Germany DURING ZEBRAFISHHEART TUBEFORMATION LEFT/RIGHT SIGNALING CONTROLS TISSUE POLARIZATION AND MORPHOGENESIS DERIVED CARDIOMYOCYTES A. Sachinidis,M.Gajewski,Cologne,Germany IN FUNCTION FROM MURINEEMBRYONIC STEMCELLS UNKNOWN WITH TRANSCRIPTS UPREGULATEDSPECIFICALLY TO HOMOLOGUES ZEBRAFISH OF INVESTIGATION BY PATTERNING VASCULAR T. Zygmunt,New York, OF USA MECHANISMS PLEXIND1 SIGNALING CELLULAR AND MOLECULAR Cardiovascular System K. Karlsson,Reykjavik,Iceland THE ONTOGENY OFSLEEP-WAKE CYCLESINZEBRAFISH B. Boyer, Paris,France TASTE PREFERENCESINZEBRAFISHJUVENILES M. Dadda,Padua,Italy AND PERSONALITY IN ADULT ZEBRAFISH USING EARLY CORD LEFT-RIGHT EPITHALAMIC ASYMMETRY SPINAL INFLUENCES BOTH LATERALIZATION THE IN CELLS D. Li,Berkley, USA GLIAN OF INTERSECTIONAL OPTOGENETICS ROLE THE INVESTIGATING 125

Posters Posters 246 245 126 244 241 240 239 238 237 236 235 234 243 242 H GN TA APOC RVAS NOVMN O MK3 I THE IN MEKK3B OF INVOLVEMENT REVEALS A. Urasaki,Mishima, Japan APPROACH TRAP FUNCTIONAL BLOODVESSEL FORMATION GENE THE N. Mercader, Madrid,Spain ZEBRAFISH IN LINE TRANSGENIC EPICARDIAL-SPECIFIC AN OF ANALYSIS AND GENERATION B.M. Hogan,Utrecht,TheNetherlands ARTERIAL SYSTEM Sox18 REGULATES LYMPHATIC DEVELOPMENTINZEBRAFISH P. Lau,Nottingham,UK ANGIOBLAST DEVELOPMENT IN SIGNALLING VEGF FOR REQUIREMENT CELL-AUTONOMOUS A S. Hundt,BadNauheim,Germany OF DEVELOPMENT HEART IN 4 ZEBRAFISH PROTEIN DOMAIN BINDING ATP OF ROLE THE DURING SIGNALING (S1P) C. Detzer, BadNauheim,Germany SPHINGOSINE-1-PHOSPHATE ZEBRAFISH CARDIOVASCULAR DEVELOPMENT OF REGULATION D. Bournele, Athens, Greece ZEBRAFISH MODELSOFCARDIACDEVELOPMENT AND DISEASE B.C. Kirchmaier, Wuerzburg, Germany SYSTEM CONDUCTION CARDIAC IN FAMILYDEVELOPMENT GENE POPDC THE BRAIN OF FUNCTION THE OF CRITICAL FUNCTIONING IS SULF1 FOR S.E. Stringer, Manchester, UK BY AND STRUCTURE PROGRAMMING VASCULATURE SULPHATE ARTERIAL HEPARAN OF FOR TUNING FINE HAEMANGIOBLAST THE F.C. Simões,Oxford,UK OF EXPENSE THE AT FATEPROGRAMME INZEBRAFISH CARDIAC A FAVOURS FGF L4 UPESS EF/L4 INLIG N H DVLPN ZEBRAFISH DEVELOPING THE IN SIGNALLING VEGFC/FLT4 SUPPRESSES DLL4 B. Kennedy, Dublin,Irleand CHEMICAL INHIBITORS OFDEVELOPMENTAL ANGIOGENESIS INTHEEYE P. Vella, Milano,Italy THE ZEBRAFISHEMBRYO DIFFERENTIATIONCELL CD34+ HUMAN TOWARD IN LINEAGE ENDOTHELIAL THE S. Cermenati,Milano,Italy

258 257 256 255 254 253 252 251 250 249 248 247 P. Elks,Sheffield, UK HYPOXIC OF CONTROL MOLECULAR SIGNALLING DURING INFLAMMATION THE INVESTIGATE TO ZEBRAFISH USING A.H. Meijer, DURING Leiden,TheNetherlands SIGNALING RECEPTOR TOLL-LIKESALMONELLA INFECTIONOFZEBRAFISHEMBRYOS AND RESPONSES TRANSCRIPTOME H.B. Taylor, Edinburgh, UK INJURY EMBRYO ZEBRAFISH A MODEL ISP38MAP KINASEDEPENDENT IN RECRUITMENT NEUTROPHIL ENHANCED LPS I. Hess,Freiburg, Germany WATCHING THYMOPOIESIS INSTABLE TRANSGENICZEBRAFISH EMBRYOS F. OF Alcaraz-Perez, ElPalmar, RESISTANCE Spain VIRAL AND IMMUNITY INNATE ZEBRAFISH IN TELOMERASE OF ROLE THE Hematopoiesis andimmunology ZV. Garavito-Aguilar, New York, USA THROUGH FUSION CARDIAC CONTROL OFFIBRONECTINLEVELS FOR ENVIRONMENT PROPER A ENSURES HAND2 H.G. Belting,Basel,Switzerland FORMATION ISV DURING REARRANGEMENTS ENDOTHELIAL FOR REQUIRED IS VE-CADHERIN S. Park,Chicago,USA CARDIAC DEVELOPMENT CHARACTERIZATIONtbx5b, AOF NOVELPARALOG ZEBRAFISH DURING tbx5a, OF A.H. Lim,SaltLakeCity, USA PROMOTES PARACHORDAL FOR REQUIRED VESSEL FORMATIONARE DURINGVASCULAR DEVELOPMENT DCC AND 2 Unc5b RECEPTORS NETRIN THE GREMLIN ANTAGONIST A.K. Hatzopolulos,Nashville,USA PROTEIN DIFFERENTIATION MORPHOGENIC OFEMBRYONIC STEMCELLSTO ATRIAL CARDIOMYOCYTES BONE C. Laufer, Heidelberg, Germany HEART LIGHT CHAIN-1CONTROLSCARDIOMYOCYTECONTRACTILITY INVIVO IN MYOSIN ESSENTIAL CARDIAC OF TERMINUS CARBOXYL THE IN SERINE Hif-1alfa SINGLE A AND Hdac9 A.E. Reyes,Santiago,Chile BETWEEN DEVELOPMENT OFZEBRAFISH INTERACTION FUNCTIONAL 127

Posters Posters 128 271 270 269 268 267 266 265 264 263 262 261 260 259 C.A. Loynes,Sheffield, UK INFLAMMATION RESOLUTION USING TRANSGENIC ZEBRAFISH TO SCREEN FOR SMALL MOLECULE INDUCERS OF L.M. Reaume,Edmonton,Canada PRIMITIVE ERYTHROPOIESIS GATA1REGULATEOF TO UPSTREAM ACT MEIS1 AND COFACTORSPBX HOX THE E. Bresciani,Milan,Italy DIFFERENTIATION ERYTHROCYTES PRIMITIVE IN INVOLVED ARE NUMBLIKE AND NUMB ZEBRAFISH M.A. LýpezMuñoz,Murcia, Spain MOLECULAR AND FUNCTIONAL CHARACTERIZATION OFZEBRAFISHIFNg1-1 M.P. Sepulcre,Murcia, Spain AND FUNCTIONAL CHARACTERIZATION OFZEBRAFISHMD1 AND RP105 MOLECULAR SIGNALING: (TLR) RECEPTORTOLL-LIKE NEGATIVEREGULATIONOF M.P. Sepulcre,Murcia, Spain AND SIGNALING NEW INSIGHTS INTO THE EVOLUTION OF LIPOPOLYCCHARIDE (LPS) RECOGNITION C. Hall, Auckland, NewZealand LIVE IMAGINGDEMAND-DRIVENHAEMATOPOIESIS DURINGINFECTION R. Monteiro,Oxford,UK HAEMATOPOIETIC PROGENITORS REGULATES TIF1gamma ERYTHROIDTHE FROM OUTPUT LINEAGE MYELOID AND D. Aggad, Montpellier, France THE FOR RECEPTORS CONSTITUTE VIRUS INDUCEDIFNSINZEBRAFISH CHAINS CRFB OF COMBINATIONS SPECIFIC N.L. Reynolds,Edinburgh, UK CHARACTERISING ZEBRAFISHDEFENSINS N. Iwanami,Freiburg, Germany IN THYMUSDEVELOPMENT THE ENU-MUTAGENESIS IN ZEBRAFISH IDENTIFIES A SPECIFIC IN REQUIREMENT FOR LSM8 DEVELOPMENT CELL MAST S. Da’as,Halifax,Canada FOR REQUIRED ZEBRAFISH IS SIGNALING NOTCH ADAPTIVE S. Da’as,Halifax,Canada AND INNATE CONSERVED DEMONSTRATE IMMUNE RESPONSES CELLS MAST ZEBRAFISH 283 282 281 280 279 278 277 276 275 274 273 272 P. vanTijn, Utrecht,TheNetherlands ZEBRAFISH MODELSFORFAMILIAL ALZHEIMER’S DISEASE M. Newman, Adelaide, Australia DISEASE PATHOLOGY USING ZEBRAFISH EMBRYOS TO INVESTIGATE GENES IMPLICATED IN ALZHEIMER’S Y. Alvarez, Dublin,Ireland ZEBRAFISH RECAPITULATING EARLY STAGES OF DIABETIC RETINOPATHY IN HYPERGLYCAEMIC Z. Zeng,Edinburgh, UK DEVELOPMENT OFZEBRAFISHMODELSUVINDUCED MELANOMA Disease models IN CELLS BLOOD RED SPL. Hwang,Keelung,Taiwan OF APOPTOSIS ZEBRAFISH EMBRYOS CAUSES TREATMENT Β-LAPACHONE A. Moriyama, Yokohama, Japan HEMATOPOIETIC ONTOGENESIS INMEDAKA INTO INSIGHT AN DIFFERENTIATION; PROGENITORHEMATOPOIETIC DISPLAYS DEFECTIVE WHICH FUJI, BENI MUTANT C-MYB MEDAKA OF CHARACTERIZATION R. Thambyrajah,Nottingham,UK REPORTER GENEEXPRESSIONINHAEMATOPOIETIC AND ENDOTHELIAL CELLS GENE/ENHANCER TRAP BASED SCREEN IDENTIFIES NOVEL TRANSGENIC LINES WITH C. Grabher, Boston,USA CELL FATE THROUGHC-MYB HEMETOPOIETIC COORDINATELY DETERMINE MIR-150 AND MIR-126 ZEBRAFISH C. Grabher, Boston,USA DETECTION INZEBRAFISH A TISSUE-SCALE GRADIENT OF HYDROGEN PEROXIDE MEDIATERS RAPID WOUND S. Louwette,Leuven,Belgium THE ROLEOFRGS18INHEMATOPOIESIS AND MEGAKARYOPOIESIS KA. McMahon,Nottingham,UK TARGET INEMBRYONIC HAEMATOPOIESIS AND ANGIOGENESIS RECEPTORSDOWNSTREAM LIGANDS, AND NOTCH ELUCIDATINGOF ROLES THE T-CELL S.Mitra, Aberdeen, UK OF MARKERS IMPORTANT OF SUBSET INTHEZEBRAFISH(DANIORERIO) ANALYSIS EXPRESSION AND DISCOVERY

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Posters Posters 130 P. Drapeau,Montreal,Canada 296 295 294 293 292 DISC-1IS AN ESSENTIAL MODULATOR OFTHE WNTPATHWAY D.Paquet,Munich,Germany 291 A ZEBRAFISHMODEL OF ALZHEIMER’S DISEASE 290 289 288 287 286 285 284 AIAIN F UAIN RLTD O EEOMNA HMN BRAIN HUMAN DEVELOPMENTAL TO DISEASES USINGZEBRAFISH RELATED MUTATIONS OF VALIDATION M. Poyatos,J.S.Burgos, Granada,Spain ZEBRAFISH EMBRYOS EVALUATIONIN VIVO BIOSAFETY IN OF AND ACTIVITIES OXIDANT VITRO IN ON BASED COMPLEXES RUTHENIUM-NHC OF SERIES A OF PROPERTIES BIOMEDICAL Z. Zhang,Macau,China FISHING FORNEURORPOTECTANTS FROMCHINESEMEDICINE J. Yin, Singapore, China SP117 AND PRP31-AD5,IN A ZEBRAFISHMODEL FORRETINITISPIGMENTOSA FUNCTIONAL CHARACTERIZATION OF TWO HUMAN DISEASE MUTATIONS, PRP31- G. DeRienzo,Cambridge,USA C. Santoriello,Milan,Italy SYNDROME COSTELLO OF MODEL CAUSES CELLULARSENESCENCEIN ZEBRAFISH ADULT PROLIFERATING A CELLS IN H-RASV12 OF EXPRESSION J. Brocher, Singapore A IN DEGENERATIONZEBRAFISH MODEL FORRETINITISPIGMENTOSA PHOTORECEPTOR TO LEADS DEFICIENCY FACTOR SPLICE FUNCTIONAL C. Brusegan,Milan,Italy DISEASES: NEURODEGENERATIVE FOR MODEL ANALYSIS OFGIGYF2, A CANDIDATE GENEFORPARKINSON’S A DISEASE AS ZEBRAFISH F. Geng,Sheffield,UK DEVELOPMENT OFSEMICIRCULARCANALSINTHEZEBRAFISHINNEREAR T. vanBoxtel, Amsterdam, Netherlands LYSYL OXIDASES DITHIOCARBAMATES ARE TERATOGENIC TO ZEBRAFISH THROUGH INHIBITION OF P. Kasher, Amsterdam, TheNetherlands TRNA SPLICING ENDONUCLEAS MUTATION CAUSE PONTOCELEBELLAR HYPOPLASIA D. Sheng,Singapore FOR PARKINSON’S DISEASE MODELA PROVIDES ZEBRAFISH IN LRRK2 OF DOMAIN WD40 THE OF DELETION

306 305 304 303 302 301 300 299 298 297 310 309 308 307

MODELING FRONTOTEMPORAL LOBARDEGENERATION INZEBRAFISH G. Gaudenzi,Milan,Italy ZEBRAFISH, A NEWMODEL TO STUDY RETTSYNDROME H. Dill,Wuerzburg, Germany RETINITIS PIGMENTOSA ANALYSISASSOCIATEDDISEASE FACTORSFOR MODELOF SPLICE AZEBRAFISH IN B.J.C. vandenBosch,Maastricht,TheNetherlands ZEBRAFISH AS A NEWMODEL TO STUDY MITOCHONDRIAL DISEASE Y. vanderVelden, Amsterdam, TheNetherlands THE ROLEOFTUMORSUPPRESSORLKB1INDEVELOPMENT AND CANCER L. Araujo-Bazan, Madrid,Spain VERTEBRATE INVITRO/INVIVOMODEL AAS ZEBRAFISH USING PHOTOPROTECTIVE COMPOUNDS FOR SCREENING NEW ZEBRAFISH DEFICIENT DOK-7 J.S.Müller, Newcastle,UK IN FORMATION EMBRYOS JUNTION NEUROMUSCULAR H-C. Peng,Taipei, Taiwan ACETAMINOPHEN-INDUCED NEPHROTOXICITY INZEBRAFISH L.Wilson, Adelaide, Australia PRESENILIN, INVESTIGATEALZHEIMER’S DISEASERESEARCH TO ZEBRAFISH USING S.A. Sakowski, Ann Arbor, USA LATERALSCLEROSIS AMYOTROPHIC OF MODEL A AS ZEBRAFISH TRANSGENIC G93A-SOD1 DEVELOPMENT IDURONATE-2-SULFATASEOF ROLE AFUNCTIONALNOVEL EARLY ZEBRAFISH IN E. Gibbs, USING Ann Arbor, USA NEUROTOXICITY DYNAMIN-2 FUNCTION IN EMBRYOGENESIS AND SKELETAL MUSCLE FORMATION INDUCED DISEASE F. vanBebber, Munich,Germany ALZHEIMER’S ZEBRAFISH OF ANALYSIS N. Chiarelli,Brescia,Italy OF THEHUMANGENEINVOLVED IN ARTERIAL TORTUOSITYORTHOLOG SYNDROME ZEBRAFISH THE slc2a10, OF EXPRESSION AND CHARACTERIZATION B. Schmid,Munich,Germany E. Moro,Padua, Italy

SCEAE AD P FOR APP AND γ-SECRETASE,

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Posters Posters 132 313 312 311 324 323 322 321 320 319 318 317 316 315 314 ANALYSIS OF COHESIN AND CONDENSIN GENES DURING ZEBRAFISHC. Lillesaar, Neuherberg, Germany DEVELOPMENT polb(ne2385) - A NOVEL ZEBRAFISHMODEL FORNEURODEGENERATION? S. Bretaud,Lyon, France ROLE OFCOLLAGENXVIIIINEYEDEVELOPMENT E. Kabashi,Montreal,Canada FUNCTIONAL CHARACTERIZATION OFMUTANT TDP-43INZEBRAFISH C. Brösamle,Munich,Germany GLIA-NEURONAL INTERACTION AND MYELINATION INZEBRAFISH J. Berger, Clayton, Australia A FISHMODEL FORDUCHENNEMUSCULARDYSTROPHY A. Benini,Brescia,Italy OF MCOLN1,THEGENEMUTATED INMUCOLIPIDOSISTYPEIV CO-ORTHOLOGS PUTATIVE THE OF CHARACTERIZATION AND IDENTIFICATION M. Priyadarshini,Helsinki,Finland UNDERSTANDING PINK1MECHANISMBY GENEEXPRESSION ARRAYS A. Benini,Brescia,Italy FUNCTIONAL CHARACTERIZATION OFTHESLC7A6OSGENEINDANIORERIO PHENOTYPE A. Indrieri,Naples,Italy THE RECAPITULATES MEDAKA IN OBSERVED INMICROPHTHALMIA WITHLINEARSKINLESIONS(MLS)SYNDROME HCCS OF REGULATION DOWN C. Anastasaki, Edinburgh, UK DEVELOPMENT AND ARE SENSITIVETO SMALL MOLECULEINHIBITORS CARDIO-FACIO-CUTANEOUS SYNDROME ALLELES ARE ACTIVE DURING ZEBRAFISH N. Malissovas, Athens, Greece A ZEBRAFISHMODEL OFCHARCOT-MARIE-TOOTH 2D M. Sundvik,Helsinki,Finland CO- PROJECTIONS, TELENCEPHALIC SYSTEM: TRANSMITTERS AND AFFERENT INNERVATION HISTAMINERGIC ZEBRAFISH THE Y-Y. Lin,Cambridge,UK THE EXTENDS PHENOTYPIC SPECTRUMFROMDYSTROGLYCANOPATHY ACTIVITIES TO FAMILYLAMININOPATHY PROTEIN FUKUTIN ZEBRAFISH OF DEPLETION M. Moennich,Dunedin,NewZealand

333 332 331 330 329 328 327 326 325 337 336 335 P.A. Walker, Manchester, UK 334 OE F H PR GNS N AKNO’ DSAE TOOY SN DANIO USING ETIOLOGY DISEASE PARKINSON’SRERIO AS A MODEL IN GENES PARL THE OF ROLE M. Hughes,Manchester, UK ZEBRAFISH AS A MODEL ORGANISMFORLOWE SYNDROME N. Temperley, Edinburgh, UK CELL DEVELOPMENT PIGMENT ALTER THAT MOLECULES SMALL FOR SCREENING CHEMICAL-GENETIC OF INFECTION MODULATE THAT A. Clatworthy, Boston,USA MOLECULES SMALL PSEUDOMONAS AERUGINOSA INZEBRAFISHEMBRYOS OF IDENTIFICATION C. Dooley, Tuebingen, Germany MUTANT, A NOVEL MODEL FORHUMANPIGMENTVARIATION ALBINO ZEBRAFISH THE ANALYSISFUNCTIONAL OF AND CLONING POSITIONAL A. Magnabosco,Padua,Italy THE ZEBRAFISHMODEL FORTHESTUDY OFNEUROTOXINS MECHANISMS T. Hall,Melbourne, Australia IN ZEBRAFISH MODELLING MUSCULAR DYSTROPHY AND EXPLORING THERAPEUTIC STRATEGIES C. Burcklý, Paris,France CMT2A WNT/Β- OF CONTROL FOR FOR CATENIN REQUIRED VERSUSWNT/PCP BALANCE AND CILIOGENESISINZEBRAFISHEMBRYOS PROTEIN CILIARY A IS MODEL NEPHROCYSTIN-4 NEW A G. Bergamin, Padua,Italy KNOCKDOWN: NEUROPATHY? MITOFUSIN-2 ZEBRAFISH J.N. Berman,Halifax, Canada MYELOID HEMATOPOIESIS INTRANSGENIC ZEBRAFISH SURVIVALREPROGRAMS DYSREGULATESCELL AND NUP98-HOXA9 ONCOGENIC G. DeSena,Brescia,Italy CXCR7 AND TUMOR ANGIOGENESIS INZEBRAFISH S. Choorapoikayil,Utrecht,The Netherlands THE ROLEOFPTENINDEVELOPMENT AND CANCER FORWARD CHEMICAL GENETICSCREENING EARLYAN SUITABLEDEVELOPING ZEBRAFISH MELANOMAIN MODELFOR ONSET Cancer S. Noble,Ottawa,Canada

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Posters Posters 134 344 343 342 341 340 339 338 350 349 348 347 346 345 MEDAKA MELANOMAIN SDF1 OF ROLE PROGRESSION CELLPIGMENT AND IN MOVEMENT J. Richardson,Edinburgh, UK MOLECULAR AND GENETIC APPROACHED TO MALANOMA DEVELOPMENT J. Holzschuh,Freiburg, Germany IN CITED1 ZEBRAFISH TRANSACTIVATOR TUMOURIGENIC THE OF ANALYSIS FUNCTIONAL H. Ishizaki,Edinburgh, UK NOVEL IDENTIFY YEAST COMPOUNDS THAT AND SENSITIZEMELANOCYTESFORCELL DEATH ZEBRAFISH IN APPROACHES CHEMICAL-GENETIC V. Anelli, Milan,Italy VALIDATING NOVEL CANCERGENESIN A ZEBRAFISHMELANOMA MODEL PANCREATIC HUMANN A.I. Belo,Leiden,TheNetherlands OF BEHAVIOUR METASTATIC IN CANCER CELLS RIL AND MYSTIQUE LMO7, GENES PDZ/LIM THE FOR AROLE REVEALS MODEL ENGRAFT AZEBRAFISH ZEBRAFISH A IN I.J. Marques, Leiden,TheNetherlands TUMOURS HUMAN PRIMARYXENOTRANSPLANTATION MODEL OF BEHAVIOUR METASTATIC J.K. Frazer, SaltLakeCity, USA HERITABLE ZEBRAFISH TCELL CANCERMODELSFROM A MUTAGENESIS SCREEN XENOTRANSPLANTED FOR E. Rampazzo,Padua,Italy NICHE RELIABLE A HUMAN GLIOBLASTOMA DERIVEDCELLS REPRESENTS BRAIN ZEBRAFISH A.T. Nguyen,Singapore K-RASV12 USINGMIFEPRISTONE-INDUCIBLE CRE/LOXSYSTEM OVEREXPRESSING BY CANCER LIVER ZEBRAFISH ANALYSISGENERATIONOF AND M. Manzoni,Padua,Italy MOLECULAR AND FUNCTIONAL CHARACTERIZATION OF TRIM8 GENES IN ZEBRAFISHK-H. Jeong,Seoul,Korea SEPARATION CHROMATID SISTER IN DURING MITOSIS ROLE CRITICAL A PLAYS PICH ZEBRAFISH AND TELOMERASE OF M. ROLE Anchelin, Murcia, Spain THE STUDY TO MODEL TELOMERE INSTEMCELL BEHAVIOUR NEW A ZEBRAFISH, THE D. Liedtke,Wuerzburg, Germany

363 362 361 360 359 358 357 356 355 354 353 352 351 B. Ayari, Paris,France ADULT ZEBRAFISHTELENCEPHALON INJURED OF REPAIR NEURAL WITH ASSOCIATEDIS EXPRESSION 2 PROKINETICIN V. Parente,Milan,Italy ADULT ZEBRAFISH THE HEART IN RESPONSE INJURY REOXYGENATION AND HYPOXIA E. Sleep,Barcelona, Spain TRANSCRIPTOMICS APPROACH TO INVESTIGATE ZEBRAFISH HEART REGENERATION BY UNAFFECTED T.C. Lund, Minneapolis,USA ARE LENGTH TELOMERE EITHER AGE ORLIMBREGENERATION AND INDANIORERIO TELOMERASE OF EXPRESSION H. Tanaka, Tokyo, Japan ADULT THE ZEBRAFISH OPTICTECTUM IN MARKERS CELL PRECURSOR NEURAL OF PROFILING EXPRESSION IN STRESS HYPOXIC ACUTE M. Marai,Milan,Italy FOLLOWING EXPRESSION ZEBRAFISH GENEADULT HEARTS OF ANALYSIS C. Stewart-Swift,Boston,USA TOOTH, CRANIOFACIAL REPLACEMENT AND SKELETAL FOR MUTANTS INZEBRAFISH SCREEN MUTAGENESIS CHEMICAL ENU GENETIC FORWARD G. Becker, Edinburgh, UK A SMALL MOLECULE SCREEN FOR MOTOR NEURON REGENERATION IN ZEBRAFISH M. Varga, London,UK TECTUM OPTIC ZEBRAFISH THE OF DEVELOPMENT THE IN SIGNALLING WNT CANONICAL C. CiancioloCosentino,Pittsburgh, USA KIDNEY DAMAGE AND REGENERATION INZEBRAFISHLARVAE Regeneration IN FUNCTION AURORA-AH. Jeon,Seoul,Korea STUDYING IN SYSTEM MITOSIS AND DEVELOPMENT POWERFUL A IS ZEBRAFISH L. Zhen,Singapore TRANSGENIC SYSTEMINZEBRAFISH FORMATION OF LIVER TUMOR BY INDUCIBLE EXPRESSION OF C-MYC IN A TET-ON S. Xin,Shenzhen,China EVALUATION OFSMALL-MOLECULEPLK1INHIBITORS USINGZEBRAFISH 135

Posters Posters 136 375 374 373 372 371 370 369 368 367 366 365 364 EEI AAYI O CLNL HRCEITC F AAA NUCLEAR T. Adachi, Nagoya,Japan MADAKA OF ENUCLEATED DIPLOIDIZED EGGS CHARACTERISTIC COLONAL TRSNSPLANTS GENERATED OF FROM THE SOMATIC CELL NUCLEI TRANSFER TO NON- ANALYSIS GENETIC HK. Sehra,Cambridge,UK THE IN HAPLOTYPE CHORI-1073 DOUBLE-HAPLOID ZEBRAFISH MHC AN OF ANALYSIS COMPARATIVE AND ELUCIDATION G. Laird,Cambridge,UK COMMUNITY ZEBRAFISH THE FOR ANNOTATION MANUAL PROVIDING VEGA: HAVANA AND K. Sampath,Singapore DS TRANSPOSONSYSTEM FISHTRAP: INSERTIONALAN MUTAGENESIS THE USING AC/ ZEBRAFISH IN SCREEN K. Naruse,Okazaki,Japan NBRP MEDAKA OF ACTIVITY -AN RESOURCES CDNA FULL-LENGTH MEDAKAESTABLISHMENT OF EARLY IN A.C. Nelson,Cambridge,UK FACTORS T-BOX OF TARGETS IDENTIFY ZEBRAFISH DEVELOPMENT TO APPROACH CHIP-SEQ A Genomics H. Pendeville,Liège,Belgium FNDING miRNAS ABLE TO REGULATE ANGIOGENESIS Y. Takeda, Kobe,Japan THE miRNA-MADIATED GENESILENCING ZEBRAFISH DAZL ENSURES PGC SPECIFIC GENE EXPRESSION THROUGH BLOCKING I. Conte,Naples,Italy AND PAX6 REGULATION VERTEBRATEFOR REQUIRED TARGETINGVIAIS MEIS2 DEVELOPMENT MiR204 EYE A. Ketley, Nottingham,UK MICRO RNA REGULATION OFTHEHEDGEHOGPATHWAY MicroRNA andnon-codingRNAs J-M. Tee, Utrecht,TheNetherlands ASB11 IS A MUSCLESATELLITE CELL MARKERIMPORTANT FORREGENERATION B. Grotek,Dresden,Germany THE ROLEOFNOTCH/DELTA SIGNALINGIN ADULT FINREGENERATION

O.Lee, Exeter, UK 387 O.Konu, Ankara, Turkey 386 385 384 383 382 381 380 379 378 377 376 UAS SYSTEM DEVELOPMENT OF ESTROGEN-RESPONSIVE TRANSGENIC ZEBRAFISH USING GAL4- Emerging technologies THE IN PROFILES EXPRESSION ZEBRAFISH AND HUMAN MODULATED RAPAMYCIN OF META-ANALYSIS Bioinformatics andsystemsbiology K. Howe,Cambridge,UK THE ZEBRAFISHGENOMESEQUENCINGPROJECT: WEBRESOURCES C. Torroja, Hinxton,UK GENOME ASSEMBLY (ZV8) ZEBRAFISH INTO MAPS GENETIC T51 HEAT-SHOCK,AND INTEGRATIONOF MGH R. Kettleborough,Cambridge,UK PROTEIN CODINGGENE THE ZEBRAFISH MUTATION RESOURCE: TOWARDS A TILLING KNOCKOUT IN EVERY S. Sims,Cambridge,UK THE ZEBRAFISH GENOME: AN UPDATE OF MAPPING AND SEQUENCING PROGRESS S. Berger, Clayton, Australia REVERSE GENETICSIN AUSTRALIA: FISHINGFORNOVEL ZEBRAFISHMUTANTS J. Bessa,Seville,Spain IN ZEBRAFISH FACILITATETRANSGENESIS AND THE FUNCTIONAL TOANALYSIS TOOL OF CIS-REGULATORY NEW REEGIONS A VECTOR: (ZED) DETECTION ENHANCER ZEBRAFISH ZEBRAFISH EUROPEAN INTEGRATE R. Geisler, Karlsruhe,Germany TO EFFORT SCALE RESEARCH LARGE A PREDICTIVE - ZF-MODELS AS EMBRYOS S. Scholz,Leipzig,Germany ZEBRAFISH PHOTORECEPTOR ESTERASE INHIBITORS IN TOXICOLOGY RESPONSES ABERRANT MODELS: THE EFFECT OF AZINPHOS-METHYL AND ACETYLCHOLINE TOXICOGENOMIC AND APOTOSIS L. Shine,Dublin,Ireland PROGENITOR MORPHOLOGY CHARACTERISETHEDYINGONEDGE(DYE)MUTANT RETINAL Kondrychyn, Singapore I. ZEBRAFISH IN ENHANCERS THE FOR SCREEN FUNCTIONAL GENOME-WIDE 137

Posters Posters 138 401 400 399 398 397 396 395 394 393 392 391 390 389 388 T.A. Hawkins,London, UK ZEBRAFISHBRAIN.ORG: AN ONLINENEUROANATOMICAL RESOURCE L. Ziv, Ramat Gan,Israel THE GLUCOCORTICOID RECEPTOR MUTATIONPOINT IN WITH ZEBRAFISH ANXIETY-LIKEIN BEHAVIOURINCREASED T. Sýker, Aachen, Germany THE ZEBRAFISHEMBRYO WITH MODEL DISEASE TOTOXICITY SCREENS DO-FROM CAN WHATFISH TINYA R. Brookfield,Nottingham,UK GENE TARGETING INZEBRAFISHBY HOMOLOGOUSRECOMBINATION B. Crawford,Fredericton,Canada MATRIX EXTRACELLULAR DYNAMICS STUDYNG FOR MODEL A AS EMBRYO ZEBRAFISH THE C. Wyart, Berkeley, USA FLUID CONTACTING NEURONSTHAT DRIVELOCOMOTION CEREBROSPINAL SPINAL IDENTIFIES BEHAVIOUR OF DISSECTION OPTOGENETIC S. Baxendale,Oslo,Norway DEVELOPMENTAL MECHANISMSUSINGTHEZEBRAFISH AND PROCESSES DISEASE MODULATORSOF MOLECULE SMALL FOR SCREENING R. Nourizadeh-Lillabadi,Dresden,Germany ASSESS TO EXPOSURE EFFECTSONZEBRAFISHOFNATURAL POP MIXTURES TRANSGENESIS I-SCEI & AC. Oates,Dresden,Germany RECOMBINEERING OSCILLATORY GENEEXPRESSION BAC COMBINING H.B.G. NgSingapore AQUATIC CONTAMINATION BIOMONITORING FOR MEDAKA TRANSGENIC COLOR LIVING OF DEVELOPMENT I. Rothenaigner, Neuherberg, Germany VIRAL CELL TRANSDUCTIONINTHE ADULT ZEBRAFISHBRAIN K. Kobayashi,M.Tanaka, Okazaki,Japan HEAT INDUCTIONOFTHEGENEINMEDAKA EMBRYOS AND ADULT MEDAKA J. Horne,Pleasantville,USA USING IN VIVO ELECTROPORATION FOR TIME-RESOLVED GENETICS IN ZEBRAFISH APOPTOTIC FOR D. Wilkinson, S.S.Gerety, London,UK ROLE NOVEL REVEALS GENES TOXICITY MORPHOLINO ANTISENSE

412 411 410 409 408 407 406 405 404 403 402 A. Sawada,Nashville,USA TO REGULATE CONVERGENCE AND EXTENSIONMOVEMENTS UGLYWNT/PCPWITH PATHWAYINTERACTS PROTEIN DOMAIN SANT DUCKLING, H. Otsuna,SaltLakeCity, USA 4D TIMELAPSE ANALYSIS OFZEBRAFISHEYEDEVELOPMENT PRENYLATIONM. Tada, London,UK THE BY SIGNALLING POLARITY PATHWAY CELL PLANAR OF REGULATION C. Araya, London,UK POLARIZATION AND MORPHOGENESISOFZEBRAFISHNEUROEPITHELIUM MATRIXMESODERM POTENTIALEXTRACELLULAR OF AND ROLE ORGANISING IN C.L. Crespo,Milan,Italy MEDAKA OF MACROPHAGES INVIVO MIGRATION VECTORIAL THE CONTROLS COMPLEX PAR/APKC THE Cell Movement R. Noche,Boston,USA CORD USINGINTOTO IMAGING AND FLIPTRAP SCREEN SPINAL ZEBRAFISH DEVELOPING THE IN LINEAGE NEURON SENSORYPRIMARYA IDENTIFICATIONREGULATORYFUNCTIONALANALYSIS GENE AND OF NETWORK F. Ruf-Zamosjki,Pasadena,Canada ZEBRAFISH EMBRYONS FLIPTRAP IN FORMATION MUSCLE AND SOMITOGENESIS OF IMAGING TOTO IN HIGH-RESOLUTION BYD. Schul,Wuerzburg, Germany PROCESSES CELLULAR MICROSCOPY OF QUANTIFICATION IN-VIVO E. Warp, Berkeley, USA ZEBRAFISH EMBRYO THE OF CIRCUIT SPINAL THE IN CONNECTIVITY DEVELOPING OF IMAGING LIVE GK. Wong, Cambridge,UK NEUROGENESIS INTHEZEBRAFISHRETINA TOWARDS AUNDERSTANDINGMECHANISTIC OF APICALDETACHMENT DURING Live imaging OF M.A. Pardo,Derio,Spain USE THE PROMOTE TO ZEBRAFISH MODEL PLATFORM TECHNOLOGICAL NOVEL A DARENET: 139

Posters Posters 424 R.Lourenço,Lisboa,Portugal 423 422 421 420 419 418 417 416

140 415 414 413 M. Holder, London,UK ABOUT THEHER1/7FEEDBACK LOOP SEGMENTATIONZEBRAFISH THE DECIPHERING UNANSWEREDQUESTION CLOCK: ROLE OFTERRA INVERTEBRATES B. Rajasekaran,Dresden,Germany EMBRYO ANALYSIS OF CELL MOVEMENTS IN THE PRESOMITIC MESODERM OF THE ZEBRAFISH C. Eugster, Dresden,Germany GRADIENT FORMATION INVERTEBRATE DEVELOPMENT AND DISEASE SOMITOGENESISZEBRAFISH TOAAS IN STUDYSYSTEM MODELSIGNALING WNT ZEBRAFISH DURING SIGNALING BK. Liao,Dresden,Germany NOTCH OF SOMITOGENESIS ACTIVATION CONDITIONAL A. Oswald,Dresden,Germany CONSTRUCTING A SYNTHETICSEGMENTATION CLOCKIN YEAST I. DellaNoce,Modena,Italy GENE DURINGZEBRAFISHSOMITOGENESIS dro1/cl2 THE OF CHARACTERIZATION FUNCTIONAL AND ANALYSIS EXPRESSION A. Santos-Ledo,Salamanca,Spain ETHANOL ALTERS THE ESTABLISHMENT OF THE EYE FIELD DURING DEVELOPMENT A. Pistocchi,Milan,Italy IN THEMYOD/MYF5DOUBLEMORPHANTZEBRAFISHEMBRYO INDUCED EARLY EXPRESSION OF MRF4 BUT NOT MYOGENIN RESCUES MYOGENESIS Segmentation L. Beccari,Madrid,Spain EYE FIELSSPECIFICATION OLSOX2-MEDIATED ACTIVATION OF OLSIX3.2 PROMOTES TELECEPHALIC VERSUS R. Marco-Ferreres, Madrid,Spain REGULATORY LOOP BETWEENNEUROD AND SIX6 TIMELYPHOTORECEPTORSDIFFERENTIATIONAON FEEDBACK ROD DEPENDS OF C. Dambly-Chaudière,Montpellier, France CONTROL OFCELL MIGRATION INTHEDEVELOPMENTOFLATERAL LINE

S.Moleri, Milan,Italy 436 435 434 433 432 431 430 429 428 427 426 425 Gene Regulation H hg GN FML I ZBAIH SX EBR WT OVERLAPPING WITH MEMBERS SIX ZEBRAFISH: IN FAMILY EXPRESSION PATTERNS GENE hmgb THE TRANSGENIC BY REVEALED Y. ARE Ono,Tokyo, AS Japan LATIPES REGION ORYZIAS 5’-FLANKING MEDAKACONSTUCTS THEIR OF BY DEVELOPMENT REGULATED CHAIN MUSCLE HEAVY DURING MYOSIN SARCOMERIC GENES OF EXPRESSION SPATIO-TEMPORAL THE H. Schirmer, Porto Alegre, Brasil ZEBRAFISH PGC-1 SIRT1, OF REGULATION N. Wittkopp, Tuebingen, Germany EMBRYONIC DEVELOPMENT AND SURVIVAL ZEBRAFISH FOR ESSENTIAL ARE EFFECTORSDECAY mRNANONSENSE-MADIATED M. Gering,Nottingham,UK TRANSCRIPTIONAL REGULATION OFTHESTEMCELL/PROGENITOR GENEc-myb A. Anusha Amali, Singapore LACKING GROUP 4PARALOG OFHOXGENES ABNORMAL NEUROGENESIS, ANGIOGENESIS AND HAEMATOPOIESIS IN ZEBRAFISH A. Archer, Stockholm,Sweden IN DEVELOPMENT ZEBRAFISH DURING DOWN KNOCK (LXR) LIVER-X-RECEPTOR OF EFFECT M. Andersson-Lendahl, Stockholm,Sweden ABSENCE AND PRESENCEOFLIGAND 17β-ESTRADIOL RECEPTORS EXPRESSED IN EMBRYONIC AND ADULT ZEBRAFISH IN E.H. Barriga,Santiago,Chile ROLE OFHif-1alphaINTHENEURAL CRESTCELLSMIGRATION M.R. Bogo, Alegre, Brazil TO MODULATE THESIRTUINS EXPRESSION IN POTENTIALLYDRUGS NEW TOSCREEN AORGANISM AS ABLE MODEL ZEBRAFISH ascl1a AND ngn1 GENES PRONEURAL R. Madelaine,Toulouse, France THE OF ZEBRAFISH ACTIVITY THE STUDING PR. Jusuf,Cambridge,UK THE AT RETINA ZEBRAFISH EXPENSE OF ALL EXCITATORY THE INCELL FATES FATES CELL INHIBITORY DETERMINES PTF1A YEVRTO I LVR OF LIVER IN BYRESVERATROL PPARγ AND α 141

Posters Posters 142 449 448 447 446 445 444 443 442 441 440 439 438 437 N.M. Feitosa,Cologne,Germany THE ROLEOFHEIMICENTIN INFINDEVELOPMENT THE WITH FISH J-P. Levraud,JouyenJosas,France TELEOST OF FAMILY GENE HALLMARKS OF ANTIVIRAL RESTRICTIONFACTORS DIVERSIFIED HIGHLY A FINTRIMS: A. Gautier, Rennes,France EXPRESSION SPECIFICALLY INFISH TRANSGENE TESTIS DRIVES GENE GSDF ZEBRAFISH THE OF PROMOTER PROXIMAL THE O. Armant, Eggenstein,Germany EMBRYOS MEDAKA GENOME WIDE EXPRESSION IN ANALYSIS OF TRANSCRIPTION FACTORS REPEAT IN ZEBRAFISH TANDEM OF H. Mitani,Tokyo, Japan NUMBER BETWEEN MITOCHONDRIAL DNA AND COLD ADAPTATION RELATIONSHIP THE S.Y. Ler, Singapore CHARACTERIZATION OFTHEHOXD4NEURAL ENHANCER P. Coutinho,Edinburgh, UK DECIPHERING THEPAX6 TRANSCRIPTIONNETWORK E. Kague,Philadelphia,USA THROUGH ZEBRAFISHTRANSGENESIS FUNCTIONALLY CONSERVED CIS-REGULATORY ELEMENTS OF COL18A1 IDENTIFIED E.E. Pashos,Philadelphia,USA MUTATIONAL ANALYSIS OF PTF1A FUNCTION AND TRANSCRIPTIONAL REGULATION L. Kwok,Denver, USA PRDM GENESINZEBRAFISHCRANIOFACIAL DEVELOPMENT A. Chakraborty, Miyazaki,Japan UPREGULATION OFP53-MODULATORY NUCLEOLARPROTEINS RIBOSOMAL PROTEIN L11 (RPL11) DEFICIENCY IN ZEBRAFISH LEADS TO A SELECTIVE J. Ikle,Denver, USA SIGNALING MECHANISMSREGULATING LOWERJAW DEVELOPMENT IN EXPRESSION GENE T. CarneiroBrandãoPereira,Porto Alegre, Brasil SIRT1 INCREASES ZEBRAFISH LIVER TREATMENT ALCOHOL LONG-TERM Z. M. Varga in theUSAandEU Zirc and UCL Husbandry Workshop: raising fish, nursery operations, and animal use regulations Husbandry Workshop 1 euaoy vrih. hr wl b to re peettos y h ognzr t poie an provide to organizers with timeforsharingconcernsanddiscussion. the by presentations brief two overview of regulations be in the USA will and in the There EU. oversight. use regulatory animal in developments recent address to devoted be will workshop the of part second A a and generaltroubleshooting. hosting are (UCL) London three brief presentations and ample time for College discussion, exchange – of two information, tips be and will tricks, nursery. There fish University successful a for needs crucial most and the highlight to workshop (ZIRC) Center Resource International To address this concern and help improve nursery performance and survival rates, the Zebrafish juveniles asthemainareaofconcernintheirdailyfishmaintenanceoperation. caretakers, fish facility attending managers, the and of principal investigators majority identified a the Madison, nursery in and Conference raising Genetics larvae/ and Development Zebrafish At the 2008 Zebrafish Husbandry & Facility Management Workshop held during the International College London, UK ZIRC, University of Oregon, Eugene, USA; Eugene, Oregon, of University ZIRC, 1 andC.Wilson 2 2 Anatomy and Developmental Biology,University Developmental and Anatomy This part of the workshop will also be followed 143

Posters Posters 144 hasbeen larvae ofzebrafish themaintenance and breeding the facility standardized tothelevelthatwehave80%survivaladulthoodandhomogeneoussize. fish our in Now release themindifferentnumbersandfeedlarvaeaccordingtotheirsizenotforage. to tried have Moreover,I survival. 70-80% a gives that cleaning and feeding for schedule a up not to be necessary. I have devised a way to clean several the nursery tanks and from the excess larvae food and set for food dry of providers of types different tried I breeding, of of maintenance method and our Todevelop breeding in experience my explain to zebrafish larvaeusedattheCampus-IFOM-IEOofMilan. like would I presentation this In COGENTECH (Consortium forGenomic Technologies), Milan, Italy F. Pezzimenti Fish breedingatCampusIFOM-IEO 73 Artemia Artemia salina. Initially the larval diet included also paramecia, but I then found out Husbandry Workshop wn t te obnto de ad rtcl floe, e urnl hv sria rts of rates survival have currently we followed, protocols 85%. and diet combination the to the that Owing found was it compared, were - both of combination diet produced higher survival rates combination as well as a much faster increase a in body mass. and live, dry, - diets the fry popular of raising for types for different protocols three where following trial a and From culture cysts. shrimp paramecium brine of good de-capsulation a keeping as rates well flow water as increasing accordingly and size their to according appropriately feeding as such implied to standardized protocols. In order to achieve the best survival rates various methods have been This presentation is aimed to show how zebrafish larvae are successfully raised at UCL according UCL Fish Facility, Division ofBiosciences, University ofLondon, UK J. L.Hakkesteeg Zebrafish Larval RearingatUCL Husbandry Workshop 74 145

Posters Posters 146 successful nurseryingeneral: foodsize,quantity, andwaterquality. discuss will We lines). ZIRC-specific procedures; however,mutant we will also emphasize the three components(some that ensure a 70% and types) (wild 80% between ranges rate survival larvaeare our zebrafish grow, fish the Because average as our changes flow water and diets of progression zebrafish. incremental an given rearing for workflow general the streamlined we and rates, flow and times flow water optimized schedules, feeding developed we sections, out grow- and nursery the In nursery. the to transfer before as maintained are such embryos/larvae which food, live as well as food dry and powdered paramecia, size-specific and age- use we ZIRC, the At to theturnoveroffishgenerationsatResource Center. growing zebrafish at ZIRC. Thus, the nursery and larval grow-out sections contribute significantly optimal and cost-efficient husbandry methods, we improved the efficiency, speed, and success of Our goal is to optimize the throughput, survival, growth rate, and health of zebrafish. By adapting Zebrafish International Resource Center, University ofOregon, Eugene, USA C. Barton The ZIRCNurseryandGrow-Out Section 75 . We also incorporated the use of a modified E2 embryo medium in medium embryo E2 modified a of use the incorporated also We . artemia Husbandry Workshop basis ofthehumandisorder. suf zebrafish, the eggin phenotype willdefects an importantswimming tool to understandobserved the cellularnot and molecular have we Although muscles. leg the of paralysis in resulting endocytosis in other tissues. In fact, mutations in the human homolog cause hereditary eggs. paraplegia In addition, we find allele. mutant Interestingly, our in deleted domain specific vertebrate a detect and genomes vertebrate and trafficking vesicle the in with consistent observed endosomes defect to localize to predicted are domains FYVE that shows mutation the of cloning Positional during endocytosis. trafficking vesicle controls Soufflé that and compartment lysosomal hydrolyzing the in arrives never protein yolk oocytes mutant in that Togethersuggest opaque. results stay these to in contrast, In cleaved. proteolytically is protein yolk because transparent, become eggs type wild suf mutants and in yolk does accumulate not form vesicles crystalline arrays small in yolk oocytes, globules. During wild-type electronoocyte maturation to performed contrast In we oocytes. mutant oogenesis, on during microscopy Soufflé of role the understand better To meiosis. in of nucleus the cytoplasm, for zebrafish mutations causing defects during oogenesis. Unlike wild-type eggs with transparent (Suf). Soufflé called endocytosis of regulator novel a Weidentified be to tissue culture,whichdiscoveredmanyofthekeymoleculescontrollingthiscellularprocess. need they oocyte, the in expressed not imported by clathrin-mediated endocytosis. Endocytosis has been intensely are studied in yeast and proteins yolk Since developed. has system In egg laying animals yolk provides essential nutrition for the embryo until a functional digestive Marine Science(CSIC), Barcelona, Spain 1 R. Dosch The soufflégeneisrequiredfor vesicle trafficking duringoogenesis Gern cellsandmaternaldeterminants et o Zooy n nml ilg, nvriy f eea Switzerland; Geneva, of University Biology, Animal and Zoology of Dept. oocytes small vesicles persist and the uncleaved yolk protein causes the egg cytoplasm egg the causes protein yolk uncleaved the and persist vesicles small oocytes suf 1 , A. Stein suf oocytes stay opaque similar to immature oocytes. However, our results show that is also present in mammalian genomes, which do not deposit yolk into their into yolk deposit not do which genomes, mammalian in present also is suf oocytes progresses through all steps of oocyte maturation excluding a defect a excluding maturation oocyte of steps all through progresses oocytes suf 1 , F. Bontems uat. hlgntc tde dsoe Sf ooos n invertebrate in homologs Suf discover studies Phylogenetic mutants. suf suf expression outside the zebrafish germline indicating that it controls 1 , J.Cerda 2 , J.Lyautey encodes a zinc finger FYVE domain protein. domain FYVE finger zinc a encodes suf 1 was isolated in a screen a in isolated was Suf 76 2 RA Isiue of Institute IRTA, 147

Posters Posters T. Müller transplantation protocol provides a means to study cellular and molecular aspects of oocyte of aspects molecular and development inthezebrafish. cellular study to means a provides protocol transplantation oocyte stage early The activities. gene maternal with interfering for tools developing to way the of microinjection simple into paves and oocytes develop stage that early in expression gene reporter RNAin with results follicles I-II they stage show weeks we Moreover,3 offspring. in viable and produce weeks and 2 eggs in IV mature stage to develop can oocytes I stage donor of proof provide where we mothers recipient into Here zebrafish of follicles I-II developed. stage transplanting for be protocol principle to needs methods oocytes novel in factors Therefore, maternal tools. manipulate genetic to reverse and forward conventional by key products of gene maternal regulator with interfering of difficulty and essential technical the embryonic due an estimate to to hard contribute beis development effects larval to maternal which continues to contribution but extent MBT, The maternal processes. at However, developmental stop fishes. not including does models embryogenesis animal to blastula many mid the in until (MBT) exclusively almost transition embryogenesis early drive products gene Maternal Birmingham, UK 2 1 148 F. Intraovarian transplantation ofstageI-IIfolliclesresultsinviablezebrafish embryos Germany. Institute of Toxicology and Genetics, Forschungszentrum Karlsruhe,Eggenstein-Leopoldshafen,ForschungszentrumGenetics,Toxicology of and Institute Hungary.University,Gödöllő, Istvn Szent Environme, of Institute Culture, Fish of Department Müller 77 1 , 3 2, , B.Urbanyi 3 eatet f eia ad oeua Gntc, eia Sho, nvriy of University School, Medical Genetics, Molecular and Medical of Department Z Csenki Z. , 1 , L.Varadi 1 A Zaucker A. , 1

2,3 BY Kovacs B.Y. , Gern cellsandmaternaldeterminants 3 Y Hadzhiev Y. , 4 A Hegyi A. , 1 KK Lefler K.K. , 1 , reminiscent of a mouse most The cRNA. Irxl1 of coinjection by is phenotype archThis and cartilages. muscles craniofacial the in observed were defects rescued severe partially be can phenotype The days. 7 to 5 of knockdown morpholino Antisense arches. pharyngeal two first the and brain the in mainly expression Irxl1 Whole-mount tissues. adult in expressed broadly were to vertebrate orthologs. Transcription of was z-Irxl1 detected from 18 hpf to 5 dpf. Both isoforms identical % 100 homeodomain a encodes and exons 7 contains It 12. chromosome on located myoD-dependent pathways. by mediated be may development arch in function its and morphogenesis, arch and muscle brain, of regulator of inhibition with of construct somites luciferase promoter-driven the in increased was expression in decreased was of activity promoter the up-regulates embryos zebrafish into the of Irxl1 homeobox genes that is most-closely related to the development. Homeobox genes are transcription factors that play important roles in the regulation of embryonic ofBiomedical Sciences,Department ChungShanMedicalUniversity, Taichung, Taiwan H. Pan, H.N.Chuang, H.Y. Cheng,C.C.Wu in zebrafish development arch pharyngeal and muscle brain, in involved gene homeobox novel a is Irxl1 Early developmentandgastrulation gene and obtained the full-length cDNA of two splicing variants. Zebrafish variants. splicing two of cDNA full-length the obtained and gene ee otis eea cness e2 idn sts ad oijcin f Mef2ca of co-injection and sites, binding Mef2 consensus several contains gene Irxl1 promoter activity by Irxl1. These results suggest that Irxl1 is an important an is Irxl1 that suggest results These Irxl1. by activity promoter myoD Iroquois homeobox-like 1 ( homeobox-like Iroquois resulted in deformed head and jaw in the larvae, which only survived for survived only which larvae, the in jaw and head deformed in resulted Irxl1 Mroe, rl protein Irxl1 Moreover, Mef2ca. mutants and morpholino-knockdown morphants. Meanwhile, morphants. morpholino-knockdown and mutants Mef2ca Twirler mutation, to which the ) is a novel member of the TALE superfamily of superfamily TALE the of member novel a is ) Irxl1 into zebrafish embryos revealed adramatic revealed embryos zebrafish into Irxl1 kokon mro. Co-injecting embryos. -knockdown Irxl1 Iroquois class. We have identified the zebrafish a rglt msl dvlpet through development muscle regulate may gene is linked. The Irxl1 promoter region hybridization analysis revealed analysis hybridization situ in . In addition, In . Irxl1 78 Irxl1 Irxl1 gene is gene Irxl1 expression myoD myoD 149

Posters Posters 150 K.M. Hsiao Functional studiesofmuscleblindproteinsinearlyzebrafish development 1 MyoD-dependent pathway. for early fish development and that myoD reduction of MHC expression. Dual-luciferase assay further reveals that zmbnl3 down-regulates to inhibit cell differentiation, as evidenced by lack of cell fusion, change of splicing pattern and cells, C2C12 into introduced When somites. short and axes body crooked with of microinjection contrary, the are expansion, RNA CUG with cells in altered in the morphant misregulated embryos. Knockdown of are that pre-mRNAs several of patterns are defective in hatching and swimming morphants behavior.these addition, bladder.In In swim consistent and with heart the arches, phenotypes, pharyngeal the otoliths, splicing eyes, brain, ubiquitous rather than specific. Knockdown of branchial arches, midbrain hindbrain boundary and swim bladder. Expression of zmbnl3 is more while muscle, liver,and lens, vesicle, in otic situ in Whole-mount stage. pharyngula late until contrast, In expressed. zygotically and maternally both are embryogenesis, During tissues. adult most in expressed broadly are genes three the of splicing eeomn, e lnd the cloned we development, to RNA-mediated pathogenesis. To or investigate the functions of Mbnl proteins in early vertebrate inclusion promotes leads human in activity each MBNL of Misregulation mammalian, genes. different on in exons specific of identified exclusion been have 3) and 2, (Mbnl1, Three domain. paralogs finger zinc CCCH conserved a through RNA to bind They tissue-specific splicing. of alternative regulation the in participate that proteins of family a is (MBNL) Muscleblind 3 Department ofBiomedical Sciences,Department ChungShanMedicalUniversity, Taichung, Taiwan University,Chia-Yi,Cheng Chung National Science, Life of Department 79 promoter activity in fish embryos. These data indicate that 1 , L.C.Tu 2 primary transcripts gives rise to at least 13 protein isoforms. These isoforms. protein 13 least at to rise gives transcripts primary zmbnl , C.W. Lin genes in zebrafish (zmbnl1, zebrafish in genes muscleblind 3 andH.Pan cRNA into the embryos resulted in defective embryos defective in resulted embryos the into cRNA zmbnl3 zmbnl3 may interfere with muscle differentiation through the 3 is expressed in lens, olfactory epithelium, olfactory lens, in expressed is zmbnl2 zmbnl1 and Early developmentandgastrulation zmbnl3 does not result in overt phenotype. On hybridization reveals that reveals hybridization zmbnl2 transcripts are not detected not are transcripts zmbnl3 zmbnl1 and zmbnl2 are crucial results in malformation of the 2 and 2 Institute of Medicine, of Institute and zmbnl1 is expressed is zmbnl1 is able is zmbnl3 . Alternative 3). zmbnl2

1 K. Goudevenou signalling pathway Wnt non-canonical the in factor,activatesWnt5 exchange nucleotide guanine novel a Def6, Early developmentandgastrulation it playsinregulatingCEmovementduringgastrulation. role the elucidate further to point starting a provide and pathway signalling Wnt non-canonical the in Wnt5 of downstream acts def6 that show far so results Our phenotype. knockdown def6 severe the rescued RhoApartially tested active def6.Constitutively of further downstream RhoAacts we whether RhoA on converges signalling Wnt non-canonical Since pathways. parallel synergisticallyand Wnt11pathway with same the in Wnt5 of downstream or same the either in functions def6 that Togethersuggest RNA. results def6 these by rescued largely was phenotype synergism, def6 RNA did not rescue the Wnt11 knockdown. In contrast, the Wnt5-MO-induced suggested Wnt11MOs and def6 of co-injection Although RNA. def6 with phenotypes induced MO- Wnt5 Wnt11and the rescue to attempted we Wnt5 Wnt11and of downstream functions the that indicating MO-induced defects phenotype were a def6-Morpholino-mediated direct result of the specific def6 rescue knockdown. to Tosufficient determine whetherwas def6 def6 protein of down knock that show def6 GFP-tagged of Overexpression we specification. fate cell not but movements, cell CE Here, affected unknown. remain movement cell and polarity cell However,cells. shape, mammalian cell in regulating F-actin in with has localizes Def6 role the co- also and Cdc42 and Rac Rho, for of GDP upstream acts bound Def6 of that GTP.shown exchange previously Wehave the through GTPases Rho activating GEF of type novel a is Def6 GTP exchangereaction. GDP- the mediate directly can Daam-1 nor Dishevelled neither this as unknown, remains in pathway involved GEF the of identity nucleotide the However, guanine activation. their require for (GEFs) Rac factors and exchange Rho forms. GTP-bound active and GDP-bound inactive Rho, Rac and Cdc42, act as molecular switches and regulate Rac the leading to cytoskeleton the reorganization via of the cycling actin cytoskeleton. between The Rho family of GTPases including through their Frizzled receptor, Dishevelled, and Daam1 to activate the small GTPases Rho and signal Wnt11Wnt5 glycoproteins and secreted two The fate. cell affecting without movements the CE defective to wnt11 lead the genes in (Pipetail) mutations wnt5 zebrafish, and In (Silberblick) mediates that pathway (PCP) polarity cell . establishment ofcellpolarityintheplaneepitheliaDrosophila planar the to similar pathway, Wnt canonical non- the by regulated is VertebrateCE axis. embryonic anterior-posteriorforming the and of (convergence)(extension) lengthening narrowing mediolateral the to leading another one with intercalateand midline dorsal the toward move cells where (CE), extension convergenceand is germ layers: endoderm, mesoderm and ectoderm. One of the major cell movements taking place Gastrulation is driven by coordinated cell movements that form the embryo proper into the three Nottingham, UnitedKingdom nttt o Gntc, col f Biology, of School Genetics, of Institute 1 , P. Martin 1 , Y.J. Yeh 1 , P. Jones 2 col f imdcl cecs Te nvriy of University The Sciences, Biomedical of School 2 , F. Sablitzky 1

80 151

Posters Posters 152 medaka 512-cell-stage to presence ofactivatedSTAT3. 64-cell-stage nuclear on spanning depending is which development STAT3 medaka in process specific a for of argues embryos import-wave the Furthermore, mouse-system istheonlyvertebratestemcellsystemdependingonactiveSTAT3.the Hence, medaka. in status stem-cell for necessary STAT3not activated is that indicate data STAT3that demonstrated Consequently,cells. stem embryonic medaka these in activated not is Conclusions: and Discussion after 512-cell-stage. STAT3-levelsnuclear of increase No 64-cell-stage. at peak or 64-cell-stage before detected was STAT3a with 512-cell-stage and 64-cell-stage between nucleus the into import wave-like a has that STAT3 is inactive in all cells. Investigations on embryos before 1024-cell-stage revealed that demonstrated blastula-stage of embryos medaka in Analyses MESI. in localization inactive thus Results: via immunofluorescence. investigated STAT3-activity was factors. fish growth medaka of embryos early and MESI-cells in fish (Oryzias latipes) and remains undifferentiated in cell culture in presence of a complex mix of Material and Methods: stem cellsofmedakafish(MES)andduringearlyembryonicdevelopmentmedaka. status. In this work, we analyzed the localization and thereby the activity of STAT3 in embryonic neither necessary nor sufficient to keep human embryonic stem (hES) cells in the undifferentiated and consequently imported into the nucleus. In contrast to mES-cells, activated nuclear STAT3 is (LIF) factor STAT3inhibitory factor leukaemia transcription the latent by The activated status. is „stemness“- the in cells mES keep STAT3,to proteins, needed transcription is of activators and achieved. In murine embryonic stem (mES) cells, one member of the family of signal transducers is cells stem in differentiation spontaneous of prevention the and maintained is self-renewal for Introduction: Physiological I, Chemistry Biozentrum, University of Wuerzburg, Germany M. Kraussling, T. U.Wagner, M.Schartl latipes Oryzias of development early during and cells stem STAT3-activityembryonic of in Analysis 81 Ectopic expression of an eGFP-tagged STAT3 resulted in dominantly cytoplasmic and cytoplasmic dominantly in STAT3resulted eGFP-tagged an of expression Ectopic An important aspect in stem cell research is the understanding of how the potential The embryonic stem cell line MESI was obtained from blastulae of medaka h asy o MS cls n batl-tgd eaa embryos medaka blastula-staged and cells MESI on assays The Early developmentandgastrulation to thedifferentfamiliesfromsameorderCypriniformes. receptor. belongs which species studied two in defects developmental induced in difference significant a 5HT1a zebrafish the demonstrated drugs serotonin-related to of screening Pharmacological preparations. wholemount corresponds which kDa 44 of Utilizing the size same antibody we detected with a distinct punctate band signal on the surface a of blastomers in revealed receptor 5HT1a human product against the antibody 5-HIAA, with analysis and blot Western5-HT degradation. both serotonin of of concentration the increase precursor 5-HT with eggs fossilis). loach(Misgurnus weather two andEurasian 5-HT expression started in ofzigote and continued in the blastoderm. Incubation of dichorioneted blastula) rerio) to (Brachydanio zigote zebrafish (from fishes, development teleost early the during receptors serotonin and 5-HT of distribution the describe to microscopy confocal scanning laser and biochemistry used We ofmorphogenetic themechanisms study to effects of5-HT. model afavorable embryos zebrafish the makes Takenembryo. developing a in facts techniques this molecular altogether and pharmacological cytological, of range wide a use to opportunity an provides and details, great in described been well are system serotonergic the known in larval and of adult nervous system of zebrafish. properties Normal development of teleost pharmacological fishes has and physiological However, No data on the serotonergic system at the fish development before appearance of neurons exist. still purelyunderstood. are processes morphogenetic in neurotransmitters of functions the data, existing all of Despite development. early on impact an have system serotonergic on targeted that drugs neurotropic Thus, stages. as 5-HT receptors have developmental been found in early various animals starting from very oocyte stage. It is the known that from serotonin (5-HT), enzymes necessary active for 5-HT synthesis, transportation and degradation, is the as well However, example, tissues. for complex and system, cells serotonergic differentiated contains which organisms adult in of or Most culture cell years. in either neurotransmitters 100 of role past physiological the on the focused were studies over these studied extensively were systems neurotransmitter Various Institute ofDevelopmental Biology, Russian Academy ofSciences, Moscow, Russia of E. G.Ivashkin, E.Voronezhskaya stages prenervous early the at drugs teleost fishdevelopment neurotropic of effects the and system Seretonergic Early developmentandgastrulation 82 153

Posters Posters 154 is applicable formutagenesisinzebrafishthoughitsefficiencyislow.system trap gene Tol2transposon the that indicates experience Our detail. in studied being Weeyes. and are functions their and lines mutant these in genes interrupted the identified have extension sac yolk reduced pigmentation, reduced edema, hydrocephalus, of presence the by fertilization post hours 28 at recognized be can mutants YT101 dpf. 5 at die then bladder,and mutants YT068 exhibit small eyes, a reduced brain, lack of jaws, branchial arches and swimming insertions have been identified from 121 F2 families. Beginning at 2 days post fertilization (dpf), transposon the screening, to linked tightly are that mutations lethal recessive Four pilot identified. were fashion temporal and the spatial in In GFP expressing mated. families F1 183 and and screened raised were fish were injected 4900 embryos injected The stage. one-cell at eggs and 5’ transposon vector reduced and the transposase mRNA the synthesized in vitro contains were Tol2,co-injected into which fertilized of TSBGS, ends vector,3’ trap gene Tol2-based a constructed We mutagen. were insertional efficient an is development system transposon The screening. zebrafish mutagenesis by identified for essential genes Many out. carried been have screens genetic insertional and chemical of strategies the diseases, and development in involved genes identify development. To and genetics vertebrate studying for organism model powerful a is Zebrafish ofBiological SciencesandBiotechnology,Department Tsinghua University, Beijing, China S. Yang, J.Tong, Y. Han,X.Zheng, P. Xu, A. Meng Mutagenesis inZebrafish by Transposon Gene Trap System 83 piig cetr GP eotr ee n te V0 oy sga. The signal. polyA SV40 the and gene reporter GFP acceptor, splicing bcl2 Early developmentandgastrulation the new the α-cat in locally regulating actin and membrane dynamics at sites of cell-cell adhesion. However, of in increase local a that proposed been concentration during clustering of the cadherin/catenin complex could regulate local formation has it adhesion; cell-cell of sites at cadherin to 2005). al., dynamic equilibrium between et monomer Yamadaand dimer 2005; pools in al, the cytoplasm and et monomer bound (Drees complex cadherin/beta-catenin the with dimeric while actin, not but monomeric which in protein quaternary complex with cadherin, that however,beta-catenin and showed, actin. Instead model this of test direct A complex. beta-catenin cytoskeleton, actin the to bound is the AJ with the which in complex stable a depicts AJ the of model defects. injected embryos, these embryos complete somitogenesis. However, they display morphological mitochondria mislocalization of caused delayed or arrested in epiboly and the embryos do not complete somitogenenis. Inducing for the cytosol and plasma membrane pools. Our preliminary results shows that the morpholino diverting the the dynamically perturb to is approach the mitochondria or to the plasma membrane (in a AJs independent manner). The rational of this mislocalized to able the are that constructs of number a approach, new morpholino the the with verify together to adhesion, cell for system model a cytoplasmic their through bind that and p120 to proteins domain adhesion cell transmembrane family, cadherin between balance the the of members cancer.comprise and as AJs The healing wound embryogenesis, during transition of dynamic organization epithelial-mesenchymal is and migration the cell organizationin roles important and plays AJ disassembly and polarity.assembly adhesion, cell cell-cell and for domains essential membrane are (AJs) junctions Adherense The University,Stanford USA A. Schepis, W. S.Talbot andJ.W. Nelson Allosteric Controlof Alpha-Catenin inCell Adhesion andMigration Early developmentandgastrulation α-cat dimers in the cytoplasm. These results point towards a dynamic, rather than static role of -cat interfering with his allosteric regulation. These constructs are able to direct to able are constructs These regulation. allosteric his with interfering α-cat -cat’s properties were shown by shown were properties α-cat’s -cat providing the molecular link between the actin cytoskeleton and the cadherin/ the and cytoskeleton actin the between link molecular the providing α-cat α-cat to the mitochondria, we can lower the concentration of the protein available -catenin; and β-catenin; -cat binds to and bundles actin filaments but does not interact not does but filaments actin bundles and to binds α-cat -cat is able to form a ternary complex with cadherin/β-catenin, with complex ternary a form to able is α-cat α-cat causes delay in epiboly, but in contrast to the morpholino -catenin in turn binds to binds turn in β-catenin -cat concentration of the different pools. For example, For pools. different the of concentration α-cat study. We chose the zebrafish gastrulation, zebrafish the chose Westudy. vitro in ct properties α-cat -cat). The textbook The (α-cat). α-catenin α-cat behaves as an allosteric 84 W ae using, are We vivo . in -cat cannot form a form cannot α-cat -cat is in is α-cat -cat to α-cat α-cat 155

Posters Posters Pou5f1 indorsal-ventralpatterning. our data provide embryological and molecular evidence for the direct activation of the of activation direct the for evidence molecular and embryological provide data our conserved and effects hardly it while (CHX), cycloheximide of presence the in epiboly,even 40% by a of quantities larger of that zygotic maternal produce to mate fish These mRNAinjection. zebrafish, In populations. cell ohne grenzen ( spg) shows neural plate pattering defects. We pluripotent rescued the embryonic for zygotic The expression. zygotic and required maternal both has pou5f1/pou2 is mice in Pou5f1/Oct4 University ofFreiburg, ofDevelopmental Department Biology, Freiburg, Germany B. Wendik, K.Lunde,B.Polok,HG.Belting,W. DrieverandD.Onichtchouk expression for overexpressing embryos wild-type in and mutants MZspg 156 of Overexpression MZspg. including factors, ventral other several of expression that revealed analysis MZspg mutants, like wild-type embryos, can be further dorsalized in response of that and epiboly, via of activation transcriptional by patterning dorsoventral to contributes Pou5f1/Oct4 Pou5f1-Endorsalize. can protein fusion repressor 85 in the in pou5f1 and bmp2b acts as a transcriptional activator during dorsal-ventral patterning. Overexpression patterning. dorsal-ventral during activator transcriptional a as acts pou5f1 expression in the presence of CHX. In silico analysis of the of analysis silico In CHX. of presence the in expression bmp4 binding sites which could be confirmed by gel shift assay. Taken together, assay. Taken shift gel by confirmed be could which sites binding Pou5f1 expression assays, we see that see we assays, expression fgf8 pathway leading to ventral tissue specification. In support of this idea, this of support In specification. tissue ventral to leading pathway bmp can dorsalize wild-type embryos prior to 30% epiboly. Notably,epiboly. 30% to prior embryos wild-type dorsalize can Pou5f1-En can ventralize wild-type embryos, while overexpression of overexpression while embryos, wild-type ventralize Pou5f1-VP16can Pou5f1-VP16activates strongly Early developmentandgastrulation mutants are dorsalized prior to 30% to prior dorsalized are mutants MZspg can rescue dorsalization both in both dorsalization rescue can caBMPR1a vox . Thus, we propose a role a propose we Thus, Pou5f1-En. expression in wild-type embryos wild-type in expression spg ( ) mutants, indicating mutants, MZspg) mutant spiel mutant pou5f1/pou2 spg mutants by vox vox promoter reveals promoter fgf8. Microarray , is affected in affected is , bmp2b pou5f1 vox vox by

by the anterior-posterior axis of zebrafish embryo showed an asymmetrical development, the development, of expression asymmetrical an showed embryo zebrafish of axis anterior-posterior the by analyzing and microinjection the expression of by zebrafish stage two-cell the of cell one in mRNA expressing Flounder of function The vesicle etc. pineal and cells, gland. crest neural trunk and cranial placodes, otic post somites, cells, intestine. crest neural and brain cells, progenitor kidney somites, embryogenesis, of stage early the During domain. binding flounder The pattern andfunctioninregulatingmusclewereanalyzed. involved inmuscleformationofflounder, FoxD familygeneswereisolatedandtheirexpression muscle development and maturation. To further clarify the function of regulatory factors that are region, flounder has been Asian cultured the for many in years,fish butimportant thereeconomically is littlean physiology. informationAs aboutand itspatterning skeletal embryonic roles early important in highly play family a this contained of genes them shown of been had All It species. domain. binding metazoan DNA conserved of variety a in identified are family this of members 100 than fly, more fruit from discovered was gene (Fox) Box Forkhead first the Since Qingdao, ShanDong, P.R.China Experimental Marine Biological Laboratory, Institute of Oceanology,Chinese Academy of Science, (paralichthysX. Tan flounder ,Y. Zhang,PJ. Y. from Xu (1,3,5) foxd analysis olivaceu) function and pattern expression Cloning, Early developmentandgastrulation support ofK.C.Wong EducationFoundation,HongKong. Oceanology,of Institute Chinese the acknowledges gratefully author The Sciences. of Academy Program, Biology,Marine (973 Experimental of Laboratory Key the and 2006AA10AA402) China No. Program, (863 of Program Research Basic No.2004CB117402)，the National National High (Y2008E12),the Technology Tan Research Xungang and Development to Program Province，P.R.Chinaof Shandong of China Foundation Science Natural the by supported was work This FoxD1 orFoxD5.So, theregulationofFoxD1,3,5 wasdifferentfromeachother. over-expressed Flounder over-expressed Flounder zebrafish engogenous the analyze to performed of So, enhanced. affected; not expression in the somites on one side was enhanced, but the expression in the adaxial cells was adaxial cells and somites on one side was reduced. When reduced. was side one on somites and cells adaxial affected; not was cells adaxial the in expression the but reduced, was side one on somites the in expression MyoD expressed, over was mRNA MyoDand was expressed in several tissues including somites, tail bud, forebrain and otic and forebrain bud, tail somites, including tissues several in expressed was FoxD5 expression in the somites and presomitic mesoderm on one side was also was side one on mesoderm presomitic and somites the in expression Myf5 contain only one exon and a conserved winged helix DNA helix winged conserved a and exon one FoxD1only ，FoxD3contain ，FoxD5 and MyoD . ToMyf5. of mechanism regulation the understand further might regulate the muscle development by regulating the expression the regulating by development muscle FoxD1,3,the regulate might 5 Myf5 and FoxD1 or FoxD3 can inhibit the expression of zebrafish endogenous in the paraxial mesoderm was inhibited. When Flounder When inhibited. was mesoderm paraxial the in Myf5 FoxD1,3,5 MyoD. After flounder FoxD5 can not inhibit the expression of zebrafish endogenous n uce eeomn ws odce truh over- through conducted was development muscle on FoxD1,3,5RT-PCRexpression. the that showed expression in the presomitic mesoderm, presomitic the in expression Myf5 FoxD3 overexpressed, the two sides separated a epesd n premigratory in expressed was FoxD3 was mainly expressed in the in expressed mainly FoxD1was FoxD5 was over expressed over was 86 FoxD1,3,5RT-PCR, was FoxD3,while FoxD1 MyoD 157

Posters Posters 158 shape cell controlling be could JNK changes andmotilityunderlyingmophogeneticmovementsinearlyzebrafishembryogenesis. that propose We movements. circling display and body a convergence and extension phenotype with a shorter and broader body.both cell monitor adhesion and cytoeskeleton. They WeEmbryos that overcomealso the layers. DCs have epiboly phenotype show a different for curled markers with sections the fixed in and videomicroscopy between confocal by behaviour adhesion cell in changes cell controls JNK that suggests This toanepiboly rate. leads JNK while epibolic expansion ofzebrafish of the superficial EVL and the YSL nuclei seems to progress Knock-down at a normal embryo. phenotype, in zebrafish which migration of the majority the of deep cells is affected in in the morphant embryos, extension and signalling pathway as a potential candidate andcell to regulate the processes of epiboly and cell polarity convergence the yolk syncitial layer (YSL) both could be directed by many signalling directing mechanisms.We propose JNK and (DCs) mechanism layer cells deep the (EVL), specific layer enveloping the between interactions The suggests migration. layers cell diferent the of epiboly,sheets. At epithelial of fusion and movement expansion coordinated the the in implied The zebrafish embryo is an ideal model system to study the molecular and cellular mechanisms Barcelona, CSIC, Barcelona de Molecular Biologia de Spain Instituto Biology, Molecular and Cell M. MarsalandE.Martin-Blanco The roleofjnkinzebrafish earlymorphogenesis 87 Early developmentandgastrulation 1 oe f h wtpp aha cmoet rlbt/ag2 n el eair underlying behaviors cell in trilobite/vangl2 I. Roszko component pathway wnt/pcp convergence andextensionmovements the of Role Early developmentandgastrulation akd n hgl mdoaeal eogtd ad irt fs ad ln srih trajectories straight along and fast migrate towards dorsal. But and elongated, mediolaterally highly and packed tightly become cells embryos, wild-type in proceeds, gastrulation As normal. is migration cells in cellpolarizationduringC&E. components PCP these for roles distinct suggest we studies these on Based polarized. properly MTOC polarization for in cells essential engaged in be C&E at to late gastrulation stages, appear in Knypek/Glypican4, and Dvl Whereas C&E. in engaged cells in position (MTOC) centers organizing microtubule of position the analyzing by cytoskeleton, microtubule the of polarization Tri/Vangl2and between relationship the investigated Vangl2-GFPalso We a Using localization. Tri/Vangl2 perform asymmetric we construct involve not mediolateral that does show We polarization localization. cell asymmetric preferential its in results membrane the et (Yin in gastrulation lost is during distribution This mesoderm 2008). in al., localized thatPrickle asymmetrically are datashowed (Dvl) Dishevelled Recent and behavior. cell of during modification time with specific correlating a at gastrulation, membrane cell the to recruited Tri/Vangl2is endogenous that show Wemovements. C&E during behaviors and morphology cell in transition this Wecharacterize in stage, this At migration. directed by midline dorsal the towards slowly converge to begin and packed loosely are cells mesodermal lateral midgastrulation, At PCP component,Trilobite/Vangl2 (Tri) inthisprocess. cell polarity in vertebrates remains a challenging open question. We investigate the role of a core body.embryonic nascent the elongate PCPthe whereby mechanism the But, regulates pathway behaviors cell polarized and narrow which movements, (C&E) extension convergenceand during for cells mesenchymal of necessary are components PCP is vertebrates, architecture in tissue However, such absent. cells mesenchymal In domains. membrane specific to localization asymmetric characteristic a PCPexhibit and components junctions pathway tight by connected essential role in the establishment of planar polarity in The Wnt/Planar Cell epithelial Polarity (Wnt/PCP) tissues. pathway was In discovered in these Drosophila where epithelia, it plays cells an are Medicine/Division ofGeneticMedicine, Vanderbilt University MedicalCenter, Nashville, USA eatet f ilgcl cecs Vnebl Uiest, ahil, USA, Nashville, University, Vanderbilt Sciences, Biological of Department 1 , J.R.Jessen tri 2 mutant cells fail to elongate and their movement is slow and less effective. , D.Sepich tde t aaye h dnmc eair f Tri/Vangl2 protein. of behavior dynamic the analyze to studies vivo in 1 , Y. Liu mutants. We asked whether Tri/Vangl2 recruitment at Tri/Vangl2recruitment whether asked We mutants. tri 1 , L.Solnica-Krezel 1 MZtri mutants the MTOCs, are tri 88 mutants, the mesodermal the mutants, 2 eatet of Department 159

Posters Posters f enyvna Piaepi, USA Philadelphia, Pennsylvania, of 4 1 160 C. M.Cruz Functional Analysis oftheeve1 GeneDuringZebrafish Neural Development important bothinposteriorisationaswelltheinductionofneuraltissuetrunkandtail. that found we addition, In signal. found that embryos and have uncovered an important role for we have performed loss-of-function and early marker analyses in wild type sofar and species vertebrate all overexpression experiments have in revealed its role identified in tail development been in both since fish and frog.studied. In vertebrates, have it Here, is expressed in the posterior during early embryonic development, and homologues eve polarity, A/P in fly fruit the in characterised Originally is a zebrafish homologue of the of homologue zebrafish a is Eve1 Dept. Anatomy andDevelopmental Biology, University College London, UnitedKingdom Exeter,Biosciences,of of University School Devon, Kingdom,United 89 eve1 activates the expression of posterior genes, such as 1 , S.Maegawa 2 , E.S.Weinberg possesses neural inducing activity and may therefore be therefore may and activity inducing eve1neural possesses 3 a. oeua Gntc, IH, I, ehsa USA, Bethesda, NIH, NICHD, Genetics, Molecular Lab. even-skipped ( wee t a a rca role crucial a has it where , melanogaster Drosophila 2 , I.B.Dawid Early developmentandgastrulation eve1 in both trunk and tail development. We eve 3 , S.Wilson ) family of transcriptional repressors. transcriptional of family ) hoxb1b, via the retinoic acid 4 , T. Kudoh 2 Dept.Biology. University 1

ichabod mutant P. Schu 1 D.Zizioli Characterization ofthe AP-1 μ1-adaptininzebrafish (Daniorerio) Early developmentandgastrulation nvriy f aln, Italy; Mailand, of University ap1m1 is located on chromosome 2 while Zebrafish genes. mouse and human the to compared then and Browser Genome ENSEMBL the orthologs were determined by performing BlastN alignments of different AP-1 μ1 cDNAs against mammalian μ1-adaptins. The genomic structure and the exon-intron boundaries of the zebrafish corrisponding the to identity 83-85% share sequences identified respectively.two subunit, The μ1A expressed a ubiquitously the as and μ1B tissue-specific ) rerio the encoding sequences analysis (Danio two database revealed silico In zebrafish subunits. μ1B and used μ1A of Wefunction the explore development. to model vertebrate embryonic mammalian for essential is polarized the and μ1A expressed epithelia-specific μ1B.ubiquitously Mice “knock out” for the subunits the as exists subunit μ1-adaptin mammalia In vertebrate.We are currently running “knock down” experiments in order to elucidate also the role playedbyμ1-A adaptinduringzebrafishdevelopment. also elucidate to order in experiments down” “knock running currently are vertebrate.We of development organs in μ1B-adaptin by played role essential the of demonstration first the is severe during the late stages of more development and thereafter development became ceased at 7-8 dpf. mobility,that This reduced a showed derivation. morphants endodermal the onward of hpf 24 tissues Moreover,from and liver intestine, of formation defective by characterized of μ1B expression by using Depletion “translation-blocking morpholinos” (Mos formation. vesicle AP-1 restore and deficiency μ1 mouse the complement could ap1m1 and such as gut, pharingeal endoderm, liver and pancreas. When the coding sequences of zebrafish while notochord, the in The expected. as that revealead analysis sections Histological derivation endodermal of organs in present mount whole was transcript μ1B the while ubiquitously,present was μ1Atranscript The hpf. 72 to somitogenesis late from observed were levels high whereas embryogenesis, of stages first the at detectable already are transcripts μ1B and μ1A that revealed stages developmental different on RT-PCRanalysis The introns. 10 and the first one is organized in 10 exons and 9 introns and the second one is organized in 11 exons vesicles by subunits: mediated four of is consists AP-1 endosomes AP-1. complex adaptor-protein and the network by formed trans-Golgi the between transport Protein Germany The intron-exon structure of zebrafish e. f imdcl cec ad itcnlg Uiest o Beca Italy; Brescia, of University Biotechnology and Science Biomedical of Dep. 3 1 , M. Guarienti M. , ap1m2 hybridization confirmed the results obtained by subunits. by RT-PCR both obtained for results the confirmed hybridization situ in were transfected in μ1-deficient mouse embryonic fibroblasts, both subunits is expressed only in organs of endodermal derivation such as such derivation endodermal of organs in only expressed is ap1m2 1 , R. Bresciani R. , 3 etr o Bohmsr, er Ags Uiest, Gottingen, University, August Georg Biochemestry, for Center ap1m1 1 , E. Monti E. , ap1m2 is expressed ubiquitously and in particular in and ubiquitously expressed is ap1m1 and 1 ap1m2 , G. Gariano G. , is not yet located on a specific chromosome. γ1 or μ1A revealed that AP-1 complex is similar to human and mouse genes: 1 , C. TobiaC. , ap1m2) gave rise to embryos 90 1 , A. Preti , A. 2 e.f Biology, Dep.of γ1, 1 1, μ1, β1, , F., Cotelli 161 σ1. 2 ,

Posters Posters a marker of nervous tissues and eyes development. A significant reduction of reduction significant A development. eyes and tissues nervous of marker a help us to elucidate the function of role played specific by a indicating further controls, standard to compared morphants ap1γ2 in detected was of intracellularvesiculartraffic. nvriy f aln, Italy; Mailand, of University 1 162 M. Guarienti AP-1) inzebrafish (Daniorerio) novel a of characterization functional and Identification 2 morphants, using morphants, ap1γ2 on hybridization situ in whole-mount performed We (7-8 dpf)andthentheydye. not able to leave from the chorion; they maintain this immobility till later stages of development are they since immobility phenotype severe showed morphants the addition In system. nervous the that showed results obtained the named subunit, was protein, cloned. related adaptor novel a recently and characterized well is subunit formation endocytic pathway and consists of which four subunits: vesicles, clathrin-mediated involves requires pathways an adaptor protein endocytic complex (AP). The and AP-1 complex mediates transport exocytic from TGN to the In eukaryotic cells, transport of proteins between the different membrane compartments through Germany xrsin lo n ri ad ys Sai-eprl xrsin nlss y hl-on in whole-mount morpholino (Mos by analysis expression translation-blocking the RT-PCR. by used obtained We results the Spatio-temporal confirmed hybridization situ eyes. and brain in also expression transcript showed organs adult in analysis same The hpf. days 5 to cells) (1-2 stages early from exon-intron RT-PCR the analysis on and total embryo RNA structure showed that the genomic exons and20introns.Thezebrafishgeneap1γ2islocatedonchromosome2. The organization. domain core-ear boundaries of the zebrafish ortologue is similar to the human and the mouse genes, organized in 21 in also but orthologs respectively mouse and with it identity acids is 67% similar and amino to mammalian human 754 with identity of 66% shows protein protein predicted a The for (ap1γ2). encodes which sequence a zebrafish in revealed algorithm the of development of a multicellular organism. function the explore to model animal as zebrafish used We ubiquitously whilethesecondoneisexpressedinparticularnervoustissues. e.f imdcl cecs n Bohcooy nvriy f rsi, Italy; Brescia, of University Biothecnology and Sciences Biomedical Dep.of 91 1 , D.Zizioli γ-2 adaptin in the development of nervous system. Future rescue experiments will ap1g2) approach to better understand the γ1 and 1 , R.Bresciani γ2-adaptin subunits share high identity and the first one is expressed 3 etr o Bohmsr, er Ags Uieiiy Gottingen, Univerisity, August Georg Biochemestry, for Center γ2 subunit during development and its role in the machinery 2 subunit plays an important role during development of development during role important an plays subunit γ2 1 , E.Monti A search in ESEMBLE Genome Browser using TblastN Early developmentandgastrulation 1 γ, ß, μ and , A. Preti γ2-adaptin not only in the primary structure ap1γ2 transcript is detected in the embryos 1 -adaptin subunits (adaptor complex (adaptor subunits γ-adaptin , F. Cotelli γ2 function during development and σ. In human and mouse 2 2 adaptin subunit during subunit adaptin γ2 , P. Schu 3 2 prox1a prox1a e.f Biology, Dep.of γ2-adaptin γ1-adaptin probe as probe labelling codn t te xaso o te ok Ohr eeomna poess sc a ti bud tail as such processes, developmental Other yolk. the prolonged is of formation andsomitogenesis,occurwithoutmajordependence onyolksize. closure expansion ring germ the yolks, to large with according eggs in that such sequence developmental From these data we concluded that, in the evolution of teleost fish, variable yolk size has altered and tailofnormalshapesize. trunk head, the forming 24h, at normally develop They closure. ring germ before formed were somites diameter. 3-4 in early 15% the to embryos, 10 such by In expanded was yolk zebrafish injected The stage. blastula the at white egg chicken of injection by expanded artificially were Secondly, yolks size. egg normal zebrafish of eggs trout 5 than earlier is the which stage, aroundsomite 8 to at closure ring germ completed eggs such that observed and eggs trout rainbow of batches from eggs diameter) mm (~2.5 small exceptionally selected Firstly,we experiments. initiation of somitogenesis before germ ring closure depends on the yolk size, we conducted two marine tomato clownfish tropical the species, fish another in observed been also has embryogenesis of pattern Asimilar stage. somite 20 the around at closes finally ring germ the and embryos trout rainbow in stage) somite 10-20 (around stage epiboly late the at co-exist all somites and notochord bud, tail ring, Our species. fish such in sequential not are somitogenesis of initiation and formation bud tail closure, ring germ movement, epiboly Therefore, yolk. the of half covers only epiboly when starts somitogenesis and formed is structure bud-like tail a embryos, trout rainbow In yolk. the cover to epiboly for time longer a with associated be to seems which (<5mm) large is yolk egg the of size the trout, rainbow as such species, However,fish stage). other (somitogenesis in starts somitogenesis and for time short relatively epiboly a to takes it cover size, the yolk egg yolk small a have and which medaka, for and germ zebrafish In ring closure. Subsequently the tail bud is formed (bud stage) Plymouth, UK cmaio o htrcrnc eeomn o gr rn coue ti bd omto and T. formation Kudoh bud tail closure, ring germ somitogenesis betweenfishspecieswithlargeorsmalleggs of development heterochronic of comparison A Early developmentandgastrulation 1 School of Biosciences, University of Exeter,UK; of University Biosciences, of School 1 , E.Finch staining study using the mesoderm marker mesoderm the using study staining situ in 1 , C.Cruz Amphiprion frenatus which also has a large egg size. To examine if the 1 , J.Ghosh 1 , K. A. Sloman 2 School of Biological Sciences, University of University Sciences, Biological of School 2 , R.W. Wilson revealed that the germ the that revealed rt-ntl 92 1 163

Posters Posters preliminary resultsofChIP onchipexperimentsusingthisantibody. direct Toindentify targets of factors. Sox6, we have developed regulatory an antibody against the myogenic zebrafish protein. these We will report the on dependent of is activation progenitors that fast-twitch reveals Myf5 analysis. and expression MyoD bytransient of knock-down enhancer morpholino amuscle Simultaneous be to identified was element conserved specifically in the fast twitch fibres in the trunk and head muscles. Within this fragment a 220bp analysis, we have identified a identified have we analysis, that found have we antibody Prdm1a binds specifically anti-Prdm1 to sequences near to the with (ChIP) immunoprecipitation chromatin Using in theadaxialcellsofprdm1amutants. Consistent with this, as such genes, expressed in these fast muscle progenitors and to be capable of repressing slow muscle-specific be to found been has mouse, the in specification type fibre in implicated previously sox6gene, paraxial mesoderm does not express Prdm1a and differentiates into Prdm1a fast the twitch muscle of fibres. activity The the by protein, them expression of which is on induced by Hedgehog signaling. The imparted remainder of the myogenic character a type, fibre twitch slow the of are and cells adaxial the from derive embryo zebrafish the in differentiate to fibres muscle first The ofBiomedical Science,Department University ofSheffield, UK 164 X.G. Wang Regulation andfunctionofSox6inzebrafish skeletal myogenesis 1 nvriy f Singapore; of University nttt o Mlclr n Cl Booy Singapore; Biology, Cell and Molecular of Institute 93 1 , J.F. Chai Prox1, sox6 is not normally expressed in slow-twitch myoblasts but is de-repressed 1 , S.Elworthy smyhc1and 3 R Cnr fr eeomna ad imdcl eeis and Genetics, Biomedical and Developmental for Centre MRC eoi famn cpbe f rvn EF expression EGFP driving of capable fragment genomic sox6 3 , implying it to be a key regulator of fibre type identity. type fibre of regulator key a be to it implying stnnc, , W.O. Cheong 1 sox6 transcription start site. Using transgenic , andP.W. Ingham 2 et f ilgcl cecs National Sciences, Biological of Dept 1,2 Signaling in sox6 PRDM5 Regulates Wnt Signalling at Early Stages of Zebrafish Development. PloS ONE. 4(1); ONE. PloS Development. Zebrafish e4273. of Stages Early at Signalling Wnt Regulates PRDM5 N.,Meani PezzimentiF., G., M.Deflorian Mione and Alcalay M. (2009). The Suppressor Tumor Cancer. 6; 493-505. Grisendi S., Mecucci C., Falini B. and Pandolfi P.P. (2006). Nucleophosmintumor suppressoractivitiesthroughnegativemodulationofWnt signalling. and Cancer. their exert Nat. well Rev. as may and zebrafish in embryogenesis early in during signalling co-operate Wnt Npm1 repressing and Prdm5 that suggest results our So, functions. of convergence simple to due h peoye of phenotype the Wnt/β-catenin pathway in addition to the PCP pathway. canonical of repression in results that way a in development, of stages early at expression dkk1 both that showed approaches function of gain and loss using studies this, the overexpression of both of overexpression the this, with agreement In region. cephalic the of reduction a by pathway,accompanied Wnt PCP of block a to due gastrulation during movements morphogenetic of impairment an resemble and nooy onain Mln Italy; Milan, Foundation, Oncology nooy Erpa Isiue f nooy Mln Italy; Milan, Oncology, of Institute European Oncology, 1 G. Deflorian of wntsignallingduringzebrafish embryogenesis The tumor suppressor genes suppressor Thetumor Signaling bqios W fud ht h efcs of effects the that found We ubiquitous. the of members various which in domains to suppressor tumor somitogenesis as mid From embryogenesis. and during expressed strongly proto-oncogen as both acts 2006) that al., et and (Grisendi cytoplasm the and nucleus the between shuttles that protein nucleolar factor from transcripion a Prdm5, suppressor tumor Wnt/β--catenin and planar cell polarity (PCP) the that shown have belonging to the family of we PRDM proteins, is able work, to modulate, in a negative previous way, both canonical In and cancer, arefrequentlyassociatedwithchangesinWnt/β--cateninpathwayactivity. diseses by degenerative as such function, cell modulated stem altered from arise be may that states pathological to appear signals Wnt processes specific sets of these regulators that act at different levels of the signalling cascade. Moreover, all several In processes. developmental of vertebrate the members of of the expression of patterns temporal and spatial The embryogenesis. during roles Proteins encoded by the Wnt family of genes act as signals and have been shown to play essential Universita` degliStudidiMilano, Italy Cogentech, Consortium for Genomic Technologies,Genomic Italy;Milan,for Cogentech,Consortium assays, indicated that human PRDM5 associates with Nucleophosmin1 (NPM1), a (NPM1), Nucleophosmin1 with associates PRDM5 human that indicated assays, in-vitro knock down, suggesting an epistatic relation between the two genes rather than rather genes two the between relation epistatic an suggesting down, knock npn1 1,3 , N.Meani Wnt opat, whereas morphants, prdm5 family, their receptors and effectors suggest roles for these genes in a variety 2 , F. Pezzimenti and prdm5 and prdm5 4 iatmno i eiia Ciuga d Odontoiatria, ed Chirurgia Medicina, di Dipartimento 1,3 cooperate in the regulation the in cooperate (NPM1) nucleophosmin1 and prdm5 , M. Alcalay causes a “dorsalized” phenotype. Functional phenotype. “dorsalized” a causes npm1 Wnt pathways (Meani et al., 2009). Data obtained Wnt vrxrsin osn h phenotype the worsen overexpression prdm5 remains prdm5 whereas present, are family 1,2,4 agtd nc-on r similar are knock-down targeted npm1 npm1 synthetic mRNA is able to rescue , M.Mione 3 FMFR, nttt fr Molecular for Institute IFOM-FIRC, In zebrafish both genes are genes both zebrafish In . 2 Department on Experimental Experimental on Department expression localizes expression npm1 3 and prdm5 94 npm1 regulate 165

Posters Posters 1 K. Kawakami Organismic Organismic Biology, Academia Sinica, Taipei, Taiwan; 166 Han HW. diverse the tissuesincludingtheliver andintestine recapitulates line Tg(Nogo-B:GFP) zebrafish Transgenic iceia Sine, ainl awn nvriy Tie, Taiwan; Taipei, University, Taiwan National Sciences, Biochemical zebrafish Tg(Nogo-B:GFP) line recapitulates the endogenous expression pattern ofNogo-B pattern expression theendogenous mRNA. ThislinewillbeusefulforthestudyofNogo-Bgeneregulationduringdevelopment. recapitulates line Tg(Nogo-B:GFP) zebrafish the using Tol2-mediatedTransgenicthrough transgenesis. expression GFP generated drive to region was promoter same line Tg(Nogo-B:GFP) zebrafish transgenic A embryos. zebrafish and cells cultured both in functional be to demonstrated was it and clone BAC a from isolated was gene Nogo-B of region promoter 4.8-kb The intestine. and liver muscle, eyes, arches, brachial brain, the in expressed predominantly was mRNA Nogo-B that demonstrate to performed was hybridization situ in Whole-mount respectively. Rtn4-m/Nogo-gamma, and Rtn4-n/Nogo-beta alternative and usage promoter alternative RNARtn4-l/Nogo-alpha, as such ligands, Nogo other to identical are sequences These splicing. by generated are which Nogo-C1, and Nogo-C2, Nogo-B, termed transcripts, Nogo-related zebrafish three cloned we counterparts, mammalian In mammals, the Nogo family consists of Nogo-A, Nogo-B, and Nogo-C. Using sequences from Biology, NationalInstituteofGenetics, Mishima, Shizuoka, Japan nttt o Booia Ceity Aaei Snc, api Taiwan; Taipei, Sinica, Academia Chemistry, Biological of Institute 95 1,2 Y. Chen YC. , 4 , andCJ.Huang 1 B. Wu BK. , 1,2,3 1 C. Chu CY. , 1,2 C. Cheng CH. , 4 Division of Molecular and Developmental 1 G. Chen GD. , xrsin atr in pattern expression Nogo-B 3 nttt o Clua and Cellular of Institute 2 rdae nttt of Institute Graduate 1 S. . Hwang L. SP. , Signaling 3 , the significanceandroleofzGRbetainvariousphysiologicalconditions. GR of subset a on explore to order in undertaken is zGRbeta, overexpressing line fish transgenic a of construction glucocorticoids to response the of regulation target genes. the Further genetical manipulation, like in the use of isoform splice-blocking morpholinos and the this for role mRNA functional its a of shown microinjection has as embryos well zebrafish as in lines cell in vivo. in and vitro in both GRbeta zebrafish the of function zGRbeta of overexpression Transient found that the zebrafish GR (zGR) gene, can give rise to a GRbeta isoform. We have explored the therefore an animal model for studies on this receptor isoform was lacking. Surprisingly, we have and humans in demonstrated been only had recently,these GRbeta Until of diseases. occurrence the from suffering patients in resistance glucocorticoid and arthritis rheumatoid and asthma ligand-activated a Elevated as GRbeta levels have been acts associated with the occurrence which of immue-related diseases like alpha-isoform, GR transcription canonical factor, and the the GR beta-isoform, exist: which has a dominant-negative variants effect on GRalpha. splice two anti-inflammatory drugs. They exert their effects via the glucocorticoid receptor (GR). In humans, body’sthe as prescribed widely are hormones these of analogues Synthetic stress. to adaptation regulate to glands adrenal the from released are which hormones, steroid are Glucocorticoids Institute ofBiology, Leiden University, The Netherlands M.J.M. Schaaf, A. Chatzopoulou,P. Schoonheim,H.P. Spaink, The functionalroleofthezebrafish glucocorticoidreceptorbeta-isoform Signaling 96 167

Posters Posters L. Persani 168 P. Notch signalingregulatesthyroid development inzebrafish 1 Endocrinology, IRCCS Istituto Auxologico Italiano, Milan, Italy; early development, we are currently testing the co-expression of thyroid markers with Notch with markers thyroid ligands atvariousstagesofzebrafishdevelopment. of co-expression the testing currently are we the development, early treatments, thyroid in role direct these a plays signaling Notch if by define to order In signaling. Notch affected of activities seriously is structure vascularcell-autonomous non or cell-autonomous from either result could phenotypes thyroid observed pharyngeal the that Given UAS:Notch1a- x hsp70:gal4 of lines: intracellular domain(NICD),resultedinastrongreductionof thyroidfollicularcells. excess transgenic The two phenotype. intercrossing opposite by obtained the signaling, the to Notch in leads expressed function genes of of gain transcription Notch Conversely, increased thyroid. an showed signaling, Notch block to able inhibitor secretase gamma DAPT, a with treated morphology. Coherently, embryos modified a activate Notch signaling, we observed an expansion in the number of thyroid cells together with (pax2a, genes marker thyroid nkx2.1a, of expression the analyzed and signaling Notch over-activatedor blocked we formation, gland thyroid in role a plays signaling Notch whether Todetermine specification invivo. cells follicular in signaling Notch cell-to-cell development, direct of role gland the analyze thyroid to aim we affecting study this genes in identify to model animal excellent zebrafish an As represents disease. developmental multi-organ human a syndrome, Alagille in instance, observed, for is This development. vertebrate during organs and tissues different of formation the processes of cell fate decision. Mutations of Notch signaling members have been shown to affect and differentiation is represented by Notch signaling, a juxtacrine mechanism involved in several follicular cells specification are still largely unknown. A candidate pathway for thyroid formation early the underlying mechanisms the but identified, been have differentiation and development endoderm derived precursors cells. The main transcription factors and genes involved in thyroid of composed mainly is gland thyroid vertebrate The origin. genetic relevant a of support hypothesis the findings several if even undetermined, largely is pathogenesis CH years. recent the in newborns 1:2400 reach to reported been has (CH) hypothyroidism congenital of incidence The of Padua, Italy. Department of Medical Sciences, University of Milan, Italy; *Laboratory of Experimental Experimental of *Laboratory Italy; Milan, of University Sciences, Medical of Department Porazzi 97 and tg 1* 1* N Tiso N. , ) by mRNA by slc5a5) 2 D Calebiro D. , hybridization. In hybridization. situ in 1* F Benato F. , 2 T d Filippis de T. , mutant embryos, which fail to fail which embryos, mutant mib 2 Department Department of Biology, University * C Fritegotto C. , 2 F Argenton F. , Signaling 2 , 2 S. Jia and BMPsignalling balancing TGF-beta and augmenting by patterning dorsoventral zebrafish for required is Sipx Signaling 1 probably bydirectlyaugmentingandbalancingTGF-betaBMP signalingpathways. patterning dorsoventral embryonic in roles important plays Sipx Thus, dorsalized. are embryos likely that over-expression of co-injection of the embryos.Thedorsalizedphenotypesinducedbysipxover-expression couldbeinhibitedby over-expressionof hand, other the On embryogenesis. of role essential an suggests ventralization embryonic induced sipx is required for development of ventral tissues. The fact that of Knockdown embryos. sipx in vivo, we used mRNA over-expression TGF- and morpholino knockdown strategies in zebrafish the both of Tofunctions development. the embryonic understand of better stages early the enhances at distributed cells Hep3B Zebrafish mammalian expression. reporter Bmp-induced in and beta-induced Sipx of Over-expression bound Smad1/5. Sipx. also but is to Smad2/3, proteins with interacted binding only not Smad2/3 Sipx the that proved of experiments One Biochemical Smad2/3. with zebrafish interact several identified that we proteins screening, maternal hybrid two yeast Through factors. interacting their In view of important roles of Smad2/3 in early embryonic development, it is valuable to identify eatet f ilgcl cecs n Boehooy Tigu Uiest, ejn, China; Beijing, University, Tsinghua Biotechnology, ofMolecular andCellularDepartment Biology, and Baylor College ofMedicine, USA Sciences Biological of Department 1 , F. Dai 2 , D.Wu dnsmad3b mRNA encoding a dominant negative form of zebrafish Smad3b. It is 1 , X.Feng by antisense morpholinos dorsalized embryos, which implies that implies which embryos, dorsalized morpholinos antisense by sipx sipx induces ectopic TGF-beta/Nodal signaling, and as a result, the 2 , A. Meng 1 mRNA also caused dorsalization of dorsalization caused mRNAalso sipx sipx knockdown inhibited is maternally and globally and maternally is sipx for Bmp signaling during signaling Bmp for sipx 98 bmp2b- 169

Posters Posters 170 fulfill classicalsignaltransductionroles,butarenotabletointeractwithone-anotheranymore. to able variants STAT3.with and proteins BR1a endogenous between the replace will and we Next, phosphorylation Y705 transduction STAT3role. For of STAT3linear this of its functionality from integration purpose, signal the we untangle model to trying have are we Currently mapped the the aminoacids responsible that for the pathways. demonstrate interaction signaling consequent nucleartranslocationisoversimplified. two least data at of integrator our signal a Furthermore, as acts – complex found newly the of observed These data suggest that usually STAT3 – potentially together the with other thus far unidentified components occurs, observed. Y705 of was exclusion nuclear instead but phosphorylation found, STAT3not of was accumulation nuclear by consequent STAT3 BR1a.While of with activation theinteraction theoretical way through anon-common in influenced STAT3 is factor transcription latent the of localisation subcellular Interestingly,the administration. ligand BMP the activation of STAT3, regulated via phosphorylation at Y705, is actively altered in response to Co-immunoprecipitation assays showed that STAT3 physically interacts with BR1a. Furthermore, this Within sources. far unconnectedsignalingpathways. studied all from isolated be can complex, we always found BMP-receptor 1a (BR1a) and that STAT3 complex – two proteins belonging to thus protein identify (>1MDa) to able large were a we counterparts, embryonic respective their cells and stem mouse cultured the of medaka, activities of signaling comparing at aimed experiments of series a During BMP is There a concertedresponse. fashion. isolated produce to integrated linear,are antennae of array massive a from rather inputs where hub, communication a in displayed a to similar reality,more In likely on. is so cell and a signaling MAPK signaling, SHH signaling, often are cascades Signaling Biozentrum, Physiological I, Chemistry University of Wuerzburg, Germany T. U.Wagner, S.Meierjohann,M.Kraeussling,E.Thoma,Schartl Characterisation ofafunctionalphysical interaction betweenSTAT3 andBMP-receptor1a 99 Signaling 237(1):222-232. are duplicated and dynamically expressed during zebrafish embryonic development. Dev. Dyn. M., Milanetto 1) and theywerebredtoobtainEmilin3nullmicewhicharecurrentlyunderphenotypicanalysis. clones, independent two from obtained were mice chimeric Several females. foster in implant were used for generation of chimeric mice by microinjection of ES cells into host blastocysts and Six occurred. had them of four characterization karyotypic additional after and identified, were clones ES positive event recombination homologous correct a which in clones identifying for clones were isolated; 995 of these individual cell clones were further expanded and investigated a with cells ES R1 transfected We experiment. large-scale a of means by performed approach, targeting gene Emilin3 new a out carried we Therefore, clones. targetedcell of ES instability karyotypic to due probably reached, not was generation F1 the to allele inactivated the of transmission and low extremely is gene this of recombination homologous of frequency The difficult. particularly is in undertaken gene Emilin3 the alreadyof inactivation targeted emilins, other unlike was that, revealed attempts previous cells (ES) of stem generation However, embryonic laboratory. murine our in inactivation gene The Emilin3 targeted oocytes. years, past the fertilized In emilin-3. for in coding gene the on oligonucleotides concentrated were mice morfolino in studies or mRNA of microinjection by studies regulate emilin-3 expression during early development. Moreover, we are performing functional they whether check to pathways relevant other investigating Wecurrently (swirl). are BMP2 for or (dino) Chordin for mutant embryos in normal appears genes Emilin-3 two the of expression Additionally,treatments. drug by blocked were pathways molecular these which in embryos of does not appear to be expression regulated Emilin-3 by (1). either development Hedgehog, early FGF during or roles Notch key play signaling, may as emilin-3 indicated that by suggesting ISH genes coding for emilin-3 are abundantly expressed at early stages in notochord and floor plate, other emilin genes reveal a mesenchymal or cardiovascular expression profile, the two zebrafish While domain. gC1q C-terminal the lacking family the of component only the emilin-3, of one the is pattern distinctive most The (1). processes morphogenetic certain in role a play that may they suggests notochord, posterior the or system nervous central the as such areas, peculiar in and site-specific patternsoverlapping mostly including mesenchymalwith structures. Expression ofdevelopment, some emilinembryonic genes during manner spatio-temporal a in regulated Whole-mount has gene ortholog. human the with syntheny and structure conserved emilin showing zebrafish, in paralogs two Each zebrafish. in family this of members eight identified we tools, similarity fish the in emilins of expression the investigated we vertebrates, structural common with and other emilins matrixwere recently isolated in mammals. To extracellular gain insight into prototype these fibers, proteins elastic with the in association The in lower organs murine and human in family,found domain. this is of Emilin-1, of cysteine-rich N-terminal glycoproteins characteristic a containing of and organization family a are Emilins Dept. of M. Milanetto Characterization ofexpressionandfunctionnovel emilingenesinmouseandzebrafish Signaling 1 Histology, andMedicalBiotechnologies, Microbiology 1 , N.Tiso hybridization (ISH) revealed that expression of zebrafish emilin genes is genes emilin zebrafish of expression that revealed (ISH) hybridization situ in targeting construct and, after double positive-negative selection, 1505 selection, positive-negative double after and, construct targeting Emilin3 Tiso N., Braghetta P., Volpin D., Argenton F., Bonaldo P. (2008) Emilin genes Emilin P.F.,P.,(2008) Bonaldo Argenton VolpinBraghetta D., N., Tiso 2 , A. Schiavinato 1 , P. Braghetta ncot ie a uscesu ad these and unsuccessful was mice knockout Emilin3 1 , D.Volpin 2 1 Biology Univ. ofPadova, Italy , F. Argenton . Using sequence Using . rerio Danio 100 2 andP. Bonaldo 171 1

Posters Posters this hypothesis in accordance with the role of cat1 in the development of the gills in the cat1 the in zebrafish morphants. gills the of development the in cat1 of role the with accordance in hypothesis this stall or due to lack of collagen pharyngeal arches might not develop. We will further investigate Based synthesis. collagen the on this we hypothesize that for if there is lack of arginineimportant then the branchial cell proliferation might is l-proline and proliferation cell in important polyamines are The l-proline. and polyamines into metabolized be further to can L-Arginine which converts L-ornithine Arginase synthase. oxide Nitrous to availability its through production NO determines it because mainly pathways, signaling cellular of homeostasis regulation L-Argininethe in and involved is transport, to L-Arginine in head involved shortened is by Cat1 of characterized distance. arches trunk was pharyngeal thoracic which the properly dpf developed 5 not and dpf were 3 morphants At the absorption. yolk reduced showed and embryos axis morphant body the bent hpf a 24 At gill knock-down. morpholino in blocking appearing splice by started done expression the (dpf) fertilization post arches. The expression levels days in gills increased at 5 dpf. Functional characterization of cat1 was 3 At stronger but muscles. widespread somite was expression in the hpf 24 At hybridization. in-situ mount whole by cat1 expression was first detected in somite muscles and eyes around 16 h post fertilization (hpf) The cat1. zebrafish of role developmental and expression early the investigate to was study this mammals, In retro-virus. ecotropic for CAT-2, receptor of proteins, aim related CAT-2A, aThe additional CAT-3,identified. three and been as have acts also and proliferation for essential survival, be to cell shown been has and expressed ubiquitously is It arginine. and lysine acids 1 172 S. Narawane 1 Transporter acid Amino Cationic of (CAT1) inzebrafish (Daniorerio) role developmental and distribution Spatiotemporal A1 s Na is CAT1 the of member a is CAT1 University ofBergen, Norway eatet f oeua Booy Uiest o Bre, Norway; Bergen, of University Biology, Molecular of Department 101 + 1 needn tasotr f ainc mn ais icuig h esnil amino essential the including acids, amino cationic of transporter independent , S.Ellingsen 1 solute , A. Fjose arrier 7 family (SLC) of proteins, and is also called SLC7A1. called also is and proteins, of (SLC) family 7 carrier 1 , I.Rønnestad 2 2 eatet f Biology, of Department Signaling differentiate. to beta-cells permit to required be of would amount dose lower while high differentiation a alpha-cell allow fate: cell pancreatic for critical is embryo the present proteins in nkx6 of dose global the that is investigating currently are we hypothesis One as inaalmostcompletedisappearanceofglucagonexpression. the double knock-down of the two genes results in a clear decrease of expressing cells, while the expression of their cells, same the in expressed First pancreas. the to segregate, rapidly domains restricted expression be to before endoderm The knock-down of knock-down The cells untilatleast24hpf. never expressed in the mature hormone-producing endocrine cells, zebrafish, In in normally develops pancreas the pancreas and deficiency for Nkx6.1 results in a specific abrogation of beta-cell neogenesis while The homeodomain proteins Nkx6.1 and Nkx6.2 have been shown to be expressed in the murine GIGA-R, Ulg, Liège, Belgium A. C.Binot,M.Winandy, IsabelleManfroid,P. Motte,B.Peers,M.L.Voz nkx6 genescontrolendocrinepancreaticcellfateinzebrafish Signaling double-mutant embryosdisplayaseverereduction and nkx6.1 or of or nkx6.1 r epesd t h bgnig f h smtgnss n the in somitogenesis the of beginning the at expressed are nkx6.2 leads to a striking decrease of the number of number the of decrease striking a to leads nkx6.2 single-mutant Nkx6.2 being expressed more ventrally.While more expressed being nkx6.1 insulin, somatostatin or

in alpha-cellnumber.

ie I contrast, In mice. ghrelin is not affected. In contrast, would be necessary to necessary be would nkx6 102 nkx6.2 is expressed in beta- insulin expression as well Nkx6.1/Nkx6.2 glucagon­ is nkx6.1 173 -

Posters Posters 174 of NCmutantsandtheirwild-typesiblings. Once combined, we expect these systems will allow us to compare NCC sox10:cre transgeniclines. development in cultures a in expression Cre gives system a of tests Initial culture. in NCCs of labelling permanent lost as NCCsdifferentiate.Consequently, rapidly wearemodifyingthesystemtouseCre/Loxfor is expression GFP since difficult is progeny their and cells in individual tracking lines that using the are of control we the approach, under this driven of is GFP tests which initial For genes. mutant of effects autonomous us cell- the testing allow for useful be will will particular,it in embryo; approach single a from this derived NCCs study anticipate to We embryos. non-transgenic disaggregated, of pool a a problem in this cultured was solve NC tranhsgenically-labelled to with embryo order single a In where devised differentiation. was strategy significant and for survival need NCC the for highlightingnumbers cell thrive, cultured to failed also embryos single disaggregated from However, embryos. zebrafish disaggregated cultures Likewise, medium. the in factors growth of lack to due probably attachment/growth, in of pool a from failure a in resulted density clonal at NCCs to or NCCs single to conditions these of application differentiation derivatives and many survival and achieved NCCs we of conditions, culture these Using conditions. culture NCC mouse and avian with began we cultures, cell NC zebrafish published of absence the In NCC mutantsincellculturewouldopenupnewpossibilitiesfortestingofgenefunction. to system model study a developmental processes such as specification and differentiation.used The study of individual is and system nervous peripheral the of glia and neurons and cells complement to in vivo studies. The NC is responsible study for the several for cellular allow lineages that such as pigment in vitro has been less keenly pursued. The purpose of this project is to develop culture conditions Whilst in vivo analysis of zebrafish mutants is widespread, the ability to study these phenotypes Biology andBiochemistry, University ofBath, England F. Rodrigues,D.Tosh, andR.N.Kelsh An invitroapproach forthestudy ofzebrafish ncmutants 103 of the phenotypes of zebrafish neural crest (NC) mutants, as a as mutants, (NC) crest neural zebrafish of phenotypes the of vitro in -like pattern. We are now attempting to generate stable generate to attempting now Weare pattern. sox10-like promoter. This system has the limitation the has system This promoter. sox10 construct indicate that this that indicate construct sox10:cre Signaling 1 The efficiency of rescue of Ptc fusion protein whose expression is driven by 13 kb upstream of Ptc1 translation initiation In site. an be effort to identify the can functionally important domains animals of the Ptc proteins, transgenic we have generated which with ease the generated toinvestigatevariousaspectsofthesignalingprocess. and zebrafish in mutants pathway Hh advantage the taking are Weof Fused). (eg mammals and Hip) (eg Drosophila both from absent are that components pathway of function the retain they as signaling, post- Hedgehog of analysis the for in model interesting an and present Zebrafish system. unusual and complex this development of understanding our in animal in gaps subsequently still are there pathway and signaling in Hh the of Drosophila components core the uncovered in role has mouse analysis mutant fundamental Although a homeostasis. tissue plays embryonic signaling (Hh) Hedgehog Sheffield, UnitedKingdom Funded by Wellcome Trust grants074974and082962 analysis ofGli2distributioniniguanamutantembryos. our of results the present Wewill embryos. the type wild in of nucleus the tip to as the well mutant as to cilia primary localises and Gli2 type that find wild we in cells, cultured factor in transcription reported previously key As embryos. this of distribution cellular sub the study to this used and Gli2 of form tagged GFP a encode to clone BAC genomic a inHh engineered have we protein theiguana/DZIP1 for role activity.protein Gli of control essential the in it Toimplicating signaling, further,function this investigate an identified have we studies, previous In domain isremovedsuggeststhatC-terminalofthePtc1proteinregulatesitsturnover. erbooy Kns olg Lno, ntd Kingdom; United London, College Kings Neurobiology, H. R.Kim Analysis oftheHedgehogsignalingpathway inzebrafish Signaling Biomedical Science, University of Sheffield, United Kingdom;Sheffield, United of Science,University Biomedical 1 , J.Richardson 2 , P.W. Ingham ptc1 mutants using full length Ptc1 and Ptc1 in which the C-terminus 3 3 imdcl cec, nvriy of University Science, Biomedical 2 MRC centre for Developmental for centre MRC 104 175

Posters Posters 176 and disorganization ofthemigratingpLLp. eventually leads to failure of FGF signaling and unregulated this Wnt and signaling. atoh1 express This cells leads many too to fails, collapse signaling Notch When signaling. FGF regulating in We neuromasts. forming the in have now atoh1 shown that the central atoh1 of expressing cell in the the neuromast expression also plays a both critical role and determines rosettes which epithelial domain, of trailing morphogenesis adjacent the in center signaling FGF a and edge leading the at center signaling Wnt a centers; signaling antagonistic mutually by coordinated is mediated-lateral Notch deposits by determined periodically cell inhibition. hair The generation of and new atoh1-positive neuromasts and their centraldeposition as the pLLp caudally a migrates caudally has the within pLLp, migrates formed neuromast, migrating Each tail. (pLLp) and trunk zebrafish the primordium in skin the under line neuromasts lateral posterior The LMG, NICHD, NIH, Bethesda, USA M. Matsudaand A. Chitnis the in migration and morphogenesis fate, lateral lineprimordium cell coordinating for essential is signaling Notch 105 Signaling Organogenesis, Centre de Regulaciò Genòmica, Barcelona, Spain;Barcelona, Genòmica, Regulaciò de Centre Organogenesis, Hbeh Isiue Urct te Netherlands; the Utrecht, Institute, *Hubrecht P. van Duijn*, A. Faucherre Unraveling ptenfunctioninzebrafish development Signaling development. fluorescently using embryos zebrafish labeled pten developing proteins primordial will be the used the to of increase our of understanding imaging of the development role live of the pten addition, in embryonic in In pten cells. of germ role specific the into insight more provide will mutant fish with the transgenic VASA:EGFP line, which specifically express EGFP in their PGCs, the of development the primordial affect germ cells to (PGCs). Detailed seemed analysis of also the offspring expression resulting from pten crosses of of our pten loss embryos mutant double not show embryonic phenotypes, double mutant embryos died at 5 days post fertilization. In the Whereas embryogenesis. during functions redundant have to In the zebrafish genome two PTEN genes, PTEN two genome zebrafish the In development. embryonic in PTEN of function the investigate further to zebrafish used have we processes developmental studying for model ideal an being embryogenesis. As during early die found to be crucially important for normal embryonic important development cell biological processes, such as as proliferation, survival and homozygous migration. PTEN was also on the PIP3 level and as a consequence they counteract each other’s actions in the regulation of of PIP3, a lipid second messenger produced by PI3K. PTEN and PI3K thus have opposing effects The tumor suppressor PTEN primarily acts as a lipid phosphatase by specifically dephosphorylating Leiden University, theNetherlands # , S.Choorapoikayil*andJ.denHertog* ptena and # aoaoy f esr Cl Booy & Biology Cell Sensory of Laboratory were identified, which were shown were which identified, were ptenb -/- or ptena-/- 106 $ $ Institute of Biology Leiden,Biology of Institute -/- mutants did mutants ptenb-/- Pten null mice 177

Posters Posters 178 are embryos zebrafish in (hpf)) fertilization Calcium post acetylcholine-driven by (Ca hours induced contractions 22 muscle spontaneous - by characterised (17 movements earliest The School ofBiologicalandChemicalSciences, QueenMary, University ofLondon, UK M. Lahne,C.KrivcevskaandR. Ashworth signalling contraction‑dependent mechanism a by regulated is muscle skeletal in organisation Myofibril 1. appear toberegulatedbyanovelcontraction‑dependentmechanism. movement, embryonic upon rely which differentiation, muscle of development. steps late fibre the conclusion, muscle In during organisation myofibril for required is apparatus contractile functional a that Takensuggest intact. data together,remained our coupling E‑C of component Ca is controlled are currentlystillunknown. mechanisms signalling organisation intracellular myofibril underlying The that depolarisation. propose membrane of we [1]. downstream mutants myofibrils these from misaligned evidence and lengthened, on Based sarcomeres shortened specifically development, muscle receptor,skeletal dihydropyridine disrupted or show receptor acetylcholine the namely muscular communication by measuring Ca apparent underlying cause of the observed myofibre phenotype. Nonetheless, assessing functional anneuro- but embryos, disruption of neuronal pathfinding blebbistatin‑treated might affect E‑C coupling upstream of contraction and be the and control in increase in neurite number and the loss of varicosities was observed during drug treatment. The similar was outgrowth Axon myosin non‑muscle and muscle E‑C both inhibits blebbistatin as lacking However, role. regulatory critical the mutants a plays contraction to similar myofibrils misaligned immunocytochemisty. Slow muscle fibres of blebbistatin‑treated embryos (10, µ50 M) displayed niio bebsai t bok uce otato. oeet a bokd o a eid of period a for blocked was Movement contraction. 7 muscle block to blebbistatin inhibitor myosin the using approach pharmacological a employed we mechanisms two these between distinguish depolarisation. To membrane of downstream organisation myofibril in involved be hours starting at 17 at starting hours 2+ 107 2+ mediated muscle contractions or Ca or contractions muscle ‑mediated ) signals. Mutant lines lacking key components of the excitation-contraction (E‑C) pathway, JCellSci,2005.118(Pt22):p.5181-90. in thezebrafish embryo. Brennan, C., et al., II, it might affect not only muscle fibre but also neuronal development. neuronal also but fibre muscle only not affect might it II, hpf and skeletal muscle morphology was subsequently examined using examined subsequently was morphology muscle skeletal and hpf Acetylcholine and calcium signalling regulates muscle fibre formation 2+ 2+ signals acting independently of contraction could contraction of independently acting signals signals in muscle fibres revealed that the excitatory opig sgetn ta muscle that suggesting coupling, Signaling II rvos tde hv ivle te rmr clu i te aac bten h Wt beta- Wnt- the between balance understand thefunctionofRpgrip1landmoregenerallycilia inthisprocess. the in cilium primary catenin andtheWnt-PCP pathways(reviewedin1).Thepresentstudyshouldallowusto better the involved have studies Previous morphants. in pathway this of components different of localization subcellular the and abundance the assaying currently are we hypothesis, this test location. To specific this at function their or the at described cilium been the to traffic their regulate to be could Rpgrip1l of function a cilium, primary have the of base pathway this of components several Because pathway. Wnt-PCP the rescue to trying and We are currently looking for a physical interaction between Rpgrip1l and different PCP proteins, genetic interaction of a demonstrate could pathway.we (PCP) Indeed, polarity cell Wnt-planar the in involved genes of gain-of-function and loss- in found often convergence-extension, in defect a of reminiscent an and tail human shorter the of a injection by rescued displayed be could embryos and dose-dependent was phenotype: phenotype The same notochord. undulated the to led morpholinos different two of Injection analysis. knock-down for morpholinos designed and orthologue zebrafish the detail greater in address to order In the anomaliesfoundinfœtuseswithMeckelsyndrome,mostsevereformofciliopathy(3). the of phenotype The syndrome. Meckel and B type syndrome the in mutations gene in the mouse has shown its involvement in Hedgehog cilia (Hh) signalling (2). We in recently found The alterations mouse, the In randomisation ofleft-rightasymmetry, anddefectsindorsoventralpatterningofthespinalcord. 1). in (reviewed roles physiology diverse formation or function lead to a range perform of developmental defects, such as and polydactyly, to renal cysts, evolved development have animal and cells in eukaryotic of surface the from project Cilia CNRS UMR7622, INSERMERLU969, Université etMarieCurie, Pierre Paris, France S. Schneider-Maunoury andC.Vesque Function oftheciliarygeneftm/rpgrip1lduringzebrafish development Signaling (3) (2) (1) Ftm/Rpgrip1l gene codes for a protein localised in the basal body of cilia. Inactivation of this Delous etal.,.2007.NatureGenet.39(7):875-81 Vierkotten etal.,2007.Development.134(14):2569-77 Gerdes etal.,2009.Cell137,32-45. N, hc i nt agtd y h mrhlns Ti peoye is phenotype This morpholinos. the by targeted not is which RNA, RPGRIP1L ee n w hmn iiptis ih ifrn svrte, Joubert severities, different with ciliopathies human two in gene RPGRIP1L rpgrip1L with the core PCP gene rpgip1l morphant phenotype by injection of active components of the of components active of injection by phenotype morphant Ftm/Rpgrip1L function in signalling pathways, we cloned we pathways, signalling in function vangl2 in double knock-down experiments. mouse mutant recapitulates mutant mouse Ftm 108 rpgrip1l 179

Posters Posters we suggestthattpbglisanimportantregulatorofWntreceptorcomplexformationoractivity. Wnt dependent beta-catenin the of pathway,inhibitor feedback a as role its to contrast in Intriguingly, beta- catenin signaling,counteractingwnt8functioninbrainpatterning. of inhibitor required a is tpbgl endogenous that show embryos zebrafish in experiments Knock-down complex. receptor Wnt the of level the at signaling beta-catenin with interfere to forcedby induced signaling inhibits tpbgl Thus, LRP6. and Frizzled of interaction Furthermore, beta-catenin. stabilized or Axin1, media, dominant-negative Wnt-conditioned Gsk3b, dominant-negative by by activation activation pathway with signal interfere cannot it with since complex, destruction the interferes of upstream and it since cells, receiving Wnt in overexpressed cells. mammalian cultured and embryos zebrafish in both ligands that Wnt by activated found we Interestingly,function. its about known is little carcinomas, of variety a in upregulated is and vertebrates in Although stages. developmental and tissues many in signaling beta-catenin by regulated is domain, extracellular glycosylated a with protein transmembrane single-pass a zebrafish embryogenesis. We found that found embryogenesis. We zebrafish development, zebrafish lines and microarray-based transcriptional profiling to identify Wnt animal target genes during during Wnt mediating Weknown. processes are function transgenic Dkk1, in of Wnt8 overexpression and inducible Axin1 used genes target few roles of its of many for plethora yet regeneration, and homeostasis a tissue regulates signaling Wnt/beta-catenin Netherlands 1 180 G. Weidinger Feedback Modulationof Wnt Signalingby Trophoblast Glycoprotein-like itcnlg Cne, rse Uiest o Tcnlg, Germany; Technology, of University Dresden Center, Biotechnology 109 enhances beta-catenin independent Wnt signaling in zebrafish embryos. Thus, embryos. zebrafish in signaling Wnt independent beta-catenin enhances tpbgl 1 , B.Kagermeier-Schenk 1 , andH.Spaink trophoblast glycoprotein-like ( trophoblast oety niis eactnn signaling beta-catenin inhibits potently tpbgl 2 , a gene encoding gene a tpbgl), 2 edn University, Leiden is conserved is tpbgl appears tpbgl Signaling acts Tpbgl deficits inM-cellfunction. the absence of In PKCγ, this M-cells. normal developmental zebrafish paradigm does on not occur synapses and fishexcitatory exhibit major into AMPAreceptors of insertion and recruitment developmental normal the to leads PKCγ of activation that suggest results Takenour together, deficits intouchresponsesandwereunabletohatchoutof the chorion. mature fish at 48 hpf or 72 hpf. Moreover, fish injected with PKCγ morpholinos exhibited severe developmentally the than rather hpf 33 at embryos young the like much characteristics mEPSC were incapable of trafficking AMPA receptors to the cell membrane, and which exhibited overall that fish in resulted PKCγ against morpholinos of Injection membrane. synaptic the to receptors which increases synaptic activity, suggesting that activating PKCγ leads to the trafficking of AMPA receptors. All of these agents were capable of blocking the effect of PMA and a high K high a PMAand of effect the blocking of capable were agents these of All receptors. B), (latrunculin AMPA with PICK1, polymerization and NSF proteins trafficking actin the of association of the inhibit that peptides blocker and selective a applied we amplitude, mEPSC in to block the effects of PMA. Toof determine form if traffickingactive mechanisms that werethe involved found inof the increase application and PKCγ PMAIntracellular of PKCγ. effects of the levels for high relatively responsible express be M-cells might that isoform PKC the identify to immunohistochemistry Weused action. of locus postsynaptic and presynaptic a both has PKC 1), blocked the effects of DOG and post- PMA on mEPSC amplitude but not frequency, hour indicating that 30 frequency (~2.5-fold). Pipette and from (~1.5-fold) application amplitude of peak the mean mEPSC specificincreased age PMA PKC and inhibitor,DOG Both PMA. bisindolylmaleimide excitatory and in IDOG (BIS miniature ranging AMPA M-cells record to fertilization from (hpf) to 72 hpf. AMPA mEPSCs were recorded in the presence of the PKC activators, electrophysiology (AMPAR-mEPSCs) clamp currents patch postsynaptic cell whole used We determine ifPKCmightbeinvolvedinthemodulationof AMPARs. protein that to sought we Therefore, indicated zebrafish. developing is of M-cells results in expressed highly (M-cells) is recent (PKC) C kinase neurons our Mauthner Furthermore, AMPARs. zebrafish by in predominantly transmission mediated synaptic excitatory fast that shown plasticity.synaptic and transmission synaptic Wein receptors previously these have of function CNS. vertebrate the in the receptors controlling for mechanism important an be of AMPARsto Phosphorylation shown been has excitatory major are (AMPARs) receptors AMPA Glutamate ofBiological Sciences,Department University of Alberta, Edmonton, Canada DW. Ali andS.A.Patten Mauthner zebrafish on neurons synapses glutamate of development normal the for required is PKCγ Signaling V5-3) was able was γV5-3) ( peptide inhibitory PKCγ the and amplitude mEPSC in increase an to led 110 + saline, 181

Posters Posters opat o te o-aoia Wt inln ptwy sc as such pathway, signaling Wnt non-canonical the of morphants non-canonical the between the interactions synergistic through pathway.found WeWnt signalling by CE regulates Antr2a gastrulation. during movements (CE) extension and convergent mediates Antr2a that indicate knockdown gene (MO)-mediated CMG2. of human functions the the of of homolog Analysis closest the Antr2a being Receptors, Toxin Anthrax encoding genes 6 contains Zebrafish vertebrates. in found only is CMG2 that revealed normal analysis Genome its However, interactions. cell-cell mediate physiological and/ordevelopmentalfunctionsarestillunclear. potentially also matrix could extracellular bind and to proposed components, been has that domain A type Factor Willebrand von to related highly extracellular an with protein membrane I type a is CMG2 syndromes. recessive autosomal two (ISH), hyalinosis systemic infantile and (JHF) fibromatosis hyaline juvenile with in an bind that receptors surface cell directly to two Anthrax toxin and supports cellular intoxication. the CMG2 was found to be upregulated of one is 2) Protein Morphogenesis (Capillary CMG2 Germany 1 Fédérale de Lausanne, Switzerland, 182 I. Castanon extension duringgastrulation convergent wnt-dependent mediate to LRP6 with interacts 2A/CMG2 receptor toxin Anthrax to mediateCEmovementsduringgastrulationpresumablyby regulatingcell-cellinteraction. accumulates intracellularly. Taken together these results indicate that Antr2a interacts with LRP6 in while sites, contact cell-cell at Antr2a of increase an Wnt11showed overexpressing cells embryonic uniform, possible role in mostly regulating cell-cell interaction. Moreover, this polarization is is Wnt11-dependent: membrane plasma the Antr2a appears at clearly polarized. Antr2a localization is enriched at the sites LRP6 of cell-cell Rab11-positivecontact, suggesting while and a However, Rab5-, intracellular endosomes. in as well as cells embryonic zebrafish plasma of the membrane to localize Antr2a and LRP6 Both morphants. double antr2a/lrp6 the in enhanced observation, this with receptor-related Consistent (Low-density-lipoprotein zebrafish. LRP6 in receptor 6) protein Pull-down experiments indicate that Antr2a binds to the has beenshowntoregulatethecytoskeletalarchitecture duringvertebrategastrulation. typical canonical-Wnt signalling pathway injection of of injection the from resulting phenotype CE Conversely,the Biochemestry,Geneva,Universityof Switzerland, 111 in vitro model for human capillary tube formation, and mutations in this gene are associated ), Antr2a is mostly absent from the plasma membrane and membrane plasma the from absent mostly is Antr2a (slb), embryos mutant wnt11 1 rhoA mRNA, a downstream component of the non-canonical Wnt signalling, which , L. Abrami opat as soe a n1-eedn C peoye wih is which phenotype, CE Wnt11-dependent a showed also morphants lrp6 2 , CP. Heisenberg uig erfs ebync eeomn bymorpholino development embryonic zebrafish during antr2a 3 Max-Planck Institute of Molecular Cell Biology an, Dresden, 3 , G.vanderGoot 2 Global Health Institute,Health Global PolytechniqueEcole 2 , M.Gonzalez-Gaitan morphants and mutants/ and morphants anthr2a MO could be rescued by rescued be could MO antr2a wnt11, and wnt5a 1 Signaling knypek. of Lrmpprotein. source localized highly a providing by possibly development, early during organelles these of to centrosomes and/or the nuclear periphery may be critical for proper function and distribution spread diffusely throughout the blastodisc. Our results suggest that recruitment of distinct localization pattern is completely lost in with nuclear membranes and centrosomes, and is later found on the spindle during mitosis. This centrosomes as early as 15 mpf, prior to pronuclear fusion. During prophase, that showed staining antibody gamma-tubulin and hybridization variant found in zebrafish appears to be significantly larger than variants described inadult described thanvariants larger of besignificantly Analysis species. other of to tissues appears zebrafish of in found function variant molecular the about known is little Very mutation was detected in the longest transcript of cloning Positional fertilization. four that contains two genes. Both genes were sequenced in their entirely and a single missense after shortly lost be the of location the down narrow to us allowed to appears centrosome and pronucleus male of Additionally, apposition fertilization. close after the shortly morphology and dynamics that show normally during early cell divisions even though DNA is absent from most Interestingly, chromosomes. segregate proceeds cytokinesis embryos, mutant of majority the dividing in cells. Here we properly to fail consequently and spindles, mitotic form never fusion, and migration pronuclear stages in zebrafish. Embryos from homozygous mutant developmental early during acting products maternally-supplied for screen gynogenesis-based mutation maternal-effect recessive The formed zygoticnucleusinpreparationforthefirstmitoticdivision. migration. After pronuclear fusion, duplicated centrosomes align on opposite sides of the newly between the sperm pronucleus and the nascent centrosome is maintained throughout pronuclear pronucleus, which then begins a rapid migration components, toward the centrosomal male pronucleus. maternally-derived Amonoaster.sperm a nucleate female the with contact makes eventually monoaster growing The close to with association begins along envelope centrioles, pronuclear the sperm-derived and the expands nucleus form, sperm the As centrioles. of pair a it During normal fertilization, a compacted sperm nucleus enters the egg cytoplasm bringing with ofGenetics,Laboratory University of Wisconsin, Madison, USA RE. LindemanandF. Pelegri jaw1 withessentialrolesinnuclear-centrosomal attachment andpronuclearcongression Positional cloning reveals that Signaling fue embryos develop a normal monoaster,normal a develop embryos membrane nuclear abnormal manifest but futile cycle transcript localization in wild-type embryos by embryos wild-type in localization transcript lrmp futile cycle ( futile encodes a maternal gene homologous to human mutation to a 56 kb region on linkage group linkage on region kb 56 a to mutation fue fue embryos, where the transcript appears to be lymphoid restricted membrane ( protein ) was previously identified as part of a of part as identified previously was fue) fue females (fue embryos) fail to undergo n n sse, n te maternal the and system, any in lrmp mRNA colocalizes with colocalizes mRNA lrmp 112 lrmp is associated lrmp transcript in situ in lrmp). lrmp/ 183

Posters Posters 184 thereby molecules, regulatory potent these promoting preciseandaccuratemigration. of patterns expression the refine and modulate to level transcript the at acting is repression miR-430-mediated which in model a support results Our meeting. the at presented be will results these and CXCR7, antagonist the of regulation the analyzing currently Weare line. germ the to migration cell germ affected SDF1a of protection the functional relevance of miR-430-mediated regulation of each individual target. Interestingly,specifically to shown been have and interfere withtheindividualmiRNA-target interactions.Thisapproachallowedustoinvestigate site target miRNA the to complementary are target protectors These morpholinos. TargetProtector used we migration, cell germ for regulation this of CXCR7b mRNAs are likely regulated by miR-430 in vivo. To assess the physiological significance reporters repression in wild type GFP-3’UTR embryos. Taken the together, these of results GFP-reporter indicate abolished that site targetSDF1a, CXCR7a, expressionmiR-430 and putative the of the mutation (iii) and genes enhances three all for function dicer of loss (ii) in 3’UTRs, sites target their miR-430 putative have CXCR7 and CXCR4 SDF1, (i) that revealed analysis Our CXCR7 is believed to SDF1. ligand be secreted the of a gradient a follow non-signaling and CXCR4 receptor receptor the express cells responsible Migrating for refining SDF1 protein expression. miRNAs. by regulated are pathway this of components germ any whether assayed we migration, guides cell signaling chemokine Because miRNAs. of absence the in affected was migration Analysis ofthecellmigrationphenotypeinMZdicermutantsrevealedthatprimordialgerm is miR-430, particular, in miRNA one that ubiquitously expressed shown in has early laboratory depleted development Our are and microRNAs. plays mutants important mature These roles of MZdicer). in dicer, embryo zygotic morphogenesis. and miRNA the (maternal in dicer deficient enzyme embryos processing used we development, zebrafish during miRNAs of role the Tostudy translation. and degradation mRNA regulating by expression gene shape MicroRNAs ofGenetics,Department Yale University, New Haven, USA A. A. Statonand A. J.Giraldez Regulation ofChemokineSignalingby themicroRNA miR-430 113 Signaling during vertebratedevelopment. activity Wnt/beta-catenin proper for glypicans for requirement specific unexplored an is there Notum3 functions as part of an autoregulatory loop by which Wnt signaling is restricted and that Wnt/PCP signaling or the activity of other gpi anchored proteins. Together these data targets. Wesuggest of have found that no evidence set that Notum3 inhibits the function of limited Glypican4 (Knypek) in a has Notum3 that suggests phenotypes overexpression Notum3 of analysis Our that Notum3actstorestrictthespreadofWntssignalingfromroofplate. of function studies that Notum3 functions in the patterning of the dorsal neural tube. consistent with Wnt/beta-catenin inhibition. In Weparticular, we have propose shown through loss and gain Wnt/beta-catenin. Notum3. of Ectopic expression of Notum3 induces stage-dependent perturbations of of development inhibitor overexpression inducible allow to Tg(hsp70:notum3), line, potent transgenic stable a created have a as with function synergize can the well-established Wnt/beta-catenin inhibitor Dkk1 and to suppress posterior body formation. can overexpression We Wnt8 of it effects the antagonizes that Notum3 Specifically, demonstrates Notum3 of called homologues these of one of expression the regulates signaling We have identified multiple zebrafish several glypicansbygpicleavage. work has demonstrated that a mammalian Notum homologue can similarly induce the release of Wnt, Wingless, by prototypical shedding the the of glypican signaling Dally the and antagonize Dally-like to found from was the Notum cell-surface. called hydrolase Biochemical signaling pathways including several Wnts, of in ligands both for invertebrates co-receptors and vertebrates. as serve that A anchor secreted (gpi) alpha/beta- glycosophosphotidylinositol a via surface cell the to linked proteoglycans sulfate heparan of extracellularly.class a are Glypicans and intracellularly both signaling Wnt of regulation tight requires development Proper embryo. developing the in processes numerous for crucial are proteins, secreted of family large a Wnts, Children’s Pediatrics, of Department Medicine, Memorial Research Center, of Chicago, USA School Feinberg University Northwestern J. Topczewski, G.ParkerFlowers,andJ.M.Topczewska signaling Wnt/beta-catenin regulating loop feedback negative the of element an is Notum-homologue Signaling notum homologues. We determined that Wnt/beta-catenin 114 . Overexpression notum3. Drosophila Drosophila 185

Posters Posters 186 in thezebrafish. PSM the of segmentation underlies signaling Wnt/β-catenin oscillator.Therefore, Notch the to periodicity of Notch activity by controlling the expression In signaling. Notch to responsive summary,is canonical that Wnt signaling PSM is required the to maintain of the identity area of we PSM the cells; activity,Wnt determines initiates Wnt Wnt that decreasing established transiently By transcription. nrarpa regulates the positively sustains signaling and effect inhibitory the Wnt/β-catenin of that evidence provide we Furthermore, duration PSM. the in activity the Notch of presomitic oscillation limits protein the Nrarpa of turnover from fast a somites of formation ankyrin periodic the controls mesoderm (PSM); it involves Notch, Wnt and FGF signaling pathways. Nrarpa (Notch clock segmentation The Children’s Medicine, of School Feinberg Memorial Research Center, University ChicagoUSA Northwestern Pediatrics, of Department J. M. Topczewska, A. Lacrosse,G.ParkerFlowers,J.Topczewski Role of WNT/β-CATENIN signallingduringsegmentationofthepresomiticmesoderm 115 repeat protein ) is both a target and an inhibitor of Notch signaling. We propose that Wepropose signaling. Notch of inhibitor an and target a both is a) of nrarpa, and permits cells to respond Signaling regulated Child Health and Human Development;Human and Health Child cellular functionsandthemodulationofinductiveinteractionsinembryo. embryos. We are current investigating the function of gastrula. at localizations that ventral showed results preliminary Our and expression margin the by replaced is which maternally, of cranial NC into head cartilage in both fish and frog embryos. The embryos. frog and fish both in cartilage head into NC cranial (NC), of crest differentiation terminal neural for required inis it that later showed knockdowns MO and by function of ectoderm loss and early the in predominantly expressed zygotically is in zebrafish: in three are There manners. kinase-independent and kinase-dependent both in PAK4, modifications, subsequently change cell migration. These effects correlate with the inhibition of microtubule and formation fibers stress including dynamics cytoskeletal regulate GTPase, small myosin of Rho-class (PAK4),of 4 effector kinase an p21-activated with physically interact proteins Inka unconventional an PCNS, protocadherin the MyoX, andanovelproteinwenamedInka(formerlyInca). several, molecule, identified adhesion and novel NC, a in out including expressed carried genes TFAP2a-target we function for this screen understand microarray-based further Toa head. embryonic the in tissues other and a critical element in the induction, migration and differentiation of cranial NC cells into skeletal is the transcriptional activator protein AP2a (TFAP2a). We and others have shown that TFAP2a is border. This is accompanied by the expression of numerous transcription factors, among which by interactions among neuroectoderm, the epidermis and paraxial mesoderm at the neural plate including the zebrafish. At the end of gastrulation in vertebrates, the neural crest (NC) is induced Complex and linked inductive interactions pattern the early embryo across the animal kingdom, Research Institute, NationalInstitutesofHealth, Bethesda, U.S.A 1,3,4 T. T. Luo The signalingcomponentInkalinkscytoskeletal dynamics topatternformation Signaling rga i Gnmc o Dfeetain Lb f oeua Gntc, ainl nttt of Institute National Genetics, Molecular of Lab Differentiation, of Genomics in Program 1 , S-K.Hong inka1a,inka1b 2 , Y. Xu 3 , T. D.Sargent and . Each has a unique expression pattern. The pattern. expression unique a has Each inka2. is involved in the early axis patterning of zebrafish of patterning axis early the in involved is inka1b 2 Medical Genetics Branch, National Human Genome Human National Branch, Genetics Medical 4 inka1b, and the possible links between its 116 inka1b gene is expressed is gene gene inka1a genes inka 187

Posters Posters 188 is regulation cytoskeleton mutant. and theirdifferentiationintolensfibercellsarecompromisedintherw341 that suggesting cells, lens disorganized in within fibers accumulated actin that abnormally revealed labeling were Phallodin surface. lens entire the into cells progenitor lens of mislocalization the in resulting disrupted, is epithelium lens anterior of integrity the later but the in normally proceeds differentiation cell lens Wethat dpf. found lens development. differentiation. To elucidate this mechanism, we identified zebrafish mutants showing defects in in which to study the regulation of cell shape change and epithelial polarity associated with cell and cover the core of old lens fibers like the layers of an onion. Thus, lens provideslens asphere and goodstart model to differentiate into secondary lens fiber cells, which subsequently elongate the of zone equatorial the at cycle cell the from exit cells progenitor Lens cells. progenitor lens sphere lens the mitotic of half of posterior consists the differentiate into primary initially in lens fiber cells, whilecells anterior which epithelial cellsepithelial are maintained as mitotic epithelium, development, lens single During of cells. progenitor sphere to a andinvaginates vesicle, lens primordium, the theretinal form overlaying ectoderm in specified is placode Lens Technology, Okinawa, and Science Japan of Institute Okinawa Unit, Neurobiology Developmental T. Mochizuki, M. Yamaguchi andI.Masai Zebrafish mutantshowing defectsinlensepithelialintegrityandfiberdifferentiation 117 rw341 rw341 is rw341 one of these mutants and shows a protruding lens phenotype after 4 mutant lens cells. These data suggest that the integrity of lens epithelium mutant until 3 dpf, 3 until mutant rw341 Organogenesis degradation. These data suggest that lens fiber differentiation is compromised in the through theubiquitinproteasomesystemisimportantforlensfiberdifferentiation. markedly elevated in the was level their and cells, lens differentiating wild-type of nuclei in accumulated were proteins the that found We in the transparency. their However, for nuclei as such organelles subcellular their lose and fibers lens volvox ( we Previously,identified zebrafish mutantsprocesses. showing defects in lens development. Here, wedifferentiation report one of them, cell with associated change shape cell regulates that lens old of core the fibers likecover the layersand of anelongate onion. Thus,subsequently lens providesand a goodsphere, model tolens investigate thethe mechanism of zone equatorial the at cells fiber lens into differentiate to start cells progenitor lens development, During cells. the lens vesicle, a sphere of single epithelium, which initially consists of mitotic lens progenitor Lens precursor region is specified in ectoderm overlying the neural retina and invaginates to form Developmental Neurobiology, OIST, Okinawa, Japan F. Imai, A. Yoshizawa, I.Masai Ubiquitin proteasomesystemisessentialforlensfiberdifferentiation Organogenesis vov vov mutant, lens cells fail to fully elongate into lens fibers, and their nuclei do not undergo ). Differentiating lens cells normally change their cell shape into highly elongated highly into shape cell their change normally cells lens Differentiating ). vov mutant gene encodes a component of proteasome. Poly-ubiquitinated proteasome. of component a encodes gene mutant vov mutant. Taken together, these data suggest that protein degradation 118 vov mutant. 189

Posters Posters expression innotochordcellsorthesurrounding sheathcells. by means of cell transplants determine if the notochord problem is due to the lack the big notochord cell of vacuole. Weautonomous are in the process to analyze Limp2a expression pattern and vacuoles, therefore we concluded that Limp2a participate on vesicular fusion necessary to form numerous with and smaller are cells mutant Notochord provided. is food if even die, larvae the over- neuroectoderm by when dpf 9 – caused 7 till persist it not dpf 2 at apparent is became phenotype notochord The this proliferation. Apparently on. later disappear that hpf 28 – 20 at sn hsooy n eeto mcocp w aaye te hntpc hrceitc o the of characteristics phenotypic the analyzed limp2a we microscopy electron and histology Using the sametimeavaluabletoolforstudyingvacuolenotochordformation. is a useful model to understand the underlying molecular problems in the AMRF disorder and at 190 limp2a mutant larvae zebrafish in also essential for fish larvae swimming behavior as it support somite contraction. We report that Evidently,form. is straight it and notochord, elongated the characteristic larvae the maintaining cells, each bulky containing and a single packed swollen tightly vacuole. with In filled zebrafish thetube notochordelongated keeps thean trunkis rigid it notochord The skeleton. bone the by replaced eventually body.structure developing transient the a of is shape It the maintaining On embryos, from all vertebrates, the notochord is a rod like organ that supports the body trunk Nacional Universidad Celular, Fisiología de Instituto Autónoma Molecular, deMéxico, UNAM, Mexico Genética de Departmento I. Guerrero-Garduño,J.L.Ramos-BalderasandE.Maldonado zebrafish development during formation notochord and brain in LIMP2 protein membrane lysosomal the of Therole ass cin ycou-ea fiue AR) a ua dsre, hc cmie focal combines which disorder, human a (AMRF), glumerulosclerosis andprogressivemyoclonusepilepsy. failure Thereforezebrafishmutantlimp2a it myoclonus-renal mutated When Action Beta-glucocerebrosidase. causes for receptor lysosomal membrane the is Limp2a impairing, atthesametime,swimmingbehavior. vacuoles are not formed properly and in consequence abnormal shaped notochords were found, 119 hi1463 mutant. We found abnormal protruding lumps in the hindbrain and midbrain region hi1463 , that lacks expression of the of expression lacks that , gene, notochordal gene, limp2a Organogenesis hi1463

D) a b a iprat atr n eemnn T determining in factor important an be may (Ds) substantial inhibition of embryonic T 1 that zebrafish embryos are dependent on T on dependent are embryos zebrafish that obnd ncdw o tp 1 n tp 2 eoiae svrl dsub embryonic disturbs 2 severely deiodinases 2 type and V. M.Darras 1 type development inzebrafish of knockdown Combined Organogenesis or T or on the maternal TH deposit in the egg yolk, which consists predominantly of tetraiodothyronine effect on the expression of a wide variety of TH-responsive genes. In teleosts, early embryos rely stimulatory their via development vertebrate in role important an play (THs) hormones Thyroid Universiteit Leuven, Belgium mro, h to eoiae cpbe f T of capable deiodinases two the embryos, important when TH levels remain within the normal range, it may be crucial under depleted under T crucial when case the be is as may status thyroid it very range, be normal to the seem within not remain does levels TH D1 when although important that show results These ear. inner the of the structures fertilization of post hours 31 At batch. observed abnormalities included per extremely curved bodies, posterior truncation and the embryos absence abnormal morphologically 20-25% of only delayed development more than D2 knockdown alone, but also resulted in the occurrence activation in developing zebrafish embryos. Importantly, combined D2 knockdown of contrast, for D1all three morphological parameters andscored, In suggesting that D2 is D2the major contributor to progression. not TH embryos) injected developmental MO control to affect (relative delay developmental clear to a in resulted seem knockdown not did alone via D1 estimated of was Knockdown index. pigmentation development and angle embryonic trunk to of head length, vesicle speed otic indices: the morphological both. three on of treatments combination these a or of D2, impact D1, The against (MOs) oligonucleotides injection morpholino via antisense expression of their inhibiting by relative development their embryonic determine zebrafish to for tried Weimportance fertilization). post hours (8 tested stages earliest the from the thresholdlevelnecessaryfornormaldevelopment. aoaoy f hraetcl ilg, eatet f hraetcl cecs Katholieke Sciences, Pharmaceutical of Department Biology, Pharmaceutical of Laboratory and Neurobiology; and Physiology Animal Endocrinology,ComparativeSection of Laboratory 4 . Therefore activation of T of activation Therefore . 1 , C.N.Walpita 1 , A. D.Crawford 4 to 3,5,3’-triiodothyronine or T or 3,5,3’-triiodothyronine to 3 4 production by D2 is inhibited. The present study suggests study present The inhibited. is D2 by production to T 3 conversion reduces intracellular T 4 uptake and its subsequent activation to T to activation subsequent its and uptake 3 rdcin D ad 2 ae led expressed already are D2, and D1 production, 2 3 dpnet eeomn. n zebrafish In development. -dependent 3 by iodothyronine deiodinases iodothyronine by 120 3 availability below 3 and that and 191

Posters Posters 192 their multipotentialprogenitorsandfurtherdifferentiation. of maintenance the factor,controls transcription which common Sox9b, a on rely pancreas the Our data indicate that, as suggested by their closely related embryonic origin, both the liver and hepatic andpancreaticdevelopment(FGFs,BMPs,Wnt)iscurrentlyinvestigated. for crucial be to known signals various by sox9b of regulation The formation. ducts pancreatic generates endocrine cells. We also present some evidences of the implication of sox9b Surprisingly,pancreas. and embryos mutant sox9b in normal appears bud in pancreatic dorsal the hepato- altered an specified. display mutant the in observed those to processes defective similar from mutants resulting development hepatic correctly sox9b of addition, maintenance In are differentiation. failed poor liver both to and to the state due progenitor the apparently and grow pancreas not does ventral pancreas the ventral the However, both embryos, mutant sox9b In Its expressionisafterconfinedtothewholehepato-pancreaticductalnetwork. that reveals embryos progenitors. bud pancreatic ventral the in then zebrafish and hepatoblasts, in expression the by followed of cells), endocrine of region comprised (exclusively bud pancreatic dorsal hepato-pancreatic the in detected first the is sox9b in analysis expression The and larvaeloss-of-functionforasox9zebrafishorthologue,sox9b. role for this gene has been assigned yet in this structure. Here we report the analysis of embryos progenitor cell pool. In the adult, sox9 expression is restricted multipotent to the pancreatic pancreatic ducts though the no of maintenance the for required and in expressed be to shown was in mammals, like liver the zebrafish in primordium, hepatoblasts give while, birth to hepatocytes and cells biliary cells. In the endocrine mouse In embryo, sox9 and ductal acinar, to anlagen. rise gives ofmultipotential anlage endodermal pancreas ventral the neighboring differentiation in and appearing specification involves progenitors development liver and Pancreas Molecular BiologyandGeneticEngeneeringUnit, Giga-Research/University ofLiège, Belgium B. Peers, I.Manfroid,F. Naye,N.Detry, andV. Von Berg Dissecting theroleofsox9binhepato-pancreaticdevelopment inzebrafish 121 Organogenesis kidney developmentanddisease. vertebrate embryogenesis, and will likely provide useful insights into both normal and defective during formation nephron drive that networks molecular the assemble to framework powerful a establish studies two These Irx3b. and like proximal factors transcription later-acting two by into segments tubule refinement distal broad dynamic two and into sequential by patterned followed initially RA, is by IM domains the whereby model a suggests analysis our together, identity.segment Takendistal a acquire to is progenitors renal of fate default the Irx3b, RAand from signals of absence the in that suggests This segment. distal second the of markers express which nephrons develop Irx3b and is RA nephrogenesis for doubly-deficient in embryos function Furthermore, Irx3 conserved. that Irx3 evidence for providing role thus pronephros, reported amphibian recently the expansion a in acorresponding recapitulates phenotype andprevent this Interestingly, fates. segment proximal tubule to of distal needed first is the Irx3b of factor identity transcription the Iroquois confer the that discovered has strategy This are groups. progenitor we renal various RA, the to of unique downstream genes of pathways knockdown morpholino-mediated molecular performing the delineate To fates. segment proximal of is nephron RA, resulting exogenous entirely comprised is nephron resulting the and identity the with rostral a adopt progenitors renal IM the treated and embryos in identity Conversely, fates. caudal segment a distal of adopt solely progenitors comprised renal IM the absent, progenitors When is domain. renal rostral RA the of specifying for examination requisite was An RA that nephron. found embryos mature RA-deficient in the of domains segment presage that intermediate separated the initially are in progenitors populations distinct kidney into markerssubdivided progressively get then that domains caudal and rostral IM into renal arise. nephrons of the which expression from the (IM) mesoderm examined we origin populations, the determine Toactivity. segment (RA) acid of retinoic on dependent is pattern and segment nephrons, mammalian this to that fashion similar a in organized are that segments distal two and renal the zebrafish among embryo is a good model of nephrogenesis, as it forms nephrons with identities two proximal segment establish that progenitors during pathways nephron formation remain developmental poorly understood. TheWeof segments have cells. previously specialized shown of that comprised epithelial is and kidney the of unit functional the is nephron The Center forRegenerative Medicine, MassachusettsGeneralHospital, Boston, USA transcription IRX3B R. A. Wingert, C.E.McDeed,Z.Kostun,R.Ethier, and the A. J.Davidson and acid retinoic byfactor togenerate proximo-distaldomainsinthezebrafish pronephricnephron patterned sequentially are progenitors Renal Organogenesis 122 193

Posters Posters carrying the -4kb sequence from the start codon of codon start the from sequence -4kb the carrying vector study,reporter GFPthis the In muscles. cardiac and slow adult and embryos torafugu in 2007). al., et (Ikeda study, we examined transcriptional regulation in the expression of of patterns expression complex such in involved mechanisms different muscle-fiber types such each as of slow, expressions fast and stage-specific cardiac developmental ones. However, and specific the details of molecular genome, vertebrate In protein. muscular structural and contractile major the myosin, of subunit large a encodes (MYH) gene chain heavy Myosin Faculty of Agricultural andLifeSciences, The University of Tokyo, Japan S. Kinoshita, Y. Sugano, Y. OnoandS.Watabe be a key regulatory factor. The GFP expression by factor.expression GFPregulatory The key a be to reported been has signaling hedgehog fibers, muscle slow superficial of differentiation the In specific to fast muscle myosin, indicating that these correspond to superficial slow muscle fibers. anti-engrailed GFP-expressing cells were All with stained cells. with F59 antibody pioneer specific stained tomuscle slow muscleor myosin but not notfast with F310medial antibody fastfibersin were not are they cells that medial suggesting antibody, GFP-expressing of and cells. muscle muscle pioneer slow skeletal muscle Myotomal to superficial addition fertilization. muscle, offast post days consists 1-2 embryos zebrafish at embryos transgenic of muscles skeletal myotomal and cardiac in observed was GFP the of expression The embryos. zebrafish control. These results indicate that the -4kb sequence the from the in start codon that of to compared inhibitor,as signaling hedgehog a cyclopamine, with treated embryos region of 62b. Then we created deletion mutants in the -4kb region of region -4kb the in mutants deletion created we Then 62b. of region MYH torafugu of codon start the from sequence upstream study,-5kb this In embryos 2008). al., zebrafish et in (Elworthy muscle slow superficial in expressed isoform MYH an encodes Smyhc1 conserved amongdifferentfishspecies. for gene expression specific to superficial slow and cardiac muscles, and its regulatory activity is 194 myosin heavy in thetorafugu region chain gene,MYH thepromoter of analysis functional and Identification MYH various transgenicapproachesarepossible. zebrafish in vertebrates, among size genome smallest the having rubripes, -1kb to-4kbareinvolvedinthetranscriptionalregulationof MYH the progressively decreased -4k~-1kb rate of and GFP expression, -4k~-1.5k suggesting that not -4k~-2k, a single of sequence but deletion several regions distributed the from hand, other the expression in slow and cardiac muscles with the same activity as that having the intact -4kb. On their functions. Unexpectedly, the reporter construct lacking the conserved region induced GFP 123 M5 M5 is one of the torafugu the of one is n zebrafish and M5 , involved incardiacandsuperficialslow musclespecificexpressions hybridization and RT-PCR previously showed its specific expression RT-PCRspecific and its hybridization showed situ previously In ee oprd y AA porm yedn oe conserved one yielding program, LAGAN by compared were smyhc1 identified from the genome database by database genome the from identified MYHs forms multiple gene family,gene MYHmultiple forms tissue- and MYH MYH M5 promoter was apparently reduced in reduced apparently was promoter M5 was constructed and injected into injected and constructed was MYH lead to the formation of formation the to lead MYH MYH M5 . s are still unknown. In this In unknown. still are s M5 from torafugu MYH Organogenesis MYH where rerio Danio screening silico in M5 and examined and M5 is sufficient Takifugu renal birthdefectsanddisease. during kidney development and may provide insights into the arise molecular mechanisms podocytes that cause how of understanding better a provide data Takentogether,these mesoderm. partially redundant fashion, a in downstream act factors of transcription these RA, that suggesting to specification podocyte induce of loss podocyte complete a cell to Double fate from progenitors. the podocyte intermediate in expression its for signaling RA following knockdown of the forkhead transcription factor observed was phenotype Asimilar post-fertilization. hours 24-48 between decline progressive of Knockdown of induction the for Suppressor-1a ( gastrulation of end the at window podocytes small from the intermediate mesoderm. a Expression of the during transcription factor essential be to found of was (RA) number acid Retinoic a formation. podocyte in identified involved are have that pathways developmental we key kidney, pronephric two-nephron simple asize-selective its with via model, a as blood the filters and governing podocyte kidney formation during renal development pathways are the developmental poorly understood. The Using podocytes. zebrafish called of cells epithelial specialized unit contains that sieve functional the is nephron The Center forRegenerative Medicine, MassachusettsGeneralHospital, Boston, USA A. Davidson, Z.Kostun,M.Grimaldi,R.Wingert, R.Selleck,L.O’Brien. mesoderm Retinoic acid acts upstream of WT1A and FOXC1A to specify podocytes from the intermediate Organogenesis led to an early reduction in podocyte progenitor number followed by a by followed number progenitor podocyte in reduction early an to led wt1a , mlctd n ooye ifrnito, s eedn o R signaling. RA on dependent is differentiation, podocyte in implicated wt1a), foxc1a, which is also dependent upon knockdown led knockdown wt1a/foxc1a 124 Wilms’ Tumor 195

Posters Posters junctions that mediate intercellular communication. Null mutant of this gene, namely gene, this of allele leo mutant Null communication. intercellular mediate that gap-junctions of component a is Connexin gene. connexi41.8 on mutation the by caused is phenotype this and one pattern, stripe is of spite in pattern zebrafish spot has study.mutant pattern This the of for models good the mutant of pattern pigment a leopard, unclear. remains formation pattern of the mechanism molecular However pattern. stripe make cells pigment the consequently body, fish on wave reaction-diffusion like behave xanthophore and melanophore cells, pigment that Weproposed animals. have of pattern skin is nature in seen features beautiful most the of One Nagoya University, Japan other hand allele leo allele hand other phenotype; smaller spots than that of allele leot1 and reduced number of melanophores. On the of leo change thestripetospotpatternbutalsowidthofstripe. to only not factor key a is connexin that indicate data Our pattern. pigment unique to lead can promoter cell-specific pigment of control the under mutants its or connexin of overexpression 1 of pattern in zebrafish. Allele leo Allele zebrafish. in pattern of 196 M. Watanabe Functional analysisofzebrafish connexin41.8forpatternformation Graduate School of Frontier Biosciences, Osaka University, Japan; 125 tw28 is waved but the number of melanophore is as same as that of WT. Here we show that t1 , shows clear spot pattern. Dominant negative mutations in this gene induce variations 1 andS.Kondo tw28 , I31F substitution, shows weaker phenotype than that of leo of that than phenotype weaker shows substitution, I31F , 1,2 tq270 ,

which has I202F amino acid substitution, shows severer shows substitution, acid amino I202F has which 2 Graduate School of Science, Organogenesis tq270 ; stripe ; is negativelyregulatedbyNotchsignaling. intestinal cell proliferation and regulate the differentiation of enterocytes and goblet cells which inhibit to suppressor tumor a as function may KLF4 homologue mammalian its like KLF4a that demonstrate results our Overall, signaling. Notch by regulated negatively was KLF4a revealing DAPT,a by inhibited was signaling Notch which embryos in higher was of level expression Additionally,embryos. the wild-type in than morphants hpf 120 and in less were numbers cell goblet that showed cells goblet within mucins acidic mn ai sqec sae 6.% iiaiy ih mammalian with similarity 65.8% shares sequence acid amino Zebrafish of MolecularandCellular Biology, National Tsing HuaUniversity, Hsinchu, Taiwan, R.O.C. Zebrafish Krüppel-like factor 4a functioning like mammalian KLF4 tumor suppressor is tumorsuppressor KLF4 like mammalian IC. Li 4afunctioning negatively regulatedby Notch signalinginembryonic intestinalcelldifferentiation factor Krüppel-like Zebrafish Organogenesis 1 on the differentiation of enterocytes. Moreover, results from both whole-mount epithelium. intestinal of proliferation the Decreased inhibits 102hpf KLF4a and that hpf indicate 72 results These in embryos. cells intestinal type wild stratified in epithelium intestinal columnar of layer pseudo single with compared as morphants KLF4a of appearance the analyses ultrastructural showed addition, TEM In embryos. by type wild in that than higher significantly were ubr o PopoHsoe 3 tie itsia cls n 4 hpf 74 in The cells tract. intestinal gastrointestinal stained of H3 morphogenesis Phospho-Histone in of function numbers its analyze to used was knockdown oligonucleotide morpholino antisense specific KLF4a hpf. 48 liver,from pancreas exocrine and expressed in the intestine at 36 hours post fertilization (hpf) and can be detected in the intestine, Institute of Cellular and Organismic Biology, Organismic Cellularand of Institute AcademiaSinica, Taipei, Taiwan, R.O.C.; agr2, 1,2 and S.-P. L.Hwang which is expressed in goblet cells in the intestine, and alcian blue staining that can stain cDNA encoding a zinc finger transcription factor was cloned anddeduced was cloned factor transcription finger zinc a encoding cDNA KLF4a expression in 75 hpf 75 IFABPin expression 1 morphant embryos indicates KLF4a may mediate may KLF4a indicates embryos morphant KLF4a 126 KLF4. KLF4a -secretase inhibitor,γ-secretase in situ hybridization trs o be to starts KLF4a opat embryos morphant 102 hpf 102 KLF4a 2 Institute KLF4a 197

Posters Posters GBM attachmentsandcouldleadtonewtherapiestreatglomerulardiseases. edemic the rescues partially inhibitors phenotype. Taken calpain together, these data have identified a novel pathway with used to retain podocyte- embryos knockdown PGAF of treatment idea, this of support In GBM. the from processes foot of detachment subsequent and adhesion in involved proteins of cleavage the PGAF.in with results interaction This unregulated its of loss is the to activity due calpain embryos, knockdown the in that hypothesize We GBM. the to podocyte the link help that proteins of cleavage the through adhesions cellular of detachment the promote calpains Active protease. cysteine calcium-activated a calpain, with interacts and found that PGAF is a direct target of Wilms’ tumor suppressor-1, a podocyte transcription factor, detachment the of reveals numerous embryos podocyte knockdown foot from processes glomeruli from of the analysis underlying isexpressed microscopy GBM. Electron Subsequent malfunction. analysis PGAF embryos. kidney of sign zebrafish a edema, induces knockdown in morpholino and podocytes GBM in exclusively almost the to attachment podocyte maintaining for 198 have We mammals. to compared expression gene identified a novel andcoiled-coil protein, PGAF (Podocyte-GBM function, structure, in conservation filtration the compromises Detachment barrier leading diseases. to proteinuria and eventual kidney kidney failure. Zebrafish podocytesvarious show remarkable of hallmark a is detachment process Foot filtrate. the into molecules small and products waste of passage the allowing but circulation in proteins plasma retaining for important is that barrier filtration the form together The fenestrated endothelial cells of the capillaries, the GBM and the interdigitated foot processes the underlying glomerular basement membrane (GBM) and envelope the glomerular capillaries. of podocytes. These specialized epithelial cells form interdigitated foot processes that adhere to differentiation proper the requires barrier filtration glomerular kidney the of development The School, Medical Boston, USA Harvard Hospital, General Massachusetts Medicine, Regenerative for Center L. O’Brien,Z.Kostunand A. J.Davidson PGAF regulatestheattachment ofpodocytes totheglomerularbasementmembrane 127 attachment factor), that is essential Organogenesis Ishikawa, Japan Kinosh cell proliferationinmedaka. muscle on effect inhibitory an has MSTN endogenous hypertrophy.that not suggest results Our but muscle, (C315Y-MSTN)skeletal MSTN of form hyperplasia dominant-negative induces a of In conclusion, phenotypic analysis of the transgenic medaka revealed that exogenous expression the fibers of whole musclemass. alteration apparent muscle no in the resulting wild-type, of in that Each than smaller medaka. was fish transgenic transgenic in of muscle skeletal the in hypertrophy, not but size and number the analyzed fibers. As a result, and our histological examinations revealed clear evidence offish marked hyperplasia, adult of sections serial made we different, muscle fibers. Since the external appearance of wild-type using and transgenic by fish was not immuno-staining remarkably whole-mount antibody against performed myosin heavy chain. weTransgenic formation, fish showed abnormal muscle alignment of embryonic skeletal the to reveal order In medaka. transgenic the IRES in the cDNAs GFP of C315Y-and between mstn activity located translational element of occurrence the indicated immuno-staining, anti-FLAG by muscle. Green fluorescence, which mimicked exogenous MSTN expression and was visualized skeletal in C315Y-exclusively overexpressing mstn medaka transgenic created Wesuccessfully medaka the by driven expression with (IRES), site entry ribosomal and C315Y-mstn FLAG-tagged harbor to designed medaka in MSTN (C315Y) C313Y-type produce to adenine) (guanine-to- mutation point a introduced we mutant, C313Y-typeMSTN the to corresponding is which of residue corresponding the (C315), %) substituted cysteine to (87 tyrosine of (C313Y) MSTN in reported position the cattle Piedmontese the breed. as and In specify order medaka to mass to obtain a of mutant us muscle regions allowed mature in the increase in similarity an The trait, mice. in phenotypic previously same the causes latipes ) (Oryzias medaka C313Y-typein of MSTN overexpression whether determine to study,sought this we In dominant-negative activity. exhibits protein C313Y-typeMSTN the that implying double-muscling, in resulted MSTN type folding of mature MSTN and is required for full bioactivity. In mice, the in overexpression the of signaling guanine-to- C313Y-portion of the a protein. This is particular cysteine residue there is indispensable for cattle, proper (C313Y) tyrosine a with cysteine critical double-muscle a of substitution a causing mutation transition Piedmontese adenine In mutations. MSTN of caused series be to a shown by been has breeds, cattle domesticated several in fibers muscle of the number in increase an to attributed primarily is which phenotype, double-muscle The cells. target yield to on receptors steps membrane to binds which cleavage of dimer the proteolytic domain, C-terminal mature undergoesbioactive, the that pre-pro-peptide a as synthesized is MSTN of skeletal muscle development and growth. Similar to other members of the TGF-ß superfamily, regulator negative a as act superfamily,to TGF-ß known the is of member a (MSTN), Myostatin . Sawatari E. number ofthemusclefibers increased develops myostatin of form dominant-negative overexpressing medaka Transgenic Organogenesis 1 Aquaculture, Mie, Japan; Bioscience and Biotechnology Center, Nagoya University, Japan; 4 1 . Seki R. , 4 Graduate Schoolof Agriculture, Kyoto University, Japan 1 T Adachi T. , 3 Department Department of Health Science, Ishikawa Prefectural Nursing University, 1 H Hashimoto H. , 1 S Uji S. , GP cNs ond y internal by joined cDNAs (GFP) II hrgfp 2 Y Wakamatsu Y. , DA Te rngn was transgene The cDNA. mstn 2 National Research Institute of 128 -actin regulatory regions. regulatory ß-actin 1 T Nakata T. , mstn cDNA of muscle of 3 M. , 199

Posters Posters 200 that at those from considerably differ patterns evolution. expression for vertebrate machinery core throughout the conserved that is indicating OT, adult pluripotency the neural of PZ the in expressed are genes These 1 andBmi1. We also analyzed the expression patterns of the pluripotency-associated markers in thejuvenilebrain. in the marginalmost regions of the proliferation zones (PZ) of the adult brain but are not detected subset of long-term label retaining cells that are likely to be the OT stem cells. They are located CldU) at different intervals in juveniles and adult fish. This strategy lead to the identification of a To characterize further this region, we performed two long pulses of nucleoside analogs (IdU and actively dividingprogenitorslocatedattheperipheryoforgan. of population a from originating cells progenitor of rings open of additions successive by grows optic tectum (OT). We previously demonstratedthat this cortical structure of the dorsal midbrain To this aim, we use an ideal model found in the midbrain of Neuronal the medaka forebrain. unknown. the to restricted mammalian brain. is the of regions many in mammals life-span throughout occurs teleost in in production neurogenesis Adult undocumented. remains thus diversity their and detail in described been have cells stem neural Veryadult few U1126 INRAgroup, UPR2197DEPSN, InstitutFessard, CNRS, GifSur Yvette, France A. Alunni, J-MHermel, A. Heuze,F. Jamen,F. Bourrat,J-S.Joly Characterisation ofneural stemcellsinmedakafishmidbrain powerful modeltostudythediversityofneuralstemcells. 129 a molecular mechanism specifies an adult stem cell niche. progenitors have similar characteristics to the mammalian neural stem cells remains embryonic stages. This provides evidence provides This stages. embryonic Fish tectum thus represents a new oevr te dl gene adult the Moreover, fish Organogenesis Oryzias latipes, Oryzias the Sox2, Whether non- Whether Musashi into themolecularstructureandexpressionpatternsoflowervertebrateTnnIgene. insight more provide may above, noted as results, the that suggested is it model, system our as 16 hpf, but then extended its expression in craniofacial muscle after 48 hpf. Thus, using zebrafish a novel tnni isoform of zebrafish was expressed exclusively in heart during early cardiogenesis at which they were after identified(hpf), aspost-fertilization slow-h and16 fast-muscle-specific,until respectively.somites the Interestingly,in detectable tnni-HC, not were transcripts tnni2 and transcription-polymerase reverse chain addition, reaction (RT-PCR) In respectively. and vertebrates, whole-mount in other situ hybridization of demonstrated that TnnI2 zebrafishmuscle tnni1 fast and TnnI1 muscle slow the of homologous were cloned we TnnI2 and TnnI1 zebrafish the and craniofacial). (heart Based tnni-HC on and the tnni2 unrooted tnni1, radial genes: tnni gene zebrafish tree of analysis types of three the of TnnI patterns gene expression among vertebrates, TnnI gene in lower vertebrates. Here, we cloned and characterized the molecular structures and the about cardiac known is little However, in very muscle. skeletal expressed slow and muscle isoforms skeletal fast the muscle, encodes mammals, and birds in identified been has which highly a therefore, is, sensitive and specific marker of myocardial injuryTnnI in acute coronary syndromes. The TnnI gene, Cardiac contraction. muscle striated for switch calciumsensitive a provides filament, thin the on located complex troponin the of constituent a Troponin(TnnI), I Institute ofMolecularandCellular Biology, National Taiwan University, Taipei, Taiwan CY. Fu,H-C.LeeandH-J.Tsai The molecularstructuresandexpressionpatternsofzebrafish troponinIgenes Organogenesis 130 201

Posters Posters Here we focus on focus we Here closely relatedtomammaliansialidaseNEU3. 202 order toconfirmthespecificityofphenotypicalterations observed. for the expression of different marker genes. mRNA rescue experiments are presently ongoing in characterizing the phenotypes caused observed by also whole-mount but alterations phenotypic stronger severe mortality much at 24 to hpf, rise suggesting that gave a complete amounts depletion of injection Higher observed. is develop will system digestive where region the of impairment significant a hpf, 24 Moreover,somites. at misshaped axis, body main the of shortening edema, heart edema, brain of Knock-down using differentmarkers. now plan to characterize the morphant phenotype observed by eye and head development show retardation, Morphants trunk and alteration. tail abnormal phenotypic curvature and significant defective causes somites patterning. knock-down We neu3.1 experiments usingsplice-inhibitingMorpholinos(MO). order of In role respectively.the lens, test to and gut to corresponding areas expression localized spatially and that respectively. The remaining genes, i.e. of them, Twosialidases. human to related ) rerio (Danio Mammalian zebrafish of genes different seven characterized sialidase. and cloned Werecently (NEU4) apoptosis. and adhesion, growth, differentiation, cell including events membrane-associated intracellular cellular important of regulation in as well as catabolism, lysosomal in implicated are sialidases and membrane- (NEU3) plasma (NEU2), cytoplasmic associated (NEU1), lysosomal described: been have sialidases the represent sialidases Thus, glycolipids. sialoglycoconjugates. of degradation the for step essential and andinitial glycoproteins from residues terminally acid of removal sialic the linked catalyze that enzymes of family a are neuraminidases or Sialidases of Biology, University ofPadova, Italy. Department of Biomedical Sciences and Biotechnology, University of Brescia, Italy; *Department D. Gotti,M.Manzoni,G.Paganini,*N.Tiso, G.Borsani, A. Preti,E.Monti,R.Bresciani Preliminary study ontheroleofsialidaseneu3.1andneu4inzebrafish development 131 and neu3.1 neu1 and by injecting 1 ng of MO results in a spectrum of phenotypes ranging from ranging phenotypes of spectrum a in results MO of ng 1 injecting by neu4 are expressed already at early development stages and show temporal show and stages development early at already expressed are neu4 and neu3.1 and neu3.1 neu4, represent the orthologs of the mammalian sialidases . RT-PCRneu4. and in zebrafish development we performed gene inactivation gene performed we development zebrafish in neu4 neu3.1, neu3.2, hybridization experiments demonstrate experiments hybridization situ in neu3.3, in situ hybridization of injected embryos in situ neu3.4 and neu4 is lethal. Four different mammalian different Four hybridization experiments neu3.5, are all more Organogenesis NEU1 and We are currently NEU4, be involvedincontrollingaxonoutgrowthofretinalganglioncells. that found recently We ganglia. to corresponding possibly regions, optic the Outside proceeds, morphogenesis vsp eye As stage. 10-somite the at primordia optic the of region ventral eye. developing the in expression specific displays and domain screening. situ in selected we development, eye in involved genes for Looking University ofExeter, UK; Kudoh irbooy n Mlclr eeis Uiest o Ptsug, USA; Pittsburgh, of University Genetics, Molecular and Microbiology . Andreazzoli M. of region ventral the in expresses zebrafish eye arms/kidins220 of interactor an vsp, of Characterization Organogenesis 1 f ntm ad eeomna Booy nvriy olg Lno, UK; London, College University Biology Developmental and Anatomy of protein ARMS/kidins220, a target of neurotrophin and ephrins. These data, together with the with together data, of These absence ephrins. and neurotrophin of target a ARMS/kidins220, protein aoaoi d Booi Cluae dlo vlpo Uiest’ i ia Italy; Pisa, di Universita’ Sviluppo, dello e Cellulare Biologia di Laboratorio expression becomes restricted to the ventral retina and to the initial tract of the optic stalk. optic the of tract initial the to and retina ventral the to restricted becomes expression 4 , G.Barsacchi expression in the ventral eye region of region eye ventral the in expression vsp 1 G Gestri G. , noe a rti cnann to D dmis n a RING-finger a and domains PDZ two containing protein a encodes vsp 1 , S.W. Wilson 5 Laboratory ofMolecularGenetics,Laboratory NICHD, NIH, Bethesda, USA is expressed in rhombomere 1 and in specific areas of the trunk the of areas specific in and 1 rhombomere in expressed is vsp 2 B D’Orsi B. , 2 , I.B.Dawid 1 M Barilari M. , 5 , L.Dente 1 A Iervolino A. , mutants, suggest that this gene might gene this that suggest mutants, noi 1 specifically vsp in the course of a large scale large a of course the in vsp expression initiate in the in initiate expression vsp 132 1 E Landi E. , 4 col f Biosciences, of School neat wt the with interacts 3 1 eatet of Department M Tsang M. , 2 Department Department 203 3 T. ,

Posters Posters 204 similar to an Morpholino zebrafish. in phenotype a ciliogenesis body, curved reminiscent in to a other with cilia role animals mutants. Unc119bproduces Unc119b a also of displays has knockdown a (MO)-mediated Unc119bcilia phenotype in whether the pronephric tested duct therefore We family members,whichareinvolvedinciliogenesis. done in uptake and the distal tips of cilia are not formed properly.unclear. remains In mammalian role systems, as molecular well as its work The Unc-119 protein is required for normal nervous system development in metazoans, although Dept. ofBiologicalSciences, University of Alberta, Edmonton, Canada D. V. French andD.B.Pilgrim Unc119b RegulatesCiliogenesisandLeft-Right Axis Determination n19 itrcs ih h Ff inlig aha, hptei w ae urnl testing. currently are of we determination and hypothesis left-right asymmetry.ciliogenesis in Unc119b a for role pathway,novel a signalling provides work this Fgf Altogether, the with suggests phenotypes interacts vesicle Kupffer’s Unc119band gland pineal the with coupled result This midbrain- mutants. the Fgf of boundary,reminiscent midbrain-hindbrain that the from excluded is reveals (atoh1a) homologue1a hindbrain the into that reveals analysis Unc119bFurther in morphants. close not does boundary hindbrain Rhodamine-dextran of tube neural open Injection an and phenotype. hydocephalus have Unc119b ciliogenesis, in defects to addition In at Kupffer’s vesicledemonstratesshorterandfewerciliainUnc119b morphants. cilia Unc119bof gene. with Analysis ciliogenesis consistent a are as results acting These event. pineal gland, as The marked by asymmetry. left-right of establishment the in role a indicating looping randomized upstream or parallel of 133 C. elegans in our lab, Unc-119 protein family members have been shown to interact Arl arl13b mutant, and genetic epistasis experiments indicate that , fails to migrate in crx Unc119b morphants, which is a left-right dependent arl13b. In addition, Unc119b morphant hearts are unlooped or displayed mutants in mutants elegans C. have abnormalities of dye of abnormalities have unc-119 unc119b acts either Organogenesis atonal 2 M. A. Moriarty Modelling arrhythmogenic rightventricular cardiomyopathy inzebrafish Organogenesis signaling roles inzebrafishsignaling roles development.’ cardiac Developmental Biology327 zebrafish in E.D.,Martin,and Moriarty, structural M.A.,Byrnes,L.,Grealy,both has M.(2009) ‘Plakoglobin proteins armadillo desmosomal of importance the development andtheusefulnessofzebrafishasamodelfor ARVC. show data These show adecreaseinjunctionnumbersalongwithalteredjunctionalmorphology. numbers of junctions in the heart as a whole. Initial TEM studies in plakophilin 2 morphants also (TEM) showed altered adhesion and cellular morphology tail defects. in Structural analysis their of plakoglobin morphants cardiac using transmission valves electron oedema, microscopy along pericardial technology with - defects reduced enlarged antisense hearts, cardiovascular reduced specific heart bymorpholino beat. with This is embryos accompanied by morphant stage-dependent or plakoglobin anterior in neural resulted and 2 has plakophilin zebrafish of Knockdown 2009). ARVC (Martin for model a is zebrafish in plakoglobin of loss that shown previously have lethality.Weembryonic and instability cardiac to leads mice in plakoglobin and 2 plakophilin important in tissues which undergo routine mechanical stress such as the skin and heart. Loss of disease particularly are and strength, heart and integrity tissue maintaining to integral are These plakoglobin. congenital proteins Several people. a young in and 2 plakophilin in proteins implicated ARVCarmadillo been desmosomal have the including is death sudden and (ARVC) arrhythmias ventricular with Cardiomyopathy associated Ventricular Right Arrhythmogenic 1 Department ofBiochemistry,Department NationalUniversity ofIreland, Galway, Ireland Department of Pharmacology and Therapeutics, National University of Ireland, Galway, Ireland; 1 , E.D.Martin 1 , L.Byrnes 2 , M.Grealy 1 134 et al., et 205

Posters Posters craniofacial muscledevelopment. regulatory putative pathways to understand how four proposed we Therefore, network. regulatory same the in function not in defects muscle cranial the rescue not did 206 by induced those phenocopied pax3-MO by caused defects muscle craniofacial the Although myogenesis. muscle arch during initiation, that suggesting myf5-morphants, of overexpression However, of injection by induced that muscles suggesting cranial in defects the all rescue to of Overexpression mesoderm. head from originate that muscles cranial rectus, for medial sternohyoideus, for required was inferior rectus, Six1a and all pharyngeal arch muscle that development. Although Six1a was also demonstrated necessary cell we Here, and unclear. organogenesis, morphogenesis, still is development muscle in cranial zebrafish during Six1a of roles role However,the differentiation. important plays Six1a factor Transcription Institute ofMolecularandCellular Biology, of National Taiwan University, Myogenesis Taipei, Craniofacial Taiwan the CY. in Lin,W-T. Chen,H-C.Lee,P-H. Yang, H-J. RoleYang andH-J.Tsai Essential an Zebrafish Plays Six1a Factor Transcription The 135 and myod xrsin n ed uce, t i nt affect not did it muscles, head in expression myogenin s iety ontem of downstream directly is myod was unable to rescue arch muscle defects in the in defects muscle arch rescue to unable was six1a six1a distinctly interacts with either six1a s ny novd n ygnc aneac, o its not maintenance, myogenic in involved only is -MO, injection of injection six1a-MO, and six1a that suggesting morphants, pax3 n otoln caifca myogenesis. craniofacial controlling in six1a myf5 or six1a, mrhln (MO), six1a-morpholino myodembryos enabled myod during zebrafish myod xrsin in expression myf5 Organogenesis or mRNA myf5 and tbx1- do pax3 in thethyroidprimordiumanddifferentiatedfollicles. gene. Experiments with embryos and larvae showed that GFP-mCherry is specifically expressed thyroglobulin these zebrafish the In of sequences animals. regulatory living by driven in is GFP-mCherry differentiation fish, and transgenic migration thyroid monitor to us allow line will zebrafish that transgenic a established we addition, In at fivedistinct approaches. overexpression and down embryos in zebrafish expression developmental stages. Positive candidates thyroid-specific are further characterized by using for morpholino knock- genes whole-mount candidate for mouse protocol throughput high a Using mouse. in identified genes candidates the of characterization functional a for model zebrafish the of opportunities unique Wethe development. exploit thyroid to strategy a devised therefore thyroid early Previous studies clearly demonstrated the value in of zebrafish as a model to study early aspects of involved potentially genes candidates development. bioinformatical 1300 various than of more application yielded Subsequent development. approaches thyroid of stage early an at embryos. mouse E11.5This approach in proved successful to endoderm quantitatively and pharyngeal qualitatively characterize gene expression and mesenchyme surrounding primordium, thyroid of transcriptome the analyze to sequencing deep Solexa with LongSAGE combined we differentiation, and development thyroid of regulation the in involved genes novel Toidentify might playanimportantroleduringthyroidorganogenesis. genes unknown other,still that suggests This FOXE1. or PAX8 have NKX-2.1, as such cases development, CH familial of proportion minor a been associated with mutations located in transcription factors known to be involved in thyroid only date, To humans. newborn in of gland ectopy this and/or hypoplasia agenesis, comprises which dysgenesis thyroid of result the is CH In human, congenital hypothyroidism (CH) is a frequent congenital disease. In 85% of the cases, fully differentiated,expressesthyroidspecificgenesandproduceshormones. becomes thyroid the development, of stage this zebrafish. At in aorta ventral the along tissue of and adopts its final bilobed shape in the neckmesenchyme areahypopharyngeal ofthe micein andrelocates manpharynx, orthe formswith an elongatedcontact strand loses primordium the bud derived from the ventral floor of the anterior pharyngeal endoderm.The thyroid is an endocrine gland Aftercomposed of follicular cells that develops from aa primordium migration process, IRIBHM, ULB, Brussels, Belgium E. Maquet,R.OpitzandS.Costagliola Zebrafish asamodeltostudy ofthyroid development Organogenesis yrdzto, e r screening are we hybridization, situ in 136 207

Posters Posters Switzerland; 2 208 homeobox the of role the Neurons: A. E.Fortunato Motor Primary of uncx4.1 development and Somitogenesis 1 ouae Npe, Italy; Naples, Comunale, cell cycleinmuscleprogenitorsatthemargin ofeachsomite. suggest a double role for the Spatio-temporal dynamics of Currently, regulation. extends abnormally from dorsal to ventral within somitic compartments, confirming the negative (nodal), uncx4.1 transcription. Analyses of the shh pathway zebrafish mutants (floating head (flh), uncx4.1 seems to be negatively regulated by shh molecules, suggestingthatuncx4.1activitystepsbackfrommaturingmusclecells. mRNA and MYOSIN protein (fibroblast marker) reveals a complementary distribution of the two precedes that showed hybridizations double differentiation, muscle with relation In the ventro-lateralspinalcord. cells, somitic medial in anymore detected appearance of the first progressively outgrowing primary motor axons. Indeed, after becomes of and restriction stage progressive 5ss the Interestingly, from detectable is level posteriorly andventrallyrestrictedtothepresumptivemyoblastcellsinlaterstages. somite the at expression the particular,In ducts. pronephric and CNS somites, arches, branchial in development embryonic gene The UK. eatet f ellr n Dvlpetl ilg, tzoe olgc Atn or, Villa Dohrn, Anton Zoologica Stazione Biology, Developmental and Cellular of Department ofCellularDepartment and Developmental Biology, University ofPalermo, Italy; 137 and (shh) you sonic ciain n uce rcro cls Mroe, olclzto of co-localization Moreover, cells. precursor muscle in activation myoD is a paired-type homeobox transcription factor, expressed during zebrafish during expressed factor, transcription homeobox paired-type a is uncx4.1 functional characterization is performed by a morpholino mediated strategy.mediated morpholino a by performed is characterization functional uncx4.1 4 Department of Anatomy and Developmental Biology,London,DevelopmentalCollege and University Anatomyof Department 1,2 , T. Kondoh 3 eatet f opoee, cecs I, nvrié e Geneve, de Université III, Sciences Morphogenes, of Department 3 uncx4.1 gene, in guiding outgrowth of motor axons, and regulating uncx4.1 expression, along with functional experimental evidences, , D.Duboule you-too (gli2)) you-too 3 a ventrally projecting CaP motor axons emerge from emerge axons motor CaP projecting ventrally a , N.Holder ciiy niiae ad copne the accompanies and anticipates activity uncx4.1 showed that the expression of the of expression the that showed pathway, indeed shh overexpression abolishes 4 , S.Wilson 4 , P. Sordino uncx4.1 expression is not Organogenesis expression uncx4.1 2 gene uncx4.1 uncx4.1 cyclops Our analysis of the zebrafish genome reveals three reveals genome zebrafish the of analysis Our vertebrate during phosphatases zebrafishas development. Cdc14 of Using function the understand invertebrates. further to aim we understood model, in poorly a studied remains well but been yeast, has fission phosphatases and Cdc14 budding of regulation and role the eukaryotes, among to chromosome segregation, mitotic exit its and cytokinesis. of Although dephosphorylation the and Cdc14 family Cdk1 substrates. is The Cdc14 of family conserved of phosphatases is inactivation involved in this latter the event and thus contributes requires mitosis from exit Specifically, Progression through the cell cycle relies on oscillation of Cyclin-dependent kinase (Cdk) activity. Cell andDevelopmental Biology, HHMI/Vanderbilt University, Nashville, USA A. Clement,L.Solnica-Krezel,K.Gould The cellcycle regulatorCdc14Bisessentialforciliogenesisinzebrafish Organogenesis the processofciliogenesis. cdc14B morphants, strongly suggesting that we have uncovered a novel function for Cdc14B in the in in affected also are defect cilia of types asymmetry other that indicate KV results Left-Right Preliminary morphants. the cdc14B the in for mechanism cilia a the providing of likely length reduced the considerably that is find we however, Interestingly in morphogenesis. unaffected its is is KV so the in cells of number the cycle, cell the during role a plays Cdc14B asymmetry.Although Left-Right establishing for responsible organ when suppressed are expression of expression development. throughout aberrant as well as position liver and pancreas and looping gut shifting, tube heart randomized ubiquitously expressed both oligonucleotides, unexpectedly impairs Left-Right are asymmetry. and Downregulation of identified were isoforms . Here, we report our results on results our report we Here, cdc14B. , the earliest marker showing asymmetric expression. These phenotypes These expression. asymmetric showing marker southpawearliest the , cdc14B function during embryonic development using antisense morpholino RNA is co-injected. In zebrafish, the Kupffer’s vesicle (KV) is the is (KV) vesicle Kupffer’s the zebrafish, In co-injected. is RNA cdc14B

during zebrafish embryogenesis. embryogenesis. Two zebrafish during cdc14B paralogs: cdc14 cdc14B morphant embryos exhibit deficient embryos and embryos deficient cdc14B 138 cdc14A1, and cdc14A2 cdc14B 209

Posters Posters 210 of thesemutantswillbepresentedindetails. some of phenotype The mutants. these in markers intestinal and hepatic pancreatic, various of tissue or on a decrease of enteroendocrine cells in the gut. We pancreatic cells, are several mutants were detected based presently on a reduction of the analysing pancreatic exocrine expression 500 mutagenized genomes were analyzed. While we did not identify mutant lacking endocrine AB/TL (Driever’sfish than More probes. trypsin and glucagon using hybridization situ in by lab) cells pancreatic endocrine cells, beta and enterodendocrine cells. The second screen of was performed on non transgenic mutagenized observation direct the allows which Insulin:DsRed / performed: the first mutagenesis was performed on awere doublescreens transgenicmutagenesis zebrafish2 linedevelopment, pancreas Pax6b:GFP exocrine or endocrine ofselected affecting mutations :i)knock-down identify to order In approaches defects. pancreatic displaying mutants of screening ii) and main genes candidate two using zebrafish in development pancreas Germany Belgium; Liège, Laboratoire de Biologie Moléculaire et de Génie Génétique (LBMGG), GIGA-R, B34, Sart Tilman, F. Naye, M.Voz, J.Schweitzer*,O.Ek,W. Driever*andB.Peers Screening formutationsaffectingthedevelopment ofexocrineandendocrinepancreas The main research theme of our laboratory is to study the molecular mechanisms controlling mechanisms molecular the study to is laboratory our of theme research main The 139 Atiug nwclnsilge Isiu fr ilge , nvriä Freiburg, Universität 1, Biologie für Institut Entwicklungsbiologie, *Abteilung Organogenesis This project is supported by issupported This project an A-STAR/BMRC grant(07/1/21/19/544). skeleton formation:(i)CNCCmigrationand(ii)establishmentofgillcartilagemorphology. possibility that Sost controlled Lrp5 signalling is involved in two independent processes of clusters cranial of cartilage cells that subsequently fail to form the gill cartilage elements. This opens the the region that would form the ceratobranchials. In contrast, knockdown of in a concentration dependent manner. in regions of the ventral head, where CNCCs differentiate to form the gill cartilage structures. expressed cartilage gill are Morpholino the mediated form they knockdown to differentiate of stages, CNCCs either where later head, ventral At the of CNCCs. regions in delaminating the to adjacent directly hindbrain, patterns in the developing head. Early in development, Sclerostin (Sost) are involved in these inhibitor processes. Both genes are expressed in putative adjacent and highly dynamic its and (Lrp5) 5 protein receptor-related (LDL) density-lipoprotein low- co-receptor is frizzled the that signaling show we Here Wnt process. this canonical during steps various that for essential earlier shown been has It structure. cartilage distinct a to rise subsequently differentiate to form the cranial cartilage. Each stream is thereby in committed to give migrate they which in and regions cranial respective the plate invade to ventrally streams neural mesenchymal separate the from delaminate (EMT), transition mesenchymal to epithelial undergoan (CNCCs) rostral cells crest neural cranial these development, the of course the In crest. neural from originate that cells by established is skeleton cranial the of portion major The ofBiological Sciences(DBS),Department NationalUniversity ofSingapore B. Willems, J.RennandC.Winkler skeleton Lrp5 and its putative inhibitor Sclerostin are required for development of the zebrafish cranial Organogenesis lrp5 morphants completely lack cartilaginous structures in lrp5 or sost prevents formation of the ceratobranchials lrp5 and sost are expressed in the dorsal 140 sost results in excess 211

Posters Posters oe n h dfeetain f hrtoe n smttoe el truh nuto of induction through cells somatotrope and thyrotrope expression inthedevelopingpituitary. of differentiation the in role In conclusion, Sox4b is expressed in zebrafish during pituitary development and plays a crucial the effectsobservedinsox4b morphants. in that observe clearly could Finally,we embryos. hpf 48 that showed including markers of levels reduced and expression gsuα with and hpf 33 at in the pituitary anlagen starting at 24 and 26 hpf and in the entire head region including the including region that head show entire We the 48hpf. in at and pituitary hpf 26 and 24 at starting anlagen pituitary the in that found We development. pituitary in family (SOX) HMG-box lactotropes, somatotropes, type in fish expressing somatolactin. types: We investigate the role of Sox4b, a member of the SRY-like cell different six least cell additional an and mammals, in gonadotropes and corticotropes melanotropes, thyrotropes, at of consists adenohypophysis The Bioquímica, Universidad Austral deChile, Valdivia, Chile. M. Muller for Developmental Biology,Developmental for Cologne,of University Germany; a 212 The Zebrafish hmg-box transcription factor sox4b activates pituitary expression of Y. Quiroz specification ofthyrotrope cells Research Institute, San Francisco, USA; transcription factor transcription also mutant this of Expression in decrease a to led proteins. sox4 endogenous on mutant dominant-negative a as (Sox4 domain C-terminal its of LBMGG, Tour GIGA, Liège, Belgium; tshß and 141 a a, , M. Lopez * gsuß expression at 48hpf. but expression colocalizes with colocalizes expression gata2 and prl is downregulated in downregulated is gata2 at 48 hpf. 48 at tshß tshß, a , A. Mavropoulos and gsuß are not affected. We also used a mutant form of Sox4 deleted Sox4 of form mutant a used also We affected. not are lim3 Cter) that is unable to activate transcription and therefore acts therefore and transcription activate to unable is that ΔCter) mRNA colocalizes with the pan-pituitary marker pan-pituitary the with colocalizes mRNA sox4b -positive cells. Furthermore, expression of the zinc finger zinc the of expression Furthermore, cells. gh-positive knock-down leads to a drastic decrease in decrease drastic a to leads knock-down sox4b b Plant Cellular Biology and CATM, Liège, Belgium; a$ d and gh Laboratorio de Biología Molecular de Peces, Instituto de , G. Nica prl expression was not significantly affected; similar to morphants specifically in the pituitary. the in specifically sox4bmorphants We tshß, mRNA, while other anterior pituitary gland pituitary anterior other while mRNA, slß c , P. Motte and gsuß knock-down leads to a decrease a to leads knock-down 2 gata b , J. A. Martial $ UCSF Diabetes Center,Diabetes UCSF Hormone mRNAs in the pituitary of pituitary the in mRNAs sox4b is strongly expressed strongly is sox4b a , M. Hammerschmidt Organogenesis gata2 c and tshβ Institute gata2 lim3 and c , brain ventricle inflation. In sum, these data show that theNa that show data these sum, In for essential inflation. are 6 ventricle and brain 5 FXYD1, subunits, gamma the that suggest functional data of Preliminary amount the Atp1a1. to sensitive highly is area ventricle brain forebrain that show assays midbrain ventricles, and that it synergizes with Atp1a1 during ventricle inflation. Dose response subunit beta the that show we addition, In neurulation. after inflation ventricle in brain for early and development, integrity epithelial for necessary is Atp1a1 that demonstrate we Here 2005). Sive and showed that the alpha subunit, previously gamma. We and beta alpha, - subunits three includes pump This inflation. ventricle ventricle size,resultswithimplicationsfortheunderstandingetiologyofhydrocephalus. before and after neurulation, and that the correct balance of subunits is necessary to direct brain it functions. In the zebrafish, the Na the zebrafish, the In functions. it how or contain secreted is fluid ventricles this how about known brain is little (eCSF) embryonic CSF embryonic of the amounts large Although function. brain for essential is which (CSF), The brain ventricular system is a highly conserved set of cavities, containing cerebrospinal fluid Cambridge Center,Technology;Nine CambridgeUSAof Institute Massachusetts and Research Biomedical for Institute Whitehead J. Chang,L.A.Lowery, H.Sive Regulation ofbrain ventricle inflation Organogenesis is strongly expressed in neuroepithelium surrounding the forebrain and forebrain the surrounding neuroepithelium in expressed strongly is atp1b3a atp1a1, is required for zebrafish brain ventricle inflation (Lowery + K + ATPase ion pump is crucial for initial embryonic brain embryonic initial for crucial is pump ATPaseion + K + Tae ly dfeet roles different plays ATPase 142 213

Posters Posters 1 214 K. Hatta and medaka, and their motionsduringrespirationzebrafish andfeeding in elements skeletal and teeth of structures 3D of imaging X-ray with differentanatomicalcharacteristics. roles by comparing these two teleosts established as model vertebrates that appear to have teeth Functions of pharyngeal teeth in fish have been poorly understood. We hope to elucidate their different with periodically move phase duringrespiration. to found was element skeletal Each during feeding. motions and their study respiration to elements skeletal the of imaging X-ray live performed then We Medaka alsohasteethonupperandlowerjaws. teeth pharyngeal 1000 distributed both than at ventral and more dorsal sides. Dorsal has teeth and ventral medaka teeth face are facing adult each other.to contrast, found In were midline. teeth towards ventral other left each and Right side. ventral the at only teeth pharyngeal ~24 of skeletal elements in adult and larval zebrafish and medaka. Adult zebrafish has relatively large X-ray micro tomography at SPring-8 synchrotron was used to analyze three-dimensional structures Grad. Sch. ofLifeSci. Univ. ofHyogo, 143 1 , K.Fujita 1 , T. Yamamoto 1 , K.Uesugi 2 JASRI/SPring-8 Japan 2 , S.Nakayama 1 , P.K. Moly 1 Organogenesis 1 F. Priller in earlycardiacmorphogenesis Gene expression profiling of zebrafish embryonic hearts identifies molecular networks involved Organogenesis requirements duringheartdevelopment. of impact known the signaling pathways on assess CPC gene function expression. to Thereby we hope profiling their to elucidate their transcriptional specific elucidate employ now to we and addition, candidates In cardiogenesis. prime in verify to started now have we findings, our on Based features. unique its to contribute likely and heart the in up-regulated specifically of are stage the at tissue heart lacking embryos cardiac whole field involutionof reveals thatsample about onereference quartera of allto genes thatexpression fall withingene those categories as these genes are likely to be involved in morphogenetic processes. Comparison of myocardial we are particularly interested in genes that regulate migratory behavior, cell polarity Further,or myogenesis. and adhesion, homeostasis ion heart, the signal of patterning or specification transduction, intracellular extra- with purified include associated These are highly processes. that physiological hundreds or on find morphogenetic particular we profiling heart, the in expression expressed genes gene zebrafish all annotated of transcriptome Out formation. tube heart of whole regulators key and novel identify of to tissue heart embryonic use make we Here, that underlie CPCbehaviorremaintobeunraveled. mechanisms on cellular and based genetic processes exact the these analyses, in phenotypic gain-of-function implicated or been loss- have pathways signaling of couple a Although field. heart the of portions distal within processes migratory extensive and field heart posterior of course the in rearrangements morphogenetic cardiogenesis. In zebrafish, heart tube formation involves the asymmetric involution of the toright subject are which (CPCs), cells precursor The multi-chambered hearts of vertebrates initially arise from two lateral populations of cardiac Delbrueck Center, Berlin, Germany Cardiovascular Research, Max Delbrueck Center, Berlin, Germany; 1 , M.R.Huska 2 , M. Andrade 2 , S. Abdelilah-Seyfried 1 2 144 Computational Biology, Max 215

Posters Posters 216 gluconeogenic program. to expect phosphoenolpyruvate. see a we phenotype that is specific toacids, impaired utilization of those to metabolites,amino but not of the entire glucogenic or oxaloacetate pyruvate lactate, converts from derived that is oxaloacetate pathway Since the of enzyme the is upstream Pck1 controls. most to compared embryos injected in size normal largelyliver a decreased indicate but results development, Preliminary stream. blood the into glucose of release important the is for it where program gluconeogenic the of point end the at functions 6-phosphatase against targeted analyses conducting down are knock we gene embryo, developing Morpholino-mediated the for gluconeogenesis of requirement the study To this hypothesisandgaininsightsintotheregulationofglucoseproduction. this time course of gene expression. Using pharmacological and genetic studies we want to test Energy demand and lactate production of the swimming fish is likely to contribute to the logic of fbp1 show a strong increase around this time point when larvae have hatched of and start onset to swim. the to addition In hpf. 96 and 72 between intestine, and liver in later of differentiation functional At 48 the hpf we detect to expression of prior spot” the “on gluconeogenic organs. glucose to lipids yolk abundant from energy density of lipids compared to glycogen, this early In the early embryo pck1). by (encoded 1,6-bisphosphatase by (encoded Glucose-6-phospatase pathway: gluconeogenic the of enzymes out carried we resistance, insulin and In order to characterize zebrafish as a model for studies on the regulation of glucose metabolism to hyperglycemia inT2DM. Insulin resistance in hepatocytes induces elevated glucose output from the liver and contributes (T2DM). Mellitus TypeDiabetes and 2 Syndrome Metabolic the of part as resistance insulin increase in pandemic a to lead has civilization western in prevalence obesity rising steadily The UCSF, SanFrancisco, USA P. Gut,M.Porte,D.Stainier Characterization andfunctionalanalysisofgluconeogenesisduringzebrafish development 145 pck1 is strongly expressed in the yolk syncytial layer. Considering the higher fbp1 and ) and Phosphoenolpyruvate-carboxykinase (encoded by (encoded Phosphoenolpyruvate-carboxykinase and fbp1) yrdzto epeso suis o tre key three for studies expression hybridization in-situ g6pc in the developing liver whereas pck1 expression suggests a conversion and g6pc Organogenesis , fructose- g6pc), pck1, pck1 is initiated Glucose- pck1. g6pc and process ontumourformation. cellular important this compromising of consequences the explore to and biogenesis, ribosome impaired to response cellular the in isoforms various its and p53 of role the dissect to embryos arrest cycle cell the of levels specifically and markedly upregulated in response to impaired rRNA processing, and the mRNA In to yeast from conserved is function vertebrates. Nol8 Thus, species. rRNA 28S mature of levels reduced in analyses processing rRNA bioanalyzer defective demonstrate and to blot Northern alongside experiments labeling metabolic employed gene that has been shown to be required for ribosomal RNA (rRNA) processing in yeast. We have y ae lo akdy ne-eeoe ad rnoail atlg dvlpet s severely is in mutation responsible The development cartilage craniofacial and disrupted. under-developed markedly also are eye in intestinal epithelium the fertilisation post days 5 At morphogenesis. organ endodermal in involved genes G. J.Lieschke eet i rbsm boeei udri te opooia caatrsis f zebrafish a A.P. of Badrock characteristics morphological the development mutant,setebos,witheye, intestinalandcraniofacial abnormalities underpin biogenesis ribosome in Defects Organogenesis 1 protein, p53. The ribosomal protein L11 has been shown to be a key player in this process, setebos this in player key a be to highlighting theneedtounderstandextra-ribosomalfunctionsofribosomalproteins. shown suppressor been tumour has L11 the protein of ribosomal activation The through p53. arrest protein, cycle cell or apoptosis in the resulting stress’ yet, ‘nucleolar to leads As biogenesis ribosome of disruption mammals, cells. In understood. growing incompletely in process consuming are proliferation energyand growth cell of regulation the to biogenesis ribosome link that mechanisms most the is biogenesis Ribosome Medical Research, Royal Parade, Parkville, Australia of Biochemistry and Biophysics, University of California, San Francisco, U.S.A, Peter MacCallum Cancer Centre, East Melbourne, Australia,Melbourne, East Centre, Cancer MacCallum Peter Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Australia, setebos s453 is a zebrafish mutant that was identified in the Liver the in identified was that mutant zebrafish a is embryos, the mRNA level of the N-terminally truncated isoform of p53, of isoform truncated N-terminally the of mRNAlevel the embryos, 1 , 4 , D. Y. R.Stainier H. Verkade is hypoplastic compared to wildtype embryos. The liver, pancreas and pancreas liver, The embryos. wildtype to compared hypoplastic is setebos 1,2 , A.J. Trotter is a premature stop codon in codon stop premature a is setebos and genes,p21 2 andJ.K.Heath 1 , Y. Rifat are also elevated. We are using Weare elevated. also are cyclinG1 embryos. This disruption culminates in culminates disruption This embryos. setebos 1 1 , A.Y.N. Ng 1 , E. Ober 4 Walter and Eliza Hall Institute of Institute Hall Eliza and Walter plus screen designed to identify to designed screen nucleolar protein 8 protein nucleolar 146 2 , H.A. Field 3 Research Division, 2 , R.D. Hannan 2 Department Department Δ113p53,is ), a (nol8), setebos 217 3 ,

Posters Posters agents forgeneticdisease. therapeutic potential as morpholinos of use the exemplifies This site. original the of forcinguse be ameliorated in the can volume ear in collapse The disorders. endolymph of study the for model a as zebrafish the in the collapse Endolymph products. truncated of generation the predicting and region, coding the in in lesions 218 gene including cotransporter sodium-potassium-chloride downregulated, the is production endolymph in role a play to thought genes several of 6 beyond However,thrive expression normal. apparently to is vesicle otic fail the of larvae Patterning fertilisation. post mutant days lethal: homozygous are lte of down. alleles upside available swimming two bladder.Mutant and The theswim circling by of function vestibular abnormal of an over-inflation signs with display the larvae together of ear, the in distension fluid endolymphatic or collapse either in ears ( Inmammals, result ear. zebrafish can The loss. hearing concomitant the inner and cochlea, the in homeostasis compartment endolymphatic inside endolymph present to fluid disruptions extracellular specialised the is Biomedical Endolymph of Department and Genetics Biomedical and Science, University ofSheffield, Developmental UK for Centre MRC T. T. Whitfield andL. Abbas the swim bladderinthezebrafish larva of volume and vesicle otic the in endolymph of regulation the for required is Nkcc1/Slc12a2 channel subunit genes. We have cloned the cloned have We genes. subunit channel 147 ) mutant shows a collapse of the otic vesicle in the larva, apparently due to a loss of loss a to due apparently larva, the in vesicle otic the of collapse a shows mutant lte) lte/nkcc1 mutant shows distinct parallels to that seen in mouse . Each allele has a point mutation that disrupts splicing, leading to frame shifts frame to leading splicing, disrupts that mutation point a has allele Each nkcc1. to27d allele by injection of a morpholino to block splicing at the new site, lte mutations and show that they correspond to correspond they that show and mutations nkcc1 ) and several and (slc12a2) Nkcc1 Organogenesis mutants, validating Na + /K + -ATPase little We collaboratingwiththelaboratoriesof are William HarrisandBrianLink. and howtheyfunctionelucidatingthemolecularpathwaysthatmediatefusion. are exploring the identity of the POM cells required for choroid fissure fusion, determining when between these cells and the retina to cells coordinate mesenchymal the periocular timing these of (POM)established,closure.is of notknowdohowthey wefunction. In our ongoing importance researchWe we hypothesize the thattheresignaling is Although closure. fissure choroid in role critical a play retina the outside cells mesenchymal that indicate others and us by studies Recent and cell movementsgeneticmechanismsthe thatabout underlieknown is choroidsevere nothing fissurecause virtually closure.fact, can this that Despite impairment. pathogeneses visual ocular common of family a colobomas, ocular in results close to fissure choroid the of Failure eye. developing the leave axons retinal and enter vessels The choroid fissure is a transient opening on the ventral side of the optic cup through which blood Anatomy, UCL, London, UK G. GestriandS.Wilson Cellular andgeneticinteractions underlyingchoroid fissureformationandclosure Organogenesis 148 219

Posters Posters 1 220 2 F. Langellotto Soul-2, ahemebindingprotein-codinggeneinkidneydevelopment University of Naples Federico II,Italy;Federico Naples of University Nitric Oxide(NO)signaling,whichisrequiredforproperHO-1activity. with and heme, of degradation the in enzyme rate-limiting a and kidney the in uptake sodium phenotypic aspects, including a mechanistic link with heme oxygenase 1 (HO-1), a regulator of additional Wecharacterizing markers. further differentiation are and regionalization of patterns and histology by shown as tubules, distal the of of Kidney development. duct pronephric during differentiation and patterning in role dual a support knockdown morpholino-driven by results Preliminary (hpf). fertilization post hours 96 until stage somites eighteen from ducts pronephric on attention our Soul zebrafish four Weidentified death. cell necrotic promote or products, iron by intoxication cell against buffer cytosolic as act may proteins SOUL heme. and porphyrins for affinity high with iron abnormal differentiation, studying kidney the SOUL/p22HBP family of During heme binding proteins, fragmentary.generic tetrapyrrole-binding factors still metabolism contributes to the development of is anemia in chronic kidney disease development patients. Weduring heme genetics are iron embryonic in of knowledge incorporated our enzymes, mainly and proteins being many in homeostasis, cofactor as body acting and for element crucial a is iron Besides of Palermo, Italy e. f ilgcl ucin, tutrs n Tcnlg, aut o Vtrnr Medicine, Veterinary of Faculty Technology, and Structures Functions, Biological of Dep. Italy; Naples,Dohrn, Anton Zoologica Biology, Stazione Developmental and Cellular of Lab. 149 orthologs, as candidate factors for heme biosynthesis and iron homeostasis. We focused We homeostasis. iron and biosynthesis heme for factors candidate as orthologs, 1,2, , A.E. Fortunato , a gene that is expressed in collective ducts and distal segment of the of segment distal and ducts collective in expressed is that gene a soul2, 1,3, , L.Castaldo 3 Dep. of Cellular and Developmental Biology,Developmental and UniversityCellular Dep. of 2 , P. Sordino imaging, as well as altered expression altered as well as imaging, vivo in morphant embryos displays thickening displays embryos morphant soul2 1 Organogenesis differentiation, including the activity of transcription factors such as thyroid in involved programme genetic The steps. successive few into subdivided is formation floor,pharyngeal its the and of midline the at tissue, endodermal the from develops thyroid the levels: molecular and ontogenetic both at mammals to comparable is fish in formation Thyroid basis ofCHisknownonlyinfewcases. genetic The disease. endocrine inherited frequent most the (CH), hypothyroidism congenital of responsible be might mechanisms these of alteration mammals, In hormones. specific produce into follicular structures and functional differentiation, leads to the formation of an organ able to process that, through the combination of migration of thyroid precursor cells, their organization in vertebrate and invertebrate organisms. Thyroid organogenesis is a The very thyroid complex is developmental an endocrine gland that produces the thyroid hormones involved in many functions and Cellular andMolecularPathology, University ofNaplesFederico II, Italy 2 I. Porreca Uncovering newgenesinvolved inzebrafish thyroid development Organogenesis 1 characterized by WMIHC (Tg, T4), of WMISH (hhex, bases cellular be will and embryos knock-down whereas approaches, loss-of-function by addressed genetic be will genes the on thyroid-expressed of role the hypotheses frame, this In follicles. new thyroid of differentiation and provide specification to aim the with orthologues, zebrafish of profiles transcriptional the of characterization the in advance will we future the In structure. thyroid of conservation the in processes anti-apoptotic of implication hypothetical a to support and is expressed in thyroid from 30hpf. Interestingly its expression disappears early in both gene this that factor. found anti-apoptotic We an is primordium thyroid zebrafish in expressed the possibility of contamination from these tissues during the LMC procedure. One of the genes raising primordium gland the surrounding regions the in expression show genes some addition, 20 genes, finding new factors that are expressed in the developing thyroid gland of zebrafish. In did of genes patterns expression the mouse analyzed have matches. we orthologous 18 date, zebrafish To show not while counterparts, mouse 56 of orthologs/paralogs 89 found 74 murine we of genes, homologs the for genome zebrafish the Surfing genes. these of counterparts fish thyroid. We are now using the zebrafish as a simple model to perform functional analysis on the gland organogenesis. This approach highlighted a list of genes that are enriched in mouse E10.5 oligonucleotide Microdissection)-based Capture (Laser microarray analysis was LMC performed in mouse an to identify genes differentially disease, expressed during the CH in development, therefore thyroid and normal in implicated machinery genetic the on insights further Togain conserved withrespecttoexpressionpatternsandfunctions. type, mutant(noisthmus,handsoff)andtransgenic(Tg(nk2.1a :YFP) backgrounds. Biogem,Italy; Institute Naples,of Genetic Dohrn,Research Anton “Gaetano Salvatore”,Zoologica Biology, Stazione Developmental Ariano Irpino,and Italy; Cellular of Lab. morphant embryos when the thyroid primordium is still present. These results lend results These present. still is primordium thyroid the when embryos morphant Hhex 1,2 , H.Fagman 2 , M.DeFelice 1,2 , P. Sordino nk2.1a, 1 , R.DiLauro pax8, pax2.1, 3 150 Nkx2.1a, tg) and TUNEL in wild- Pax8 and 3 Dep. of Biology Nkx2.1a Hhex, is 221

Posters Posters 1 as astrongcandidateforcartilagemaintenancediseasesand craniofacialbirthdefects. prospective on the Sec24D cargo specificity during development. Our findings establish Sec24D Sec24D is indispensable for chondrocyte maturation. This study presents the first developmental indicating that Sec24C can compensate for loss of Sec24D at initial stages of chondrogenesis, but development in double Sec24C/Sec24D-deficient animals is arresteddysmorphology.craniofacial in However,result earliernot craniofacial does Sec24D, of paralog than close BiP and activation of the ER stress response machinery. We found that knockdown of Sec24C, a of upregulation to leads ER the in proteins of backlog The functional. appear staining ceramide 2 222 E. W. Knapik Sec24d transports extracellular matrixproteinsduringzebrafish skeletalmorphogenesis Furthermore, Sec24D-deficient cells accumulate proteins in the distended ER, although a ER, thedistended in proteins subset of ER compartments and Golgi complexes visualized by electron microscopy and NBD accumulate cells Sec24D-deficient Furthermore, fashion. Sec24D-independent in transported are molecules adhesion cell and b1-integrin since receptors bound membrane of lack a not and deficits, matrix from stems development cartilage and small dysmorphology.craniofacial severe and cartilages malformed in We in arrest the that demonstrate resulting thus intercalate, and shape change to normally. fail proceeds chondroblasts Consequently, condensations pre-chondrogenic and primordia for craniofacial critical of proteins matrix migration extracellular of transport whereas chondrocytes, mature by matrix mutation zebrafish cloned cargo are the positionaly We development have complexes. COPII the of component integral an duringvertebrate however,is Sec24D adaptor Cargo proteins, adaptors transport model ofindividual largely unknown. using role approaches the and biology specificity studied cellular been have and mechanisms binding biochemical Cargo COPII. as such coated by carriers conducted primarily vesicular is Golgi to (ER) reticulum endoplasmic from transport Protein Biological Sciences, Vanderbilt University, Nashville, USA, iiin f eei Mdcn, adrit nvriy eia Cne, ahil, USA; Nashville, Center, Medical University Vanderbilt Germany; Freiburg, of Medicine, University I, Biology Biology, Institute Genetic Developmental of Division 151 1 , S.Sarmah 1 , A. Barrallo-Gimeno and shown that Sec24D is essential for secretion of cartilage of secretion for essential is Sec24D that shown and bulldog 2 , J.Topczewski 3 , L.Solnica-Krezel Organogenesis bulldog/sec24d, 3 eatet of Department 3 by vitro in 6 - Brown University, R. Creton and behavioral changes Modulators ofcalciumsignalinginducedevelopmental brain defects Neurobiology I:Patterning nomto o te eeomna mcaim, idw o sniiiy ad dose-response and sensitivity, of windows mechanisms, relationship is essential for risk assessment of pharmaceuticals that modulate calcium signaling. developmental the on information include convergence extension during gastrulation and the formation of Kupffer’s vesicle. laterality in the Further brain and asymmetry in of development behavior.the affect time The this underlying during Exposures calcium-sensitive mechanisms gastrulation. during signaling calcium of affect signaling calcium of modulators how brain developmentandbehavior. examine We foundthatzebrafishembryosaresensitivetomodulation to system model a as zebrafish use We developmental times, and sensitive the potentially the pleiotropic on effects information on brain of development lack and indirectly,a behavior.or directly either signaling calcium affect can that pharmaceuticals of number large the of because evaluate to difficult is development brain manipulations calcium subtle that during embryonic development can induce specific brain defects. The potential risk for human shown have systems model animal in studies as troubling, concentrations of pharmaceuticals in the environment and drinking water. These exposures are embryos Human trace by or pregnancy development. early during medicine of use embryonic maternal by exposed be during inadvertently may exposure for risk increased an comes use, pressure, heart arrhythmias, blood high angina treating pectoris, for used and successfully are migraine. signaling calcium modulate that However,Pharmaceuticals along with their widespread 1 , J. A. Kreiling 1 MCB Department, 1 , andR.M.Colwill 2 Psychology Department, Providence, RhodeIsland, USA 2 152 223

Posters Posters E.M. Mignot le aon ad le cnoiain in consolidation sleep and amount sleep and nocturnalmammals,theroleofthiscircuit inotherspecieswillneedfurtherinvestigation. diurnal both in described HCRT-Pinealbeen similar have As system. projections melatonin the these two major sleep regulator pathways and suggests a HCRT sleep consolidation role through of phenotype fragmentation sleep the of presence abrogates which melatonin, The to hypersensitivity striking compounds. the and projections hypnotic gland pineal HCRT other using genotypes between found was difference characterized further we melatonin response in dark, the in released is and zebrafish diurnal in hormone promoting to the pineal gland, a brain structure expressing parallel with established mammalian HCRT neuroanatomy. We also found significant projections expressing stably zebrafish HCRTin circuitentire Visualizationthe of decarboxylase acid glutamic in mammals, zebrafish HCRT neurons express vesicular glutamate transporter sleep of like species. that, this found HCRT in We characterized neurons mediators we zebrafish, in consolidation potential investigate zebrafish. To and in consolidation, conserved sleep is phenotype also later but the wake that only not regulates HCRT that suggests This mutant (hcrtr-/-). receptor HCRT zebrafish in recapitulated phenotype understood less a fragmentation, deficiency HCRT and consolidate to however,wake fragmented sleep to display is addition also In narcolepticmammals wakefulness. wake-promoting system HCRT the of function strongly major a that is indicates This narcolepsy. causes (HCRT) hypocretin/orexin mammals, In Institute ofGenetics, Mishima, Japan 1 Stanford Stanford University, Palo Alto, USA; 2 224 Appelbaum L.A. Hypocretin interacts withmelatonininregulatingsleepzebrafish eatet f scity n Bhvoa Sine, tnod nvriy Pl At, USA; Alto, Palo University, Stanford USA; Sciences,Alto, Palo University, Physiology, Stanford Behavioral Cellular and Molecular and Psychiatry of Department 153 1 , P.M. Mourrain 1 G.W. WangG.W. , hcrtr-/- 1 larvae and adults. Interestingly, a low dose of melatonin increases niaig osrain f h guaaegc phenotype. glutamatergic the of conservation indicating gad67 2 , G.M. Maro G.M. , 4 Division of Molecular and Developmental Biology, National fish demonstrate a specific interaction between interaction specific a demonstrate fish hcrtr-/- hcrtr-/- 3 , W.M. Marin W.M. , hcrtr mRNA. As pineal melatonin is a major sleep fish u nt n hi wltp sbig. No siblings. wildtype their in not but 1 , T.Y., Yokogawa Neurobiology I:Patterning revealed clear revealed hcrt:EGFP 3 ilgcl Sciences, Biological 1 , K.K. Kawakami K.K. , vglut2 but not the 4 , Neurology and Pediatrics, University of Utah, USA;Utah, of University Pediatrics, and Neurology 1 J. Schweitzer diencephalospinal axons Netrin-DCC, Robo-Slit and HSPGs coordinate lateral positioning of longitudinal dopaminergic Neurobiology I:Patterning long-range cues from the midline act in a concerted fashion to define lateral positions of DA of positions lateral define to longitudinalaxontracts. fashion andattractive concerted a in act ofrepulsive midline the from cues integrations long-range Simultaneous pathfinding. diencephalospinal DA for the complexity of positioning far-projecting longitudinal axons, and allow us to provide a model mechanisms. guidance diencephalospinal DA of modulators essential as proteglycans sulfate heparan identified Wefurther tract. diencephalospinal DA the signaling. of positions DCC/Netrin1 lateral specify to by attraction mediated midline Netrin counteracts repulsion the based Slit towards Thus attracted are axons DA function, Robo2/Slit repulsive of absence the In midline. the from away axons DA longitudinal keeps signaling Robo2/ Slit Repulsive cues. derived midline of signaling range long integrating by navigate axons mutant Using activity. DAlongitudinal locomotor that demonstrate we experiments function of modulating ventralloss and gain- as well as in analysis the involved in be located to neurons thought DA is by and established diencephalon is tract This tract. diencephalospinal (DA) are largely unknown. Here we study axon guidance mechanisms mechanisms specifying of the the formation of longitudinal longitudinal axon dopaminergic tracts in the vertebrate nervous system characterized, well been have formation pathway commissural driving determinants molecular Longitudinal axons provide connectivity between remote areas of the nervous system. While the Switzerland Neurobiology and Anatomy, University of Utah, Salt Lake City,USA;Lake Salt Utah, Anatomy,of and University Neurobiology Developmental Biology, Institute Biology 1 University of Freiburg, Germany;Freiburg, of University 1 Biology Biology,Institute Developmental 1 , E.Kastenhuber 1,4 , U.Kern 1 , J.Bonkowsky 4 Institute of Zoology, University of Zurich, Zoology,of of University Institute 2,3 , C-B.Chien Our findings provide insight into insight provide findings Our 154 3 2 Department of Pediatric of Department andW. Driever 2 Department of Department 1 225

Posters Posters 1 leads totheestablishmentofdiscreteneuronaldomainsinthalamus. neurons. inhibitory GABAergic, Thus, of the presence production or absence Ascl1-mediated of a to single upstream leads regulator of proneural Her6 gene expression, of Her6, maintenance forcedwhereas neurons, glutamatergic,excitatory of genesis Neurogenin1-mediated premature of neurogenesis but also, as important, the subsequent identity of neurons. Loss of Her6 leads to progression and onset the only not determines Neurogenetic Her6 that show we Here Ascl. and Neurogenin the as to as such genes, proneural regulating by neurogenesis control to shown been has family Hes/Her referred manner gene bHLH the by regulation its and thalamus zebrafish the in regulatedgradient neurogenetic spatially a Gradient, the underlying mechanism of which is unknown. Here we describe the existence of a in proceeds thalamus mammalian the neurogenesis In regulated. strictly are neurogenesis of progression and onset The Umeå Center forMolecularMedicine, UmeåUniversity, Sweden. Developmental Neurobiology,Developmental Hunt’sNew House, Guy’s Campus, King’s CollegeLondon, UK; 226 S. Scholpp Her6 regulatestheNeurogeneticGradient andNeuronalIdentityinthe Thalamus Institute of ToxicologyInstitute Genetics,and Forschungszentrum Karlsruhe, Germany; 155 1 , A. Delogu 2 , J.Gilthorpe 2,3 , D.Peukert 1 and A. Lumsden Neurobiology I:Patterning 4 2 MRC CentreMRC for . The her6. 3

A modeloftheunderlyingmolecularmechanismexplainingphenotypewillbepresented. trigeminal the of ganglion inzebrafishembryoslackingzfGrx2. loss as well as CNS the in tracts axon and neurons of reduction significant a Sweden; 1 Knock-Down of a Glutaredoxin (Zebrafish Glutaredoxin 2) Leads to Impaired Neuron toImpaired 2) Leads L. Braeutigam Glutaredoxin (Zebrafish Glutaredoxin Development a of Knock-Down Neurobiology I:Patterning hrceie b aotss n h CS uig dsic dvlpetl ie idw from window, time by TUNEL staining confined and were developmental cells apoptotic hpf, distinct 24 At (hpf). a fertilization post hours during 48 is to CNS midsomitogenesis It the phenotype. in neurodegeneration apoptosis a by in characterized results zfGrx2 CNS. of the knock-down in neurons induced in differentiated Morpholino to whereas confined mainly distributed, was protein areubiquitously zfGrx2 situation adult the transcripts zfGrx2 embryogenesis zebrafish During in coordination as [FeS] cluster implies gelfiltration [2Fe2S] protein monomers. an homologue, of human coordination its to the contrast In indicated well. zfGrx2 recombinant of spectra UV/Vis a homologueofhumanGrx2. Here, we report the first between twomonomersandprotectscellsagainstoxidativestressinducedapoptosis. of regulators major the intracellular and redox potential. The human mitochondrial hGrx2 coordinates proteins a [2Fe2S] cluster of family Thioredox the of members are (Grx) Glutaredoxins Hogskola,Sodertorns Stockholm, Sweden hindbrain and spinal cord. spinal and hindbrain Phillips Universoty Marburg, Germany; Biosci.Nut.,Institutet,and Karolinska Stockholm, Sweden; 4 Medical Biochemistry, Karolinska Instituet, Stockholm, Sweden; 1 , J.G.Berthet in vivo Caspase-3 activity measurements predominantly to the forebrain, in vitro and 2 Acetylated tubulin, Acetylated , I.Soell in vivo characterization of a dithiol Grx in zebrafish (zfGrx2), 3 , A. Holmgren 3 School of Life Sciences, Sodertorns Hogskola, Stockholm, and zn5 4 , C.H.Lillig immunohistochemistry revealed immunohistochemistry HuC 2 Clinical Cytobiol. and Cytopathol.,Cytobiol.and Clinical 2 , G.Hauptmann 156 5 School of Life Sciences, 5 , C.Berndt 227 4

Posters Posters 228 spawning groupsandaltersreproductivecapabilityoutcome. in interactions social and behaviour on impacts subsequently life early during EE2 to exposure to the 10ng/L EE2, compared with those exposed to 3ng/L EE2. These results strongly suggest that the in colonies dominant in were most cases. they Unexposed and females spawned females, fewer exposed eggs when with courted compared by success males exposed reproductive higher significantly had females unexposed the females, unexposed or exposed courting in preference was modified significantly byfemales EE2the exposureof duringsuccess Reproductive earlyfemale. life.1 Whilstand unexposedmales life males2 showedwith no conditions early for pair remaining during concentration EE2 highest the between courtship preventing of the point the to adults, as aggression of to level higher a showed exposed males and EE2, and of behaviourconcentration exposure reproductive that subsequent found on We consequences exposure increasing with reduced significantly were attempts courtship Male success. lastingbreeding female. long one had for life competing early males during two or male, one with females two either with colonies in adults as success breeding and behaviour courtship their observed water,subsequently aquarium and standard in months 7 for depurate to them allowed dpf), 60 of ng/L 10 and 3 to – (20 differentiation sexual of period the zebrafish encompassing life, early during (EE2) ethinyloestradiol exposed we where study recent a on report we Here outcome. This breeding the and microsatellites). interactions these on DNAEDCs of effects for probe of and success reproductive and use the (through colony the experimental in approach allows us to establish dynamic relationships between dominance, spawning effort breeding the on and EDCs affects reproductive behaviour and the impacts this has courtship on breeding and success for reproductive individuals alter Using to behaviours. and ducts, reproductive of malformation and inter-sex as such conditions inducing fish, exposed of physiology the affect to shown been have chemicals These waters. receiving their and effluents works treatment wastewater in found commonly are Endocrine disrupting chemicals (EDCs), including environmental oestrogens and anti-androgens, School ofBiosciences, Altering University ofExeter, UnitedKingdom Behaviour Reproductive Subsequent M.K. Sýffker, T.S. Coe, A.L. Filby, C.R.Tyler on Impacts Estrogen Breeding OutcomeinZebrafish (Daniorerio) Colonies to Exposure Early-life 157 (zebrafish) in the laboratory, we are assessing how exposure to exposure how assessing are laboratory, we the in (zebrafish) rerio Danio Neurobiology I:Patterning 77. fate. Biol. Dev cell 295:263- brain: and dynamics, origin,zebrafish migration proliferation adult Grandel H, Kaslin J, Ganz J, Wenzel I, Brand M. (2006).mammalian vertebrate brain. Philos TransNeural stem RSocLond BBiolSci.cells 29;363(1489):101-122.and neurogenesis in the Kaslin press J, J,Ganz M.Brand (2008). in Proliferation, Neuroscience,non- the in regeneration and neurogenesis of Journal niche, cell stem novel a of maintenance and initiation cerebellum: Kaslin J, Ganz J, Geffarth M, Grandel H, inthe Hans S, Brand cellregulation M. (2009). stem Stem cells neural in the adult adult zebrafish of studying context oftheiradultanatomicalsettingin-vivo. importance the highlight findings considered. These zone proliferation telencephalic individual the on depending signaling, Bmp and Fgf for requirements activity. distinct niche Surprisingly,find neurogenic we adult regulating in function approaches, we demonstrate important functions of the Fgf and Bmp signaling pathways the niches in zebrafish are similar to mouse in their molecularcells and composition.the migration characteristics transit-amplifying of the Usingneuroblasts in of the ventral gaintelencephalon. However,arrangement and spatial lossthe of in example for differences display telencephalon comparingthe by We Furthermore, gliacharacteristics. domains. neurogenic niches between mouse and radial zebrafish we reveal that the typical niches in glia the adult zebrafish the display in not pool and progenitor does the of zebrafish part arrangements major a view, common cellular the to contrast in characteristics, that, demonstrate proliferation distinct telencephalon ventral and dorsal display the in niches different four the that show we telencephalon By analyzing the cellular and molecular composition of neurogenic niches in the adult zebrafish types andsignalsregulateneurogenesisinthedifferentneurogenicniches? cell which organized,and niches neurogenic different the are How questions: key several raise observations These 2009). al., et Kaslin 2006, al., et (Grandel brain zebrafish adult the of axis central the in process that distinct neurogenic niches are lifelong present in different brain parts along widespread the whole rostral-caudal a is We2008). shown al., previously et have (Kaslin vertebrates non-mammalian of neurogenesis system nervous adult mammals, to contrast In Dresden, Germany Biotechnology Center and Center for Regenerative Therapies Dresden, University of Technology characteristics, proliferation J. Ganz,Kaslin,H.Grandel,D.Freudenreich,S.Hochmann,M.Brand telencephalon: zebrafish adult the cellular compositionandregulationby fgfandbmpsignaling in niches neurogenic The Neurobiology I:Patterning 158 229

Posters Posters 230 suggest thatNetrinsignalingisrequiredforVaP development. beyond their typical time of death. Collectively, the dcc-MO injected embryos, presumptive VaP axons extended beyond the MPs and VaPs survived knockdown revealed that, like Ntn1a, Dcc is required for development of VaP but not of CaP.at In stop to axons VaP cause MPs. to the Ntn1a of ability the mediates Dcc whether examined also We axons fromextendingintoventralmuscle. VaPpreventing signal VaPs.MP-derived wildtype the than be longer may survived Ntn1a Thus, CaP.In of not but VaP VaPof motoneurons presumptive the embryos, and injected MPs MO the beyond extended axons development for required is Ntn1a that revealed VaPs, and CaPs presumptive dye-labeled of observations interval by followed (MOs), morpholinos using Zebrafish survival. cell and Ntn1a down VaPKnocking and CaP by cones. contact growth guidance of time the at MPs in axon expressed in roles with ligand secreted a is Netrin that mediatestheabilityofNtn1atocauseVaP axonstostopattheMPs. VaP VaPfor receptor WeNetrin a carcinomadevelopment. as colorectal (Dcc) in Deleted identify also required are MPs the and for necessary signal CaPmolecular MP-derived an as (Ntn1a) 1a Netrin identify we Here formation. both that showed work dorsal Previous divide muscle. that fibers ventral short muscle and slow extend identified and (MPs), pioneers hemisegments muscle spinal the at the stop of that axons half about in present are VaPs muscle. ventral typically dies. CaPs are present in all spinal hemisegments and extend long axons that innervate VaP,CaPand later but pair equivalence VaP an form initially motoneurons, zebrafish identified Institute ofNeuroscience, University ofOregon, Eugene, USA L.A. Hale,J.S.Eisen Netrin SignalingisRequiredforDevelopment ofanIdentifiedZebrafish Motoneuron 159 s xrse i CP n VP hogot xn ugot. Omdae Dcc MO-mediated outgrowth. axon throughout VaP and CaP in expressed is dcc dcc-MO and Neurobiology I:Patterning ntn1a-MO experimental results is ntn1a ntn1a- 1 V.T. Cunliffe progenitor fate neural of control epigenetic in 1 deacetylase histone of action of mechanism the Dissecting Neurobiology I:Patterning University ofSheffield, UK, and of ventricular zone neural progenitors as well as other genes that influence the behaviour of behaviour the influence that behaviour genes the differentiating, post-mitoticcellsinthemantleregion. other as control well that as genes progenitors neural regulate zone may ventricular of Hdac1 that suggest Taken also complexes. results silencing our transcriptional together, distinct multiple of component a as development in the developing CNS. The results of these experiments indicate that Hdac1 may promote neural embryos lacking the functions of other chromatin of regulators phenotypes that the are of likely analysis to comparative a interact out with carry Hdac1 to markers as genes genes activity.Hdac1 targetsof downstream as implicated are Hdac1-regulated that We Hdac1-regulated the used also of functions the analysing and chromatin, in genes target to Hdac1 of binding the investigating now Weare CNS. developing the in Hdac1 of targets direct potential the of analysis expression dissect further the mechanism of action of ToHdac1 genes. target of in vicinity neurogenesis, the in we acetylation performed histone limiting a by genome-wide genes repress may Hdac1 that acetylation during embryogenesis and specifically promotes histone methylation, which suggests its known in vitro deacetylase activity, we find that Hdac1 regulates steady-state levels of histone precise molecular mechanisms of neural Hdac1 action in neurogenesis of remain unclear. commitment Consistent with promoting in Hdac1 progenitors and for differentiation of post-mitotic role neurons via expression of regulatory proneural genes, but a the identify observations These a mechanism that antagonises the Notch pathway-mediated maintenance of neural progenitors. results further show that Hdac1 promotes expression of proneural genes throughout the CNS by 1 (Hdac1) in regulating the pace and pattern of both neurogenesis and oligodendrogenesis. Our stringent epigenetic control. Our studies have revealed critical functions for Histone Deacetylase The transformation of neural progenitors into differentiated neurons and myelinating glia is under MRC Centre for Developmental & Biomedical Genetics and Department of Biomedical Science, 1 , M.R.Harrison mutant phenotype and identified several Hdac1-regulated, several identified and phenotype mutant hdac1 1 , A.S. Georgiou 2 Institute ofBiology, Leiden University, The Netherlands 1 , E.G.Lightman 1 , A.J. Buckle 160 1 , H.P. Spaink mutants and mutants hdac1 2

231

Posters Posters 232 of genesregulatingpatterningtheneuralretinaandlens. inhibitor. Taken together, these data indicate that Gdf6a functions at the top of in the lens Bmp development. These hierarchy defects are enhanced by addition of Dorsomorphin, a selective Bmp phosphorylation in the lens and for the expression of lens-specific genes, leading to mild defects Finally, we demonstrate a role for Gdf6a in non-neural ocular tissues. Gdf6a is required for Smad affects dorsal,butnotventralgeneexpression. retinal dorsal induce ventral markers due to Gdf6a inhibition is independent of can Tbx5 function, as knockdown of Tbx5 Gdf6a of expansion the that Wedemonstrate patterning. also dependent Gdf6a for that patterning, required not is retinal but dorsal drive show to sufficient is Bmp4 also Thus, Bmp4. of absence but the in patterning defects, patterning dependent rescue can Gdf6a Bmp4 of over-expressiondata as provide Gdf6a, Weto related development. functionally eye is of Bmp4 that stages indicating early at markers dorsal of expression induce to sufficient is over-expressionGdf6a through that and analysis, morphants, Gdf6a are in initiated not fate cell dorsal of markers earliest the that show clearly We domain. dorsal the into retina of loss phosphorylation, 1/5/8 Smad abolishes function Gdf6a of knockdown morpholino zebrafish, In Gdf6a, isessentialforthisprocess. it isprofoundly that on dorsal retinal specification. Rather, our effect andfound research discernable no implicates a produces related Bmp4 Bmp zebrafish signalingof molecule, Knockdown model. zebrafish chick the from different in specification retinal dorsal-ventral have investigated We experiments. to over-expression and leadingknockdown both by axis, evidenced as nasal-temporal identity, dorsal and dorsal-ventral the along carefully orchestrated retinotectal axon mapping. In chicken patterned embryos, Bmp4 signaling specifies is retina neural The ofBiological Sciences,Department University of Alberta, Edmonton, Canada C. R.French, T. Erickson,D.V. French,D.B.Pilgrim, A. J.Waskiewicz Gdf6a isrequiredforinitiationofdorsal-ventral retinalpatterningandlensdevelopment 161 expression in the dorsal retina, dorsal the in expression tbx5 and expansion of expansion and Neurobiology I:Patterning expression from the ventral the from expression vax2 for thezebrafishproteininOSdevelopment. axon OSN block fully not does elongation or depletion OB differentiation, as anosmin-1a it is the zebrafish case in KS that patients, suggesting a showed distinct requirement also results our that full differentiation OB glomeruli relies on their proper innervation by OSN axons. However, growth and guidance of OSN axons and OB neuron differentiation. In addition, our proper data for suggest protein the of role conserved evolutionarily an revealed vertebrates in GABAergic development of number OS the in anosmin-1 for inrequirement the of investigation decrease first This neurons. severe OB dopaminergic and a induces and glomeruli OB of that differentiation showed also We axons OBs. within OSN targeting of terminal fasciculation their the and compromises depletion anosmin-1a that found We anlagen. onward, post-fertilization hour 22 from (OSNs) neurons of use made olfactory system (OS) development in vertebrates. We detected anosmin-1a in olfactory sensory we Here, differentiation. (OB) bulb cell zebrafish (Danio ) rerio embryos olfactory impairs as model to investigate that and disorder elongation, genetic axon neurological migration, human a (KS), syndrome Kallmann underlies activities, -branching and axon-guidance and outgrowth, neurite adhesion, The salpêtrière, Paris, France I.C.M),(CR Pitié- épinière Moelle la Hôpital de Cerveauet du l’Institut CentreRecherchede de olfactory of C. targeting Yanicostas, E.Herbomel, A. Dipietromaria,andN.Soussi-Yanicostas terminal and fasciculation sensory neuronaxonsinzebrafish embryos in Anosmin-1a for requirement Essential Neurobiology I:Patterning ee noe aomn1 a etaellr arx rti, hc mdae cell mediates which protein, matrix extracellular an anosmin-1, encodes gene KAL-1 in vivo the requirement for this protein in prior their pioneer axons reach OB reach axons pioneer their prior i.e. nciain mar the impairs inactivation kal1a 162 . This gene This . vitro in 233

Posters Posters 1 signalling pathwaysintheepithamus. address this issue, we are currently working on resolving the interactions between Nodal Toand Fgf cells. parapineal and/or habenular in activity pathway Fgf modulating by and/or precursors could do so through promoting a left/right asymmetry in the number of it activity; Fgf8 of levels biasing by laterality parapineal impose could signalling Nodal whether determine to important is it Therefore, 2009). al., et (Regan migration of direction the bias can Nodal-dependent on dependent early is migration parapineal which that shown in model a propose we asymmetry studies, in habenular neurogenesis previous would bias and parapineal migration towards data the left. We our have From than indirectinglateralityonly(Roussigneetal.,2009). asymmetry an promoting in signalling Nodal for requirement direct a of evidence a revealing symmetric provide neurogenesis results our Thus, neurogenesis. biasing asymmetrically habenular in pathway this for requirement renders signalling Removing Nodal parapineal. in the bias on left-right dependent the not is neurogenesis in difference left-right temporal this phenotypes, asymmetry habenular other to contrast in and, right the in than habenula left the in earlier present are neurons that recently shown Wehave signalling. Nodal unilateral by requires the presence of a second asymmetric structure, (L/R) the parapineal, left-right whose laterality display is habenulae biased the asymmetries zebrafish, in In midbrain. gene ventral expression the and to axonal forebrain the projections. from The elaboration of habenular asymmetries input relay that epithalamus the in nuclei bilateral conserved evolutionarily are habenulae The UCL, London, UK; 234 M. Roussigne Nodal signallingimposesleft-rightasymmetryuponneurogenesisinthehabenularnuclei CellDevBiol,and UCL, London, UK; 163 1 , I.H.Bianco 4 CBD, Université P.Sabatier, Toulouse, France 2 , S.W. Wilson 2 Harvard University, Harvard Cambridge, USA; 3 , P. Blader and L/R differences in the levels of Fgf8 of levels the in differences L/R and fgf8 4 Neurobiology I:Patterning fgf8 expressing habenular 3 CellDevBio,and rather se per one ofmonitoringsystemthatensuresretinalneurogenesisinzebrafish. Taken mutants. these together, is regulation apoptotic p53-mediated the that suggest data these apoptotic zebrafish other mutants: in p53 on depends apoptosis retinal whether examined we mutant, the to addition In retina. zebrafish the in apoptosis and differentiation cell between choice fate ATM/Chk2/p53the cell on the pathway.depends influences mutant p53 Wethat conclude subsequently activates a p53-dependent apoptosis. We found that neuronal apoptosis in the activate Ataxia telangiectasia mutated (ATM). ATM activates Checkpoint kinase 2 (Chk2), which may which DNAcheckpoint, the replication triggers Prim1 RNAby that synthesis reported was the in Y. Yoshimura Mechanisms regulatingneuronalapoptosisinthedeveloping zebrafish retina Neurobiology I:Patterning 1 ifrnito. mses mtto ocre i te ml sbnt f N piae (Prim1) primase DNA of subunit small the the in in occurred mutation missense A differentiation. Herewereported retina. in the developing them, of one apoptosis extensive show that mutants zebrafish five identified we issue, this Toelucidate development. during regulated is pathway apoptotic the how unclarified However,remains tissues. it developing the in observed often is Apoptosis Okinawa, Japan eeomna nuoilg ui, IT Oiaa Japan; Okinawa, OIST, unit, neurobiology Developmental piy mutant, suggesting that this missense mutation does not affect DNA replication. It replication. DNA affect not does mutation missense this that suggesting mutant, piy , rw329 mutant. Prim1 is required for DNA replication, but cell proliferation seems normal seems proliferation cell but replication, DNA for required is Prim1 mutant. 1 , N.Hanahara , in which most retinal neurons undergo apoptosis during their during apoptosis undergo neurons retinal most which in (piy), eye pinball and rw337 1 , M. Yamaguchi . We found that retinal apoptosis also depends on p53 in p53 on depends also apoptosis retinal that found We . rw564 1 , N.Fujimori 2 , I.Masai 2 ntaie eerh nt RIKEN, Unit, Research Initiative 164 1 235 piy piy

Posters Posters 236 anti-acetylated MMP14a, of Knockdown pathway. / tectum using a specificoptic MMP14a morpholino, and subsequentdeveloping immunostainings the in present are MMPs these extensive an show expression pattern in the developing zTIMP2 brain. Cross-sections of these and zMMP14b zMMP14a, zMMP2, Especially hybridization. situ by system retinotectal the of development during points time various at embryos zebrafish of We characterized the expression of several MMPs and TIMPs, identified in zebrafish, in the brain in retinotectalpathfindingremaincompletelyunidentified. metalloproteinases) of inhibitors (tissue MMPs/TIMPs these of the mechanisms However,working and projections. nature retinotectal of development the in participate substantial MMPs also that is evidence There brain. developing the in myelination and guidance axon neurons, of Recently matrix metalloproteinases (MMPs) have been shown to regulate migration and survival visual pathway. revealed several attractive and repulsive guidance cues that regulate the proper formation of the models animal formation different in and studies loss/gain-of-function crossing Recent midline maps. neuronal guidance, topographic of axon study to system model powerful a is (RGCs), cells ganglion retinal of circuit,axons optic the The contains axons. which their and neurons of Correct wiring of neuronal circuits in the developing brain relies on the precise spatial positioning Belgium Biology,of Development Circuit RegenerationNeural Department and ResearchGroup Leuven, E. Janssens andL.Moons Developmental retinotectalaxonpathfindinginzebrafish: aroleformatrixmetalloproteinases development of the visual pathways and that axon pathfinding in the retinotectal system of system for futurestrategiesinterferingwithneuralcircuit developmentand/orregeneration. retinotectal the in pathfinding implications have might results Our axon signaling. MMP of blockade by disrupted be that can zebrafish and pathways visual the of during brain development zebrafish the in expressed are MMPs that reveal findings novel these Altogether, retinotectal axonalpathfinding. broad-spectrum in role the a play indeed with MMPs that obtained show and hypothesis were our confirm EDTA.They inhibitor results MMP Similar (p<0.05). tectum optic the of size the 165 uui, t df dy ps friiain showed fertilization) post (days 5dpf at tubulin, α Neurobiology I:Patterning in situ stainings revealed that a significant reduction in reduction significant a with the axonal marker in D. Peukert Function oftheLhxgenesinthalamicdevelopment ofthezebrafish Neurobiology I:Patterning 1 population does not express not does population and anterior ZLI border and the posterior located pretectum (Scholpp et al., 2006). We find that of pattern organism.expression the mapped model we First, a as zebrafish using by detail in process parcellation the elucidate we Here, genes homeodomain LIM the that Xenopus show and mouse from Data unknown. are nuclei the units: functional its of formation and mammalian lineage. Interestingly, the developmental process of regionalization of the thalamus area in the cortex. Although much simpler, this basic principal can be observed within the non- manner: stereotypic a in sensory organized organs is project information towards of specificflow thalamicthe nucleimammals from whichIn subsequentlyparts. with targetbrain organs asensory higher the specific connects and forebrain the in station relay major the is thalamus The and Genetics, Research Karlsruhe centre (Helmholtz-Gemeinschaft), Germany nuclei. To explore this further we will study the function of these LIM homeodomain factors, homeodomain LIM these which willgiveusagreaterinsightintothalamicdevelopmentinvertebrates. of function the study will we further this Toexplore nuclei. final the into population this of process parcellation further the during role crucial a suggesting neurons, projection thalamic the within domains overlapping partially but distinct in expressed Germany; nttt fr oiooy n Gntc, eerh ete alrh (Helmholtz-Gemeinschaft), Karlsruhe centre Research Genetics, and Toxicology for Institute mark the pool of thalamic postmitotic projection neurons, whereas the interneuron the whereas neurons, projection postmitotic thalamic of pool the mark lhx9 2 1 CentreDevelopmentalfor Neurobiology, MRC, London, UK; , A.Lumsden 2 , S.Scholpp and lhx2 3 . Furthermore, we could show that show could we Furthermore, lhx9. are expressed in the different thalamic compartments. thalamic different the in expressed are and lhx2 relative to well-known markers of the of markers well-known to relative lhx9 166 3 Institute for Institute Toxicology and lhx2 are lhx9 237 lhx2

Posters Posters key generegulatorynetworkthat,whendisrupted,causesHPE. the a of understanding our advance attenuate and forebrain developing the in to expression gene Hh–activated of modulator acts a as Zic2a Zic2a for role tissues essential novel, both a uncover data in These signaling. increasing Hh that of by effects suggesting or Hhip, signaling antagonist Hh Hh blocking the by of rescued are levels OS/retina the in and forebrain the in targetsHh of transcription and (OS) stalk optic diencephalon-derived neural The retina are also patterned prethalamus. in response to Hh signaling. the We show that of zebrafish Zic2a limits formation correct the for of repression Zic2a signaling. Hh limiting through likely domain, expression developing forebrain. developing of transcription activates signaling Hh mouse, in as zebrafish, in that demonstrate Shh and Six3 mouse, In activate each other’s Zic2. transcription, but a role for Zic2 in and this interaction has not been tested. Six3 We factors transcription and (Shh) hedgehog Sonic factor in forebrainbecause they cause HPE when mutated. the Among these are genes of identified that encode been the have secreted development growth malformation forebrain during roles essential congenital with genes Several commonhumans. most the is (HPE) Holoprosencephaly Ph.D. Training Program, University of Wisconsin, Madison, USA 238 Y. Grinblat A Novel RoleFor ZicGenesIn The Developing Forebrain 2 Departments Departments of Zoology and Anatomy, University of Wisconsin, Madison, USA and 167 1 , N.A.Sanek 2a is also activated by Hh signaling and represses and signaling Hh by activated also is zic2a 1, , A.A. Taylor 2a and pax2a 8a in the OS and retina. The effects of Zic2a depletion Zic2a of effects The retina. and OS the in fgf8a 2 , M.K.Nyholm 2 Neurobiology I:Patterning 3b outside of its of outside six3b 3b is essential is six3b 1 the Genetics 3b in the in six3b role during development as a regulator of the neuronal progenitor neuronal the of regulator a as development during role important invivo. This is the first genetic evidence that cullin box domain of an ASB-family member is functionally as well as well as (her4,her1,genes her6) target Notch of upregulation by demonstrated as activation signaling Notch in involved is cullin Asb11-bindinggenes. to these of expression correct for required is box cullin that shows mutants the in genes pathway Notch we Asb11 of domain box employed cullin Tillingits to of albeit generate a zebrafishrole complex, mutant deleting theinvestigate cullinECS To box. The expressionthe unknown. of Delta/ form remains to function box biological cullin of role the demonstrated have studies Several DeltaA, with interacting suggesting that of d-Asb11capable may not mediate is ubiquitination domain as box a SOCS specificthe componentlacking d-Asb11 of activation. the ECS complex. ubiquitination and degradation of the Notch-ligand DeltaA, positively regulating Notch signaling 1 M. SartoridaSilva* Role ofcullin-bindingdomain Asb11 inneurogenesis Neurobiology I:Patterning ubiquitination and degradation. Weidentified degradation. and for ubiquitination them targeting by proteins of half-life the regulating (elonginC–cullin–SOCS- complex ligase ECS ubiquitin an E3 of type box) part as act (ASB), proteins box SOCS and repeats Ankyrin Netherlands urct nttt, teh, h Netherlands; The Utrecht, Institute, Hubrecht where mutant Asb11mutant where vitro in 1,2 , J-M.Tee* 1 , A. Brouwers

has a reduced capacity to induce Notch reporter Hes1. reporter Notch induce to capacity reduced a has 1 , S.Diks d-asb11, 2 nvriy f rnne, rnne, The Groningen, Groningen, of University 2 , J.Paridaen in zebrafish, which plays an important an plays which zebrafish, in or downregulation of downregulation or 1 , D.Zivkovic 168

compartment size through size compartment 1 , M.Peppelenbosh her5 in embryos in 239 2

Posters Posters 240 in and differentiation, whichweconsideredforfurtherfunctionalanalysis. zebrafish in TFs CA same in invoked not previously the TFs to attention calls analysis expresses our addition, In vertebrates. whichhigher group, prethalamic fish the from e.g. conserved mammals, are to groups some DA all for in differentiation expressed DA universally of as mechanisms far Further, so neurons. identified been have TFs no as brain, the of distinct in regions codes TF distinct by accomplished be may specification DA that indicate data Our mammalian CA groupsbasedonTFexpressioncodes. and zebrafish between ofDAneuronshomology potential establish (3) 2003); Rubenstein, and (Puelles model location the (2)determine CAneurons; within the of longitudinal and transverse TFs subdivisions of the group of forebrain postulated code by distinct the prosomeric combinatorial each a define identifies (1) that to us enabled data analyzed Our was stages. embryos developmental zebrafish four mount at whole of CACNS The resolution. with cell single correlating at analyses territories in forebrain coupled the with anti-TH fluorescent immunohistochemistry in and optimized confocal microscopy an to patterns of conduct co-expression advantage took distinct We differentiation. in expressed genes 60 Based on gene expression data from public databases (www.zfin.or Wullimann, 2002). cell position, their on based shape, distinguished degree of be Tyrosinecan hydroxylase groups (TH) DAimmunoreactivity, Diencephalic and projection hindbrain. behavior (Rink the and to confined are neurons NA while forebrain, the in exclusively groups DA form Zebrafish group. neuron CA each for characteristic code factor transcription the investigate to wellcharacterized. zebrafish Weuse quite been has differentiation Diencephalic DA groupsarelesswellunderstood. and specification during activation TFs of For CADAsome mesencephalic mammalian the like timing groups, sequential groups the A8-A10, and TFs. of specified expression combinatorial be of may prepattern (DA) local brain by control the dopaminergic distinct of under differentiate Hence, regions distinct differentiation. in located neuronal neurons (NA) specifies noradrenergic or events signaling local of Current and behaviors. projection precursors neural in prepattern and (TF) factor transcription of morphologies combination atdistinct a that distinct is form understanding cellular distinct (CA) neurons with of catecholaminergic locations identifies anatomical groups specific brain, developing factors the In transcription specific Dept. ofDevelopmental Biology, InstituteofBiologyI, University ofFreiburg, Germany of A. Filippi, C.Jainok,andW. Driever expression catecholaminergic groupsinzebrafish combinatorial The 169 Neurobiology I:Patterning hybridization protocol hybridization situ in g), we selected approximately expression of expression of differentiation the understanding factor hypophysiotropic The genetic pathways establishing a as acting proliferation andsurvival. Besides neuron controlling effects functions. neurotrophic elicits PACAPalso release, growth-hormone regulating developmental and has sequence physiological Thepeptide teleosts. other various with and neuropeptide pleiotropic PACAPa and is evolution zebrafish during conserved highly been of genomes the in found been have PACAP.the and of (PRP) Twocopies peptide related glucagon the of member peptide superfamily.a The is (PACAP) polypeptide activating cyclase adenylate pituitary The University, Huddinge, Sweden Department of Biosciences and Nutrition, Karolinska Institutet, School of Life Sciences, Södertörn G. Lauter,I.SöllandHauptmann PACAP inZebrafish Neurodevelopment Neurobiology I:Patterning suggests apossiblefunctionforPACAP duringearlyneuronaldifferentiation. of The ganglia. co-distribution cranial in and placode olfactory the tegmentum, in as tectum, well as pretectum, cord spinal tuberculum, rhombencephalon, posterior region, preoptic telencephalon, the of ganglion. trigeminal the in and cord spinal the hindbrain, the found We onward. somitogenesis from detected be can expression mRNAs during zebrafish development. By means of means By development. zebrafish during mRNAs pacap mRNA with that of several key regulators of neuronal development, neuronal of regulators key several of that with mRNA pacap pacap gene encodes for a prepropeptide that is cleaved into a PACAP- pacap cells are only poorly understood. In a first step towards positive cells, we give a detailed description of the of description detailed a give we cells, positive pacap pacap gene, named gene, was present in clusters in present was pacap2 170 to be expressed in expressed be to pacap1 and pacap1 hybridization situ in pacap2, 241

Posters Posters 242 the is with axons motor acetylcholine these receptors of (AChRs) on alignment the precise muscle the surface. One as key well molecular player as in targets this muscle process their to cord spinal the from axons motor of navigation proper involves connectivity neuromuscular of Formation ofCellDepartment and Molecular Biology; University ofPennsylvania, Philadelphia, USA L. Gordon,Jing,M.Granato Neuromuscular synapseformation:anotherrolefortheplanarcellpolaritypathway MuSK ortholog MuSK neuromuscular first the of of sites center the the synapses. prefigure Recently, in and our laboratory manner has accumulate shown that dependent the AChRs secreted MuSK ligand axons, a motor in cells of muscle arrival the before Even cells. muscle of by the nerve-derived ligand agrin, MuSK triggers the accumulation of AChR clusters in the center MS. sn lv cl iaig w ae lo xmnn wehr h subcellular the whether examining during theactivationof‘classical’ frizzled/PCP pathway. also are components these of we relocalization dynamic the mimics components PCP these of of imaging, localization downstream cell fibers muscle live in Using function alsounplugged/MuSK. pathway PCP the ‘classical’ of the components and downstream pathway signaling MuSK the between frizzled/ AChRsare planar similarity and cell of polarity extent pathway axons (PCP) the motor pathway, explore which Towe to have begun fiber to muscle analyze the whether additional of restricted. center the in zone specialized a cascade in muscle cells (Jing et al. 2009). We hypothesize that this signaling cascade establishes 171 muscle specific , and that this interaction triggers a triggers interaction this that and unplugged, inase, MuSK, which is expressed only on muscle cells. When activated When cells. muscle on only expressed is which MuSK, kinase, Neurobiology I:Patterning -dependent signaling dishevelled-dependent wnt11r binds the zebrafish the activityregulatesneurogenesis. expression patterns of the genes regulated by activity in early order to definethe the mechanismin the bydetermine which to want Weactivity now neurogenesis. normal for excitatory necessary are development its of phases and receptor glycine the that suggests This interneurons. classes of several in neurons differentiated of loss major a observed we microscopy, confocal in strychnine (KCC2, cases examining transgenic lines three expressing GFP in the specific cells and In with the use receptors. While neurogenesis. in oferrors majors severalprovoked activity neuronal glycineantibodies of disruption KD), alpha2 and of subunit alpha2 embryonic the of either blocking its action using the specific antagonist strychnine or by the specific ‘knockdown’ all the neurons from the onset of development. We also targeted the glycine receptor directly by in hyperpolarizing glycine rendering gradient, chloride the reversed and over-expressedKCC2 we development, early during transmission glycinergic depolarizing of role the test to order In development. The role of this paradoxical depolarization during early development is unknown. expression of the extrusive chloride and potassium ion co-transporter KCC2 during later stages of delayed the until so remains and high initially is level chloride intracellular the because occurs by mediated depolarization is chloride ions via glycinergic and GABAergic receptor-channels. development This depolarization (excitation) during activity neuronal of forms first the of One depathologie etbiologiecellulaire,Département Université deMontréal, Canada S. CôtéandP. Drapeau Role ofneurotransmission duringneurogenesisinthezebrafish embryo Neurobiology I:Patterning 172 243

Posters Posters 244 do To profiles ofprogenitorcellsindependenttheircellularidentity. in role key a play In a second to approach we use a new likely transgenic line Tg[pcna:EGFP] to are obtain the gene expression which genes neurogenesis andregeneration. investigating zebrafishcurrently the of are lesion We severe a telencephalon. after days 3 cells glia of we profile expression Therefore, gene lesion. the after cells were obtained response proliferative strong glia a shows which brain zebrafish and non-proliferating the in proliferating of profiles obtained. In addition, we are interested in the regulatory gene program underlying regeneration expression gene the this Using cycle. method, cell to the of dye S-/G2-phase DNA in vital were which a cells applied positive GFP we specifically isolate cells, GFP-positive selecting for addition In brains. zebrafish fluorescent activated cell sorting we isolated different cell populations line fromUsing progenitors. adult proliferating transgenic including characteristics, glia radial the and ependymal- with cells that find We [1]. cells neuroepithelial and glia radial both of characteristics show brains vertebrate non-mammalian of cells Progenitor brain. Different types of neural progenitors have been identified in various regions of the adult zebrafish Patterning andRegeneration ofthe Vertebrate Brain, BIOTEC/CRTD, TU-Dresden, Germany D. Freudenreich, S.Hans,J.Kaslin,V. Kroehne,J.Ganz,S.Dudczig,M.Brand Gene expressionprofilingofneural progenitorcellsinadultzebrafish brain D, BrandM. PLoS ONE. 2009Feb andposterStefanHans [3] Temporally-controlled in recombination zebrafish. site-specific Hans S, Kaslin J, Freudenreich Huh TL, Chitnis AB. Dev Biol. 2007Jan embryos.zebrafish systemof SY,nervous Yeo the in signaling HS, M,Notch Kim Kim of pattern [2] Fluorescent protein expression driven by her4 elements regulatory reveals the spatiotemporal 2009 May andposterJanKaslin Neuroscience of M.Journal Brand S,and H,Hans M,Grandel J,J,Niche. Kaslin Geffarth Ganz [1] Stem Cells in the Adult Zebrafish Cerebellum: Initiation and Maintenance of a Novel Stem Cell these progenitors. of potential cellular the revealing label, EGFP the carries progenitors Her4.1 of progeny induced recombination can be observed after the administration of tamoxifen. Subsequently the Tg[her4.1:mCherryT2ACreERT2]. In combination with a red-to-green cells: glia reporter in recombinase Cre of activation controlled temporal the line enables that line transgenic [3] successfully 173 oa fide bona ieg taig f h poey f e41 oiie el, e rae a created we cells, positive Her4.1 of progeny the of tracing lineage Neurobiology I:Patterning Tg[her4.1:GFP] Tg[her4.1:GFP] 2 labels [2] differentiation andmaintenanceofmotorneuronprogenitorsduringembryogenesis. the in the investigating signal for useful be will GFP it and cord spinal and robust hindbrain zebrafish of neurons motor displayed Tg(zccndx:GFP), line, transgenic The development. early suggest results These the cord. spinal that the in unaffected but hindbrain, the in only neurons motor of lost the caused knockdown Gli1 while cord, spinal and hindbrain loss both in the neurons motor in of resulted Dx cyclin zebrafish of knockdown morpholino line, this using By pattern. and placode olfactory cord, embryos. Thereafter, spinal a transgenic Tg(zccndx:GFP) line was and established with identical expression hindbrain of neurons neuronmast supporting cells in zebrafish lateral line by microinjection into one cell of motorzebrafish the in of expression region promoter GFP 4-kb A embryogenesis. during cord spinal and the that indicated hybridization We identified and characterized a novel zebrafish cyclin Dx/zccndx gene. Whole-mount in situ Biology, NationalInstituteofGenetics, Mishima, Shizuoka, Japan; 1 A novel zebrafish cyclin dx gene: expression profile, requirement for the development of for thedevelopment C.H. Cheng requirement profile, expression primordium motorneuron,pmnandcharacterization gene: ofitspromoterregion dx cyclin zebrafish novel A Neurobiology I:Patterning Biological Chemistry, Academia Sinica, Taipei, Taiwan; Institute of Molecular and Cellular Biology, National Taiwan University, Taipei, Taiwan; ee a rqie fr oo nuo poieain n mitnne n the in maintenance and proliferation neuron motor for required was gene zccndx 1,2 , Y-C. Chen 2 , G-D.Chen mRNA was expressed in motor neurons of hindbrain of neurons motor in expressed was mRNA zccndx 2 , C-C.Hung 2 , K.Kawakami 3 Division of Molecular and Developmental 3 , F-L.Huang 174 ee ol drive could gene zccndx 1, *, andC-J.Huang 2 Institute of 245 2, *

Posters Posters 246 expression ofspinalcordcells,theirparticularmorphologyandfunctioninbothspecies. in specific cell types. These will be invaluable tools that will enable us to relate the specific gene as such isolating of using means putative a are cis-elements, which as are we elements, then used populations non-coding to generate conserved interneuron reporter Rohon constructs. identify specific Our to analyses label of approach and genes to bioinformatic a order interneurons In labelled species. that both in transgenics cells Beard generated successfully and system TOL2 of upstream sequence from construct a made we principle, of proof a As during development. spinal cord interneurons. These will be used as genetic tools to study the neural circuit formation of populations different on focusing models, animal both in types cell labelled specifically with well as for physiological studies. With this in mind, we aim to develop a series of transgenic lines and of anatomy simple The stimuli. physical and chemical different in network participate neural and given a specificity, form acquire neurons how understood from far is it yet As *School ofBiologicalSciences, University ofBristol, UK Physiology Development Department, &Neuroscience CambridgeUniversity, UK JL. Juárez-Morales, A. Roberts*andK.Lewis Genetics Tools toStudy Neural Circuit Formation inZebrafish andXenopus 175 Xenopus make them ideal models for the study of neural formation and gene expression, as evx1,evx2, isl1, chx10 , have resulted in reporter constructs that are expressed are that constructs reporter in resulted have lbx1, and Neurobiology I:Patterning with a GateWaya with HuC rerio Danio This study will not only allow us to understand the cellular basis of our phenotype but also but Neurogenesis anddirectionalsignalingbetweencells. phenotype our of basis cellular the understand understand howcellsexploitthetrafficmachinerytoregulatekeyeventssuchas: to us allow only not will study This a moregeneralway. in endocytosis block to and pathway the of steps precise at trafficking with interfere to used be can GTPases small these of forms dominant-active or dominant-negative and Moreover,drugs involved. is SARA which in signaling the on influence their their and neurogenesis in study function their to proteins fusion GFP Rab of advantage behavior live and during cell taking division as well as a imaging morpholino knock-down approach to liveanalyze use will I pathway. my attention. Several small GTPases, the Rabs, mark the different compartments in the trafficking My interest lies on exploring the role of trafficking in neurogenesis with SARA being the center of and thatknock-downbymorpholinosleadstoaneurogenicphenotype. zebrafish SARA show that it can segregate asymmetrically during cell division in the spinal cord in Studies division. cell asymmetric in asymmetric or division cell symmetric in symmetrically segregate either they that is drosophila, in mainly lab our in studied endosomes, SARApositive the these to of feature binds interesting An endosome. it early the which on (PI3P) of phosphatidylinositol-3-phosphate means the by domain FYVE a has it domains functional other to next play: into SARAcomes where is This mediators. signaling intracellular to signal the propagates internalized into early endosomes and signaling is induced after a phosphorylation event, which Smad In our lab one main focus lies on a TGFβ- signaling adaptor protein signaling. called TGFβ- SARA, also and which on Notch- stands NGF-, for for also signaling in endocytosis but for role a suggested membrane evidence more plasma and more years, the the pathway, during signaling at EGF the for happen finding this After endosomes. exclusively early not possibility does the raising transduction view a signal emerged, has that signaling of view new a years 20 almost now For University ofGeneva, Switzerland S. Abke andM.Gonzalez-Gaitan Endocytosis andneurogenesisinthezebrafish spinalcord Neurobiology I:Patterning anchor for receptor activation. Upon ligand binding, the heteromeric receptor complex is 176 247

Posters Posters 248 nervous zebrafish embryonic L. Braeutigam the in neurons 3 Urocortin system and 1 Urotensin of Localization 1 dissect thefunctionoftheseCRHfamilypeptidesinstressinducedlocomotion. now can we mRNA, UCN3 and UTS1 of expression embryonic the on data detailed our With the stressresponseandlocomotionviatheseneuropeptides. are expressed in key neuronal sites regulating locomotor behavior, suggesting the interrelation of immunohistochemistry during zebrafish embryogenesis. We indeed found that UTS1 and UCN3 peptides, Urotensin 1 (UTS1) and Urocortin 3 (UCN3) by whole-mount in situ hybridization and thereby and stress circuitssystem and locomotion, we analyzed the spatial transcript brain distribution of two CRH family aminergic these with interact changing locomotoractivity. To to achieveadeeperunderstandingoftheconnectionbetween known are peptides family CRH are modulated by neuronal the neurotransmitters specific dopamine, noradrenaline on and circuits dependent serotonin These cell. Mauthner the and in neurons reticulospinal including is brainstem all the in circuits vertebrates. Locomotion locomotion. in alterations including patterns behavioral and changes physiological of variety a of body’scomprised The is stress to response Hoegskola, Stockholm, Sweden Biosci. and Nut., Karolinska Institutet, Stockholm, Sweden; 177 1 , J.Hillmer 2 , I.Soll 2 , G.Hauptmann 2 2 School of Life Sciences, Soedertoerns Neurobiology I:Patterning on ES apiue hs ee be son eoe n eprmns r otie to outlined are experiments and before This workwasby supported theCIHRandNSERC. shown been never investigate thisfurther. has amplitudes mEPSC down neural scaling on relevant TNFα of effect behaviorally chronic the Additionally, in development. embryonic alterations during networks compensatory and to fibre contribute may muscle scaling of that synaptic evidence observations new provide activity Our synaptic of synapses. blockade chronic following existing activity swimming strengthen to blockade activity to response in multiplicatively up scale mEPSCs Glutamatergic development. during homeostasis maintaining of means a as vivo in scaling synaptic for role a support results These Conclusions: towards tendency a longer burstdurationsinswimmingepisodesthancontrol. showing activity, muscle increased towards trends have embryos treated CNQX- that show recordings Muscle scaling. synaptic with correlated be may that conditions swim. frequency, could fish CNQX-injected mEPSC Further and analyses of stereotypical swimming AP-5 behaviour including suggest whereas differences between experimental embryos, parameters TTX immotile Behaviourally, in described. Other resulted also treatment are 24hrs. ratio AMPA:NMDA as and interval, glutamatergic little inter-event (decreased) kinetics, as down after scaled significantly amplitudes in mEPSC resulted TNFα of Injection changes. significant, attributed to changes in intrinsic excitability of neurons. AP-5 treated fish showed no significant a ~1.4. by in up resulted amplitudes mEPSC (2-4dpf) Current threshold data shows that increased scalingmEPSC amplitudes in CNQX-treated fish cannot be 48hrs size, for mEPSC glutamatergic CNQX in increase and multiplicative TTX with activity Blocking Results: 28hpf), touchresponse(30hpf),andswimming(2-4dpf)werealsoperformed. neurons and muscle fibres were made at 4dpf. Behavioural analyses of spontaneous coiling (17- acute spinal from mEPSCs of recordings clamp patch Whole-cell control. a as vehicle or application), during amplitude mEPSC increase to known (molecule TNFα blocker), (AMPAR CNQX (hpf) and 48hpf with either TTX (voltage-gated Na (voltage-gated TTX either with 48hpf post-fertilization and (hpf) hours 17 between injected were embryos Wild-type methods: and Materials on earlylocomotorbehaviours. developing the in scaling synaptic investigate little understanding of the relationship between synaptic scaling and behaviour, so our goal is to increase or decrease in the size of miniature excitatory post-synaptic currents (mEPSCs). There is in response to changes in activity, one of the best documented being synaptic scaling, a uniform strength synaptic modify to place in mechanisms homeostatic are There network. neural a form that circuits into wired cells, nerve of populations through generated are Behaviors Objectives: Pathology andCellular Biology, Universite deMontreal, Canada L.D. KnoglerandP. Drapeau Homeostatic synapticplasticityinthedeveloping spinalcordanditsbehavioural correlates Neurobiology I:Patterning (zebrafish) spinal cord and its effects its and cord spinal (zebrafish) rerio Danio + channel blocker), AP-5 (NMDAR blocker), (NMDAR AP-5 blocker), channel 178 249

Posters Posters 250 localization asinhighvertebrates. similar a following adults, in bulb olfactory and ventricle telencephalic the around concentred but development, during brain the throughout distributed are zebrafish in areas neurogenic the that demonstrate results present Takentogether ventricle. telencephalic of tanycytes and cerebellum; cells and ependymal ventricle the to and Rhombencephalic fibres bulb olfactory the the to restricted was of Sox-2 animals adult finally, in cells epithelial in also and localizations the same the in found was dpf 50 and of 25 by opticum; tectum and diencephalon the bulb, olfactory cells scattered in detected also was Sox-2 The dpf 15 dpf). by period; (120 larval the stage during ventricles adult to remain and dpf 40 to brain the lie that cells epithelial the to restricted is it that revealed Sox-2 of immunolocalization up increase then dpf, progressively 30 postfertilization), to (days up 10dpf decrease at maximal are expression mRNA Sox-2 of levels qRT-PCRby larval assessed were stage adult to from immunohistochemistry.and zebrafish that demonstrated Results of brains The undergo. they changes developmental the establish to and zebrafish, of brain the in areas neurogenic potential the identify to aimed was study This brain. which neurogenesis is not restricted to a limited number of areas but is widely distributed in the neural presumptive as used be stem cell marker.could Therefore, we decided to investigate its Sox-2 expression in the brain of zebrafish, in that demonstrated been has recently and organs, These molecules are also involved in the development of both central They are highly nervous phylogenically preserved system and have been and detected from mammals sensory to nematodes. high motility group (HMG box), and which play critical roles in the early stages of development. the of genes by encoded proteins The Oviedo, Spain Celular,de Biología Universidad y Morfología de *Departamento Italia;Messina, di Studi degli e CISS (centro di ittiologia sperimentale per la Sicilia), Facoltà di Medicina Veterinaria, Università Dipartimento di Morfologia, Biochimica, Fisiologia e Produzione Animale, Sezione di Morfologia, A. Germanà,G.Montalbano,MC.Guerrera,E.Ciriaco,J.A.Vega* SOX-2 Expressioninthebrain ofdeveloping zebrafish 179 family are related transcription factors, containing a containing factors, transcription related are family Sox Neurobiology I:Patterning Res., 197:157-165. asymmetries”.left–right anatomical Behav.in Brain. differences show lateralization behavioural Facchin,opposite (3) L.,for F.Argenton, selected rerio Bisazza,Danio A.and (2008). of “Lines concordant brain, andgutleft-rightasymmetries”. heart Development, 127, 3567-3579. Bisgrove, (2) B. W.,J.Essner J. and YostH. J. (2000). pathways“Multiple regulate midline the in formation”. GenesDev., 16, 2339-2344. (1) Burdine, R. D. and Schier, A. F. (2000). “Conserved and divergent mechanism in left-right axis wildpopulationsasanevolutionarystablestrategyforpredator avoidance. rerio the in selected positively been has allele this that Wehypothesize vesicle. Kupffer’s of cilia a possible involvement of this mutation in the generation and maintenance of nodal flow by the right- or bilateral sided) in approximately 50% of (either observed embryos. Current experiments are pattern aim at investigating expression gene their in altered are pathway signaling Nodal asymmetries. left-right diencephalic in involved genes three females’least mutant at of that analysis showed hybridizations progeny and visceral of establishment the for responsible age of the mother. Wethe by investigated affected is in progressively allele this this strain of the expressivity that that expression suggesting of crosses, asymmetries members subsequent of the signaling in diencephalic increases pathways reversed with progeny of percentage a generate variable expressivity. The analyses revealed that maternal in selective recessive most cases a performed these of homozygous hypothesis we mutant effect the mothers mutation. The trait, test reversed to anatomy this is carriers expressed mutant in of the as offspring identified analysis with females a frequency mendelian on that crosses suggests preliminary a on Based structures. allele responsible for the disruption of normal left-right patterning in zebrafish neuroanatomical mutant spontaneous a of isolation the to lead have could lateralization behavioral for selection of frequency the in increase reversed significant left-right a asymmetry had in the mirror epithalamus. a In scrutiny the in present preference study it eye is right proposed the that for Facchin’s In a recent study Facchin et al. (3) showed that the epithalamic regionofthedorsaldiencephalon(2). progeny of lines of zebrafish artificially selected in the zebrafish forebrain and are involved in early establishment of left-right asymmetries in the Moreover, studies have found that Nodal-related signals also asymmetrical, left-sidedactivationofaNodalmediatedgeneexpressioncascade(1). regulate asymmetric gene expression It has been demonstrated that in vertebrates the initiation of left-right patterning depends on the are thoughttoberegulatedbyevolutionaryconservedgenes. and functionally asymmetric. The development of left-right patterning and cerebral lateralization visceral of disposition the organs. This asymmetrical organization extends to the vertebrate in brain that is both anatomically asymmetries left-right distinct displays plan body vertebrate The ofBiology,Department University ofPadova, Italy of developmentA. Domenichini andF. Argenton the in involved allele effect maternal zebrafish left-rightasymmetries a of analysis molecular and Genetic Neurobiology I:Patterning 180 n situ In 251 D. D.

Posters Posters University ofFlorida, St Augustine, USA; 1 252 F. Ono Formation ofspinalnetwork dependentondomain-specific Pax genes National Institutes of Health, Bethesda, USA;Bethesda, Health, of Institutes National transcription factorsinthenetworkformation. newly identified gene trap line enabled classification of spinal interneurons and revealed roles of examined the effect of cord. spinal of the that with of region dorsal the in expressed is Pax8 and theexpressionofpax2,agenecloselyrelatedtopax8wasdownregulated. than smaller were embryos these in vesicles otic Pax8, functional of lack the to Due the embryos. abolished insertion RFP the that showed PCR Real-time midbrain-hindbrain boundary, theoticvesicle,hindbrain,spinalcord,andretina. the of intron first the in occurred insertion RFP the trapped (RFP) protein fluorescent red the which in zebrafish of line involved in diverse aspects of development. Using a gene trap technique, we established a stable temporally.and spatially both expression, gene on control strict a exert factors transcription development, vertebrate During aoaoy f oeua Pyilg, ainl nttt o Achl bs ad Alcoholism, and Abuse Alcohol on Institute National Physiology, Molecular of Laboratory 181 1 , T. Ikenaga Pax8. is expressed in CiAs, while CiAs, in expressed is Pax2 1 , J.Urban pax8 and/or 1 , N.Gebhart Pax genes comprise an important family of transcription factors transcription of family important an comprise genes pax2 knockdown in 3 National InstituteofGenetics, Mishima, Japan 2 , K.Kawakami 2 The Whitney Laboratory for Marine Bioscience, Marine for Laboratory Whitney The gene. RFP expression is detected in the in detected is expression RFP gene. pax8 is expressed in CoBLs and IpLos. We IpLos. and CoBLs in expressed is pax8 pax8-expressing interneurons. The use of 3

Neurobiology I:Patterning xrsin atal overlaps partially expression Pax2 expression in homozygous in expression pax8 pax8 gene. In this line, this In gene. AMIGOs (amphoterin-induced Center,Neuroscience University ofHelsinki, Finland X. Zhao,J.Kuja-Panula,P. Panula,H.Rauvala The roleofamigo1inthedevelopment ofzebrafish earlybrain’s catecholaminergic system Neurobiology I:Patterning h 5p c-netd ave vn hw yeatv efcs n h bhvoa ts. u study functions invertebratecatecholaminergic systemdevelopment. Our test. behavioral the in effects shows an important hyperactive role of AMIGO1 in the developing show zebrafish larval brain and corresponding even larvae co-injected 5dpf The oligos. morpholino mRNAwith of AMIGO1 together co-injection by successfully rescued been catecholaminergic system deterioration. The the AMIGO1 knock-down morphant phenotypes have of consequence expected an is which morphants, 5dpf AMIGO1 in distance decreased to Furthermore, the swimming behavior test shows a disorganized pattern of swimming in addition development. of beginning very the from detected be could which impaired, also are eyes and hindbrain. TH cell cluster formation and distribution in the morphants’ telencephalon, midbrain the in projections fiber neuronal TH-immunoreactive in organization disordered and decrease (TH) immunohistochemistry for labeling zebrafish catecholaminergic system shows a prominent in 3dpf(days Tyrosineshow expressed. hydroxylase normally usually is AMIGO1 where brains areas the at larval defects development knock-down mount immunostaining AMIGO1 larvae. injected andwhole 5dpf and fertilization) by western-blot past both confirmed been has in the zygote by the introduction of specific morpholino oligos via microinjection. The inhibition AMIGO1 roles during early development stage, we have managed to knock down its expression explore to order In found. far so been have functions [Tomiofew vitro 2003], in O, neurites of 2009]. H, But except for its potential [Shunsaku roles in the apoptosis of stage granula neurons and mediating fasciculation development early during system nervous central end the in expressed glycation mainly be advanced to found and hybridization situ for in mount whole by detected been (receptor has embryo mice RAGE by induced K.P.,[Juha be amphoterin the with in ligation of AMIGO1 products) pattern expression The 2003]. could AMIGO1 of Expression of transmembrane proteins that belong both to the LRR (leucine-rich repeat) and Ig superfamilies. gene and orf) form a novel family (AMIGO1, AMIGO2, AMIGO3) 182 253

Posters Posters 254 Golgi localizationinPurkinjecells. is disrupted. We will further discuss the role of aPKC in the polarized dendrite formation and the cells that (aPKC) Purkinje PKC atypical of function the which in that mutant the suggesting in dendrites primary multiple retained found also We polarization, formation. dendrite morphological polarized the in preceded regulates dendrite Golgi localization localized primary of Golgi root and the at cells localized Purkinje exclusively was apparatus Golgi that found We morphology. and neurites only one multiple of them had was finallyinitially selected asthey the futurethat primary dendriterevealed to formcells the maturePurkinje polarized of changes morphological Timethe cerebellum. for mammalian observations in lapse observed as dendrite primary single antero- along location somal posterior and the midio-lateral axes. In spite of the morphological diversity,to all Purkinje cells according have a morphologies dendritic different showed cells Purkinje cells and in observed their specifically morphologies at Venus membrane-targeted single cell express resolutions. We that found fish that transgenic zebrafish Purkinje established We the process. control that mechanisms the and morphology polarized highly the acquire cells Purkinje how single a having studying morphology are we experiments, Taking live-imaging dendrite. for primary zebrafish polarized of advantages characteristic exhibit they since issue this studying for system model good a initial us give the cerebellum the as in cells body) Purkinje patterning. cell dendritic for a process from directly extending controls (dendrites that dendrites mechanism primary the of on number focus the we dendritic branching, that of or processes growth Among mechanisms dendritic understood. as morphology, be such patterning, to dendriticstill are dendrites of of shapes diversity specific pattern remarkable display neurons Although Kobe, Biology, Developmental for Center RIKEN Formation,Japan Axis zebrafish Vertebrate for the Laboratory in cells Purkinje K. Tanabe, of YK. Bae,S.Kani,T. Shimizu,M.Hibi formation dendrite cerebellum polarized controlling Mechanisms 183 Neurobiology I:Patterning f h atvt i te eta poeios ht nue hi sic fo pouig ern to neuron forOPC producing from required switch their activity induce that ofShh oligodendrocyte inthevertebratespinalcord. progenitors ventral the in activity regulator Shh of is apositive modulation temporal a triggers Sulf1 that propose We cord. spinal ventral the Sulf1 in specification that suggest findings These that productionofventralneuronsisnotimpairedwhereasOPCareabsentinsulf1morphants. of Shh target genes is downregulated in the ventral neural tube. Our preliminary results indicate tested We 2006). al., function in zebrafishet by morpholino loss(Danesin of function experiments.signalling ligands In sulf1 and morphants, expression of distribution extracellular activity Shh and regulates transport that Sulf1 that indicate chick in studies shown function of Gain HSPG. on depends have family Hedgehog from Vertebrates and their Drosophila andtherefore from Studies ligands extracellular for affinity activity inregulatingsignallingpathways. their modulates pattern of modification sulfation This surface. HSPG cell the at (HSPG) Proteoglycans Sulfate Heparan from position specific at groups sulphate hydrolysing enzyme an for encodes Sulf1 results). present 2006, al., et (Danesin theventral zebrafish and chick including into vertebrates, among conserved strongly is expression expands later and floorplate the in neural progenitors just prior to production OPC specification in this domain. This neuronal spatio-temporal pattern of of waves main the oligodendrocyte of subpopulations. specification The sulfatase Sulf1 is potential player involved in these events. Indeed, its expression starts after neuronal the different induces underlying thistemporalchangeofneuralprogenitorresponsetothesamesignal. generating mechanisms Shh molecular the of question the raising domains cord, spinal production, ventral the in (OPC) precursors neural neuronal distinct to Subsequently of formation the Hedgehog (Shh) morphogenetic gradient, originating from notochord and floor plate, establishes model to study the emergence of the diverse neural populations. In the ventral region, the Sonic system, arise from neural progenitors during embryonic nervous development. central The adult the spinal composing cord oligodendrocytes) is and (astrocytes a cells simple glial and Neurons Centre deBiologieduDéveloppement, Université Paul Sabatier, Toulouse, France C. DanesinandSoula spinal cord ventral the in specification oligodendrocyte Shh-dependant in function Sulfatase1 of Analysis Neurobiology I:Patterning 184 sulf1 255

Posters Posters 256 critical are Nevertheless, contact ventricular common determinantstomaintainneuralstemcellactivityinvertebrates. characteristics. and polarization glial distinct as cells innon- than such properties neuroepithelial epithelial stem retained neural more adult show view, vertebrates the predominant mammalian to contrast In profile. transcriptional ofthe sorting cell activated fluorescent their analyse and progenitors cerebellar isolate specifically used to line transgenic Tg(nestin41:egfp) we cells stem cerebellar regulating mechanism in molecular the involved understand further activity. To signalingleads cell stem ofFgf of reduction significant Inhibition to Fgfreceptor. dominant-negative an inducible with zebrafish used we factors, growth Fibroblast factors, important of family one of requirements the Totest activity.cell stem of regulators important be to thought are ventricle the in propagated Factors stem its and niche the Importantly,cells still remain in growth. ventricular contact through a tissue previously unknown derivative of the and ventricle. movements morphogenetic involving process two-step a in progenitors lip rhombic and ventricle of displacement coordinated the by glia but instead retain neuroepithelial characteristics. The cerebellar stem cell niche is generated typical not are cells Remarkably, stem cerebellum. the zebrafish adult the in niche distinct a in vivo brainthan in lines, of the parts other in we studied the origin and architecture of a previously found unknown stem cell niche using transgenic also and zebrafish Totelencephalon. vertebrates, in in niches cell stem neural of characteristics common understand abundant are niches cell stem these how understood mammals, to not contrast In maintained. is are they how It and development during niches. formed are niches special in place takes neurogenesis CNS, adult the In Biotech. center/Center forRegenerative Therapies, Technical University, Dresden, Germany J. Kaslin,Ganz,D.Freudenreich,M.Geffarth,H.Grandel,S.HansandBrand niche cell stem novel a of maintenance and initiation cerebellum: zebrafish adult the in cells Stem 185 imaging and marker analysis. We show that bi-potent stem cells are maintained are cells stem bi-potent that show We analysis. marker and imaging Neurobiology I:Patterning Institute of Biology I, University of Freiburg, Germany; 1 N. Tiso development of analysis comprehensive A Neurobiology I:Patterning Puppa Pathology and Veterinary Hygiene, University of Padova, Agripolis, Legnaro, ; Italy Olig functioninneuralidentityandglobalshapingofthevertebrate CNSdevelopment. of model regulatory a assemble to helping are conditions pathological neural specific on PCR districts glial of or the nervous neuronal system. in In parallel, proteins microarray-based fluorescent transcriptome expressing analysis and lines real fish time transgenic with weare crossed mutants, decisions, fate cell neural specific in members performing currently Olig of role the detail in evaluate To neural differentiation. findings for Our role a suggest CNS. embryonic the of axis dorso-ventral the along signals Hh and BMP by and and FGF signaling during gastrulation and early somitogenesis, by RA signaling in the hindbrain, domains of the spinal cord. spinal the of domains of expression differential observed we other members display stratified expression in diencephalon, hindbrain and spinal cord. Notably, According to our data, Three specification. type cell the of members The Unit, ofPadova, Hospital Italy and olig3 zebrafish Interestingly, CNS. the in regionalization and induction their control pathways analysis of global a Weperformed (olig4). have anamniotes in one fourth a and models, vertebrate known eatet f ilg, nvriy f aoa Italy; Padova, of University Biology, of Department olig4 together establish most of the expression domains corresponding to mammalian 4 1 , W. Driever , A. Filippi olig genes during zebrafish embryonic development and determined which signaling genes in CNS patterning, as well as in multiple cell fate decisions during decisions fate cell multiple in as well as patterning, CNS in genes olig 2 2 , F. Benato , F. Argenton gene family encode for bHLH transcription factors involved in neural in involved factors transcription bHLH for encode family gene Olig knock-down/over-expression experiments and characterizing genetic characterizing and experiments knock-down/over-expression olig olig1 is specifically confined to the oligodendrocyte lineage, whereas the olig2, genes expression, regulation and function in zebrafish CNS zebrafish in function and regulation expression, genes olig 1 genes (Olig1, genes Olig , E. Negrisoli 1 and olig3 genes within specific motoneuron and interneuron and motoneuron specific within genes olig 3 , N. Modena expression appears to be regulated by Nodal by regulated be to appears expression olig4 and Olig2 3 Department Department of Public Health, Comparative

2 eatet f eeomna Biology, Developmental of Department 1 , E. Vaccari ) have been identified in all in identified been have Olig3) 186 1 , S. Campanaro 4 Neurosurgery Neurosurgery 1 , A. Della Olig3. 257

Posters Posters 258 live imagingtechniquestoinvestigateOTcircuit formationandfunction. with along modulations genetic employs study ongoing The system. hypothalamo-hypophyseal the of establishment the follow and derived are precursors OXT which from area diencephalic fluorescently labeled OXT neurons. Using two-photon microscopy we could trace the primordial specification existin of OXT neurons circuits in zebrafish we OXT have generated which a transgenic OXTunclear reporter line with itremains zebrafish and if so what dictates their formation. To However, study the mechanisms of differentiation and mammals. and zebrafish conserved between are decisions fate cell OT in involved factors critical that indicate studies us Secondly,and mammals. others by in neurons of thousand of tens to containing compared cells as of neurons, clusters OT larger 15-20 merely of cluster cell single a contains hemisphere brain neurons OXT and of their circuitspecification formation. the underlying Firstly,processes cellular zebrafish haveand amolecular relativelyexact simple OXTthe system paradigm understand – excellent to each an as sexual serve Zebrafish bonding, pair responses. stress trust, and social anxiety in arousal, and involved behavior is OXT system, nervous central the In periphery feeding. the in targets control cells OXT through the hypothalamo-hypophyseal system, neurotransmitter: where it was found to facilitate labor and and breast- hormone both as fish] functions in OXT (itnp) mammals, In isotocin hypothalamus. vertebrate the and of cells of (oxtl) population important an oxytocin-like constitute as known [also neurons (OXT) Oxytocinergic Molecular Cell Biology, The Weizmann InstituteofScience, Rehovot, Israel J. Blechman, A. GutnickandG.Levkowitz Specification andcircuit formationofoxytocin–secretingneurons 187 Neurobiology I:Patterning characteristics ofCiAs. unique the specifying in factors transcription these CiAs. of functions in the analysing are upregulated we Currently specifically are that factors transcription identified have we comparison profiles of these cellsRNA to the trunk cellscompared athave We the same neurons. stagedifferentiated usingmost Affymetrix’sin microarrays.expressed Throughis this GFP where embryos GFP-positive cells from Tg(pax2a:GFP) embryos where GFP is expressed in interneurons. CiAs Work and from Tg( huC:GFP) our lab shows glycinergic- that ipsilateral CiAs, the like of V1 cells, all inhibition in zebrafish spinalexpress cordprovide and are probably functionallyPax2. homologous to mammalianthat V1 We interneurons have of FAC class sorted multifunctional a are CiAs University ofCambridge, UnitedKingdom G. A. Cerda,K.Lewis circumferential specifying in involved factors ascending spinalinterneurons transcription of function the Characterising Neurobiology I:Patterning 188 259

Posters Posters 260 to directlyanalysethepotentialofBMP signallingtoinduceearlytelencephalicmarkers. the in local gain and loss decisions of function experiments performing in the diencephalic anlage and fate currently perspective telencephalon are distinct we inhibiting Hence, regionalisation. or forebrain promoting of either process is plate neural rostral the across activity signalling BMP graded radially a Wethat markers. predict (flh) diencephalic and foxg1) (emx3, telencephalic of expression proper for stages gastrula early and dome between crucial is activity signalling BMP that revealed development of stages several at receptors, I type BMP of the telencephalon in zebrafish. Experiments using dorsomorphin, a competent inhibitor of the the temporal-spatial requirement of BMP signalling activity for the induction and early patterning activity, while diencephalic factors are expanded and radialized. In this project we are in assessing reduced strongly or absent is telencephalon the However, in expanded is plate neural the zebrafish induction in the and fates for neural requirement anterior of basic a is activity signalling BMP of inhibition the that showed network molecular Proteins (BMPs) which the are known that to in be a key evidencerole regulators of neural vital fates. is Previous data a in there play inducing telencephalon. In this context we are analysing antagonists forebrain, the impact of the Bone Morphogenetic Wnt rostral to the additionally factors of signalling establishment the most for the an be requirement of to shown one important been has to inhibition Wnt rise Although CNS. gives vertebrate the and in forebrain structures complex anterior the form evaginates telencephalon The London, College King’s House, Hunt’s New Neurobiology,United Kingdom Developmental for Centre MRC H. BielenandC.Houart pre- and early at gastrula stagesinzebrafish plate neural marginal the in fates cell induce to required is signalling Bmp 189 Neurobiology I:Patterning embryos deprived of BMP of deprived embryos these mutants. bmp2b/swirl bmp2b/bmprI Xenopus disruption of Dynein function in both axons and Schwann cells can contribute to peripheral to contribute can cells Schwann nerve disorders. and axons both in function Dynein of disruption that possibility the raise and maintenance and development nerve peripheral in Dynein for role new potential a reveal data our Therefore, myelination. and wrapping axon for cells Schwann that indicate data preliminary Our analysis. mosaic genetic a if Todetermine mice. and humans zebrafish in disease nerve peripheral to linked been previously has a late step in myelination. Loss of Dync1h1 function in neurons and disruption of axon transport that suggesting larvae, and embryos mutant of roots motor at mutants have very few cells that express myelin genes. Nevertheless, Schwann cells are p resent gene (dync1h1 gene the in mutation insertional an for homozygous larvae that found Surprisingly, we deficits. myelin in result that mutations analyzes lab our axons, around neuropathic pain. To investigate the mechanisms that control the and formation of temperature the myelin in sheath change or pain sense to inability an degeneration, neuron motor paralysis, target axons. The failure to form or maintain peripheral myelin can result in muscle weakness or ensheath and to migrate that crest neural from produced cells glial cells, Schwann by formed is of conduction salutatory myelin PNS, the In rapid PNS). (CNS, systems nervous peripheral for and central the in potentials required action is axons around sheaths myelin of formation The Pediatrics, University ofColorado Denver, Aurora, USA M. Langworthy, B. Appel The roleofDyneincytoplasmic 1heavy chain 1inperipheral nerve myelination Neurobiology I:Patterning function is required in axons or Schwann cells for myelination, we initiated we myelination, for cells Schwann or axons in required is function dync1h1 hi3684Tg ) have myelin deficits in both the CNSA and PNS. In particular, In PNS. and CNSA the both in deficits myelin have ) dynein cytoplasmic 1 heavy chain 1 chain heavy 1 cytoplasmic dynein function is necessary for necessary is function dync1h1 function is necessary in necessary is function dync1h1 190 dync1h1 261

Posters Posters 262 interactions betweenNrp1aandSema3aa,Sema3ab,orSema3d. 1 may have antagonistic roles in central axon guidance. We are currently investigating potential with 22% compared in embryos expressing errors, Nrp1a-GFP alone. with axon These results suggest central that injected Sema3h and showed Nrp- neurons embryos of Nrp1a-GFP, In 63% and alone. morpholino Sema3h expression Nrp1a-GFP exogenous with seen phenotype the the absence of specific Semaphorins. Surprisingly, we found exogenously that Sema3h knockdown enhances embryos in Semaphorin in occur can effect overexpression Nrp1a the each whether ask to spinal cells RB in Nrp1a-GFP expressing knockdown the to to lateral morpholinos just used somites have the We in cord. expressed are that Sema3ab) and Sema3aa (Sema3h, examining potential interactions between Nrp1a and Sema3d or three other Class 3 Semaphorins are we Semaphorin, Toparticular a cord. by spinal caused the is within effect this whether test pathways aberrant take and ventrally grow axons central the cells, other In skin. the into grow axon pathfinding. The most remarkable effect is that some central axons exit the spinal cord and its localization. Interestingly, we found that exogenous Nrp1a-GFP leads to errors in central examine RB to initially cells, RB in construct fusion 1a-GFP Neuropilin a of expression drove We axons. RB of guidance the in receptors Semaphorin potential of roles the investigating are We spinal cord. central axons. These results suggest that Sema3d may act RB to repel the peripheral peripheral axons out in of the reduction affect not did a but axons peripheral repelled Sema3d caused expressed ectopically Furthermore, axons. plate, roof a cord Sema3d, spinal of the knockdown in that expressed showed signal lab guidance our in work Previous periphery. the to exit axons difference between the RB axons is that the central notable axons most remain the in Perhaps the behaviors. CNS different while these the controlling peripheral mechanisms the understanding Wein contact. interested on are another one repel skin the within extends axons Peripheral skin. the and to cord spinal the exits axon, central the off directly extend branch a as axons arises which axon, central The behaviors. different distinctly longitudinally axons, within the display spinal of cord and fasciculate types with that one another.two In peripheral, extend contrast, the peripheral and and cord central spinal the in reside neurons sensory (RB) Rohon-Beard Madison, USA of ZoologyDepartments and Anatomy, and Genetics Training Program, University of Wisconsin- N. S. Asuri andM.C.Halloran central rohon-beard by exit axons cord spinal aberrant elicit interactions Semaphorin-neuropilin 191 Neurobiology I:Patterning A.E. Reyes nttt fr udmna ad ple Booy Snig, Chile; Santiago, Biology, Applied and Fundamental for Institute L. Function ofthehistoneMacroh2avariant intheembryonic development ofthezebrafish Neurobiology I:Patterning 1 we have analyzed the effect the knockdown of Macroh2a2 on the expression of development embryonic in defects in the Later head formation characterized by development. microcephalia and microphthalmia. of Additionally, induces Macroh2a2 of function of fail stages The tail. and region head the in expressed is Macroh2a2 early Macroh2a2 at zebrafish ubiquitously that expressed show is human from anti-macroH2A2 using Immunohistochemistry only variants, RT-PCRthat using show histone two cloned have We participation a In have this work could we macroH2Astudy the that expression of suggests Macroh2a during theembryonicdevelopment. during which the embryonic differentiation, development in of cell participates zebrafish. and of genes specific processes of function the regulate can macroH2A which in evidence essential exist Additionally, not adults. The in be mainly expressed would is MacroH2A2. that because development, protein embryonic and the a during is MacroH2A1 macroH2A1 named that indicates isoforms, MacroH2A1 of two knockout have variant MacroH2A Histone epigenetic different by variants intothechromatin. reversed be can which mechanisms, repression, such as post-translational modification of massive the histones and incorporation transcriptional of histone a causes nucleosomes DNAinto the of assembly The DNAsequence. the in changes without organism, the of multicellular a originate to differentially genome their express regulation zygote totipotential a allows that strict a requires that process signaling complex pathways that participate highly in the cell a differentiation and a is basal stage of development gene repression, Embryonic delaCellule,Moléculaire deLyon, EcoleNormaleSupérieure France. CONICYT C06B03. ECOS- and A.M to 1090416 M.A., to 1070358 A.E.R., to 1095128 FONDECYT by Supported genes. OurresultssuggestthatMacroh2a2couldbeinvolved inheadformation. Departamento de Ciencias Biológicas, Universidad Andrés Bello, Santiago, Chile;Santiago,Bello, Andrés UniversidadBiológicas, Ciencias de Departamento Guajardo 1,2 1 V Muñoz V. , 1 E Barriga E. , mRNA is expressed during embryonic development. embryonic during expressed is mRNA macroh2a2 1 P Bouvet P. , and macroh2a1 3 M Alvarez M. , from zebrafish. Studies zebrafish. from macroh2a2 1,2 A Molina A. , 192 3 aoaoi d Biologie de Laboratorie 1,2 dlx3 and MI Vera M.I. , 2 Millennium pax2.1 263 1,2 ,

Posters Posters 264 and gastrulation during established is prethalamus The (2007) C.regionalization.influences diencephalic PLoS Biol. 5(4):e69. Houart, and N. Staudt, gene functionforspecificcandidates. assess and activities pathway restricted of areas identify fate-mapping, by defined populations transcripts and verifying the data. We have enriched started diencephalic to of allocate list gene the expression analysing patterns are to Currently,we the midbrain. cell presumptive the of this RNA-profiled cells of the diencephalic anlage at bud stage and compared their transcriptome to 2007). In an effort to unravel the genetic components of this specification, we have isolated and are already functionally specified and irreplaceable as early as the bud stage (Staudt and Houart, distinct cell populations. Surprisingly, it was also found that cells of the prospective prethalamus these diencephalic subterritories. This study demonstrated that very little mixing occurs between to raise The give that cells of origins plate neural the reveal to helped has lab our in diencephalon. fate- study mapping recent A pretectum. the caudal-most the and epithalamus, and positioned dorsally the eyes, thalamus, the prethalamus, the telencephalon, hypothalamus, located ventrally the into the subdivided further be early can into latter During hindbrain. behaviours and morphogenetic mid-, fore-, and specifications the type cell of into sequence complex caudal a through develops to forebrain the rostral neural somitogenesis, the from as divided known cells is neuroepithelial It of sheet plate. simple a from arises brain vertebrate The MRC Centre forDevelopmental Neurobiology, King’s College London, UnitedKingdom K.M. Kuerner andC.Houart Dissecting geneticcomponentsofdiencephaliccell-fatespecification 193 Neurobiology I:Patterning knockdown of the of knockdown Morpholino hpf. 48 approximately at starting forebrain zebrafish the of cells most in observed brain development by Morpholino knockdown. Co-expression of reporter transgenes and enhancers which are active in the forebrain and tested the role of the ofthe three with lines role zebrafish transgenic generated have we Toissue, unknown. this investigate the zebrafish In mice. of Dlx genes play an important role in the differentiation and migration of GABAergic interneurons acid glutamic decarboxylase (Gad), the enzyme necessary for GABA production. Therefore, the characterized due to exencephaly. Ectopic expression of via mechanisms cross-regulatory to their due regulatory elements. The forebrain, forebrain phenotype of the in Also, absent cortex. entirely thealmost is in expression interneurons GABAergic of reduction to leading mice mutant nefrs ih proper with interferes migration of immature interneurons is blocked in the ventral telencephalon of telencephalon ventral the in blocked is interneurons immature of migration Single cortex. overlying the to tangentially migrate will and telencephalon ventral the of zones proliferative the in born and hippocampuswhicharenecessaryforinformationprocessing.GABAergic interneuronsare aminobutyric acid (GABA) in the forebrain. These neurons provide inhibitory signals to the cortex The expression of the Biology,University ofOttawa,of Canada Department (CAREG), Genomics Environmental in Research Advanced for Center R. MacDonald,M.Debiais-Thibaud,andEkker Role ofthedlxgenesinGABAergic interneuronandforebrain development ofzebrafish Neurobiology I:Patterning This work is supported byThis workissupported CIHRMOP14460andNSERC. interneuron differentiationandmigrationinvertebrates. the implicate results appears pallium genes throughout forebrain development, comparable to the one described in the mouse. the These to telencephalon interconnected an implies ventral This the from neurons expressing genes causes a reduction in the activity of activity the in reduction a causes genes dlx genes in an evolutionarily conserved pathway controlling GABAergic controlling pathway conserved evolutionarily an in genes dlx Dlx homeobox genes is closely linked with interneurons expressing gamma- expression in the zebrafish forebrain. Also, the migration of the migration Also, forebrain. zebrafish the in expression gad mutant mice show subtle phenotypes if any at all. However, the However, all. at any if phenotypes subtle show mice mutant Dlx cascade potentially crucial for the proper expression of expression proper the for crucial potentially cascade dlx ee i GBegc nenuo dvlpet is development interneuron GABAergic in genes dlx Dlx5/Dlx6 double mutant mice can not be Dlx is sufficient to induce expression of regulatory elements and elements regulatory dlx 194 dlx transcription factors in Dlx/Dlx2 dlx-dependant. Dlx5/Dlx6 double gad is - gad 265 dlx dlx

Posters Posters 266 Rev 8, 819-831. Li, Q., Lee, J.-A. Black, D. L. roundabout (2007). Neuronal zebrafish inaxonregulation homologs suggestroles guidanceandcellmigration.of Dev Dyn221,alternative three 216-230. pre-mRNA splicing.of Nature expression and Cloning (2001). C.-B. Chien,Ray,J.-S., R.,Lee, Robo2 isoformsinzebrafish andrat. Dev GenesEvol 216, 555-563. Dalkic, E., Kuscu, C., Sucularli, C., Aydin, I. T.,58, 325-332. Akcali, K. C., Konu, the O. (2006). of repulsion.to Neuron attraction Alternatively from splicing switch midline governsspliced the receptor axonguidance Robo3 Alternative (2008) Tessier-Lavigne, M. L., Ma, H., Long, B., B. Gore, Z., Chen, is notcrucialforretinalaxonpathfindingbutmayaffectthe pathfindingofCoPA neurons. 23 exon that suggest morpholino this from results Preliminary hybridization. situ in short-probe are able to effectively force exclusion and of (CoPA) neurons ascending primary commissural olfactory sensory neurons, whose axon pathfinding is known to require cells, ganglion retinal in exons spliced are now using splice-blocking morpholinos to test the functional importance of these alternately contrast, the expression profile in non-retinal tissue does not change significantly over time. We In pathfinding. active during development, of stages later at tissue eye in skipped preferentially different at tissue developmental stages: 24, 36, non-retinal 48, and 72 hours versus post fertilization. We retinal find thatzebrafish several exons are in 31 and 29, 23, arepreferentially 19, 7, exons that Robo2 of presence or absence exons the RT-PCR,tested Using we Robo2 29. and several 23, 19, 7, identified exons including spliced, have lab our from results Preliminary 2008). al., et (Chen navigation axon commissural in steps distinct for required are Robo3 of forms two mice in instance, For points. choice different at used be might Robo2 of domains intracellular celldifferentiation, ganglion Different tectum. retinal optic the to pathway their along points Following choice many encounter orlarvae. axons retinal of in embryos tissues non-neuronal not most but in zebrafish, 27 et adult exon (Dalkic Robo2 reported expresses been splicing has alternative it that Indeed, 2006) stages. al., developmental different or at tissues tissue different same in utilized the be in could Robo2 the of to forms navigation splice axon retinal different Conceivably,guides Slits tectum. to binding its retinal and zebrafish 2001), in al., present et Robo (Lee cells only ganglion the is Robo2 zebrafish, In Robo3. and Robo2, Robo1, include isoforms different express to known are that receptors guidance Axon 2007). al., et (Li diversity, and has been implicated in cell-fate determination, axon Alternative splicing guidance, is actively induced and during neuronal development, contributing tissue to proteomic patterning City, USA Lake Salt Anatomy,Utah,& of Neurobiology University of Program, Department Neuroscience MY. Law andC-B.Chien Spatial andtemporal regulationofrobo2splicevariants 195 robo2 exon 23 and confirmed the results by RT-PCR and Neurobiology I:Patterning . For robo2 instance, we 1 2 B. Simões development Investigation of Neurobiology I:Patterning zebrafish, a zebrafish, In differentiation. cell T and neurons sensory of neurogenesis differentiation, experiments epithelial gastric for critical is function Runx3 and development tooth and bone for essential of is Runx2 hematopoiesis, of gain and embryonic early in development decisions and fate ablation cell tissue of differentiation regulators - important Geneas blood, proteins neurons, three all and established bone. Runx3/AML3/Cbfa3. Runx1 is involved and in regulation Runx2/ Runx1/AML1/Cbfa2, AML2/Cbfa1 comprises factors transcription of family (runt-related) Runx The ). by(SFRH/BD/38083/2007) andNCissupported aposdocgrant(SFRH/BPD/9451/2002). support. financial for CCMAR and FCT Acknowledgments: markers. specific cell-type neuron sensory with combination in function Runx3 of knockdown mediated Runx3 of roles possible the of the developing pectoral fins and cranial sensory ganglia. We are now focusing on investigating ganglia, Rohon-Beard neurons and the lateral line primordial. that splicing alternative by generated from dual variants promoters, P1 and P2. Our characterisation of the multiple encodes and exons six contains gene This of diversity the characterised Wehave importance ofdifferentisoformsunknown. (DRG) ganglion relative the rootand unclear is neurons dorsal sensory zebrafish in Runx3 developing of role precise of the but neurons, subpopulations distinct in expressed is runx3 that indicated have chick, and mice in studies Recent reported. studies functional no but described Different splice variants, encoding 2 different protein isoforms, runx3-Shorter and –Longer, were Centro de Ciências do Mar, Universidade do Algarve, Campus de Gambelas, Faro, Portugal;Faro, ofBiology& Biochemistry,Department Gambelas, University ofBath, UnitedKingdom de Campus Algarve, do Mar,Universidade do Ciências de Centro runx3 is dynamically expressed in developing embryonic neural tissues, including trigeminal 1 , N.Conceição runx3 runx3 orthologue was identified and its embryonic expression partially described. partially expression embryonic its and identified was orthologue expression pattern and function in zebrafish peripheral nervous system 1 , M.L.Cancela in peripheral nervous system development, by using morpholino- using by development, system nervous peripheral in transcript variants and encoded protein isoforms. protein encoded and variants transcript runx3 1 andR.N.Kelsh 2 runx3 expression patterns confirms runx3 s upre b a h grant PhD a by supported is BS 196 is also expressed in the base 267

Posters Posters than their zygotic counterparts. Pigmentation and eye growth phenotypes in zygotic in phenotypes growth eye and Pigmentation counterparts. zygotic their than zygotic requirements for requirements zygotic and maternal critical demonstrate results these Takentogether, microphthalmia. in results this In the absence of ATP pathway activity, S-phase is is prolonged, it cell cycle exit that is compromised reveals and pathway required for proliferative retinoblasts to synthesis complete ATPS-phase of the cell cycle the in a normal duration. of fromdeficiencies analysis results Further pathway. synthesis ATP andmicrophthalmia an in pathway, synthesis a GTP in deficiencies from result defects pigmentation pathways: biosynthetic separable to ascribed be can mutants paics pigmentation, respectively. and they identify independent functions for ATP and GTP pathways in mediating eye growth and for maternally derived developing eye. Maternal-zygotic and maternal-effect mutants demonstrate a also critical microphthalmic, requirement resulting from defects in cell cycle exit Zygotic mutants. the in distributions and of amounts normal in present proliferative retinoblasts and within absent, the completely are cells pigment that almost fact the despite decreased, significantly are is pigmentation melanin-derived pigmentation iridophore and xanthophore where defects that catalyzes steps enzyme 6 and 7 bi-functional of this process. a Zygotic synthetase, succinocarboxamide phosphoribosylaminoimidazole 268 and that mutations synthesis, zebrafish recessive monophosphate inosine (IMP) of 5 and 3 two 2, steps catalyzes that enzyme tri-functional of a synthetase, analysis well been an formyltransferase,phosphoribosylglycinamidesynthetase,phosphoribosylaminoimidazole not report have development we affect embryonic cellular Here vertebrate and biochemical during characterized. numerous functions for critical their are processes, signaling purinergic and purines While Molecular Cell andDevelopmental Biology, University of Texas at Austin, Austin, USA J.M. Gross, A. Ng,R.A.Uribe,L. Yieh, R.J.Nuckels in vertebrate pigmentationandoculardevelopment in mutations Zebrafish 197 uie synthesis, purine novo de gart gart GARTand de novo de and paics paics, as these mutants show more severe developmental defects purine synthesis during vertebrate embryonic development, embryonic vertebrate during synthesis purine identify critical roles for roles PAICScritical identify gart gart noe popoioyaiomdzl carboxylase, phosphoribosylaminoimidazole encodes and paics. gart gart paics. gart gart and Neurobiology II:Sensoryorgans paics noe phosphoribosylglycinamide encodes mutants present with pigmentation purine synthesis NOVOpurine DE gart gart and mutants are mutants paics gart gart and network via a rostral loop. This helps define an elementary circuit that is modified bythe ismodified that circuit elementary an define helps This addition ofsensoryinputs,resultinginbehavioraltransformation. loop. rostral motor a contralateral via the to network interneurons and neurons that sensory connect demonstrates synapses glutamatergic analysis after only Electrophysiological arise zebrafish embryonic early in responses cord. touch that Weconclude stimuli. tactile ipsilateral spinal to response in rostral inputs glutamatergic AMPA-type latency the short receive neurons these to project they as CoPAvia established presumably is interneurons, connection sensory rostral This response. full the triggering for sufficient is cord spinal the of part rostral most the the only that to and cord, confined spinal are responses touch-evoked for required circuits the that suggest experiments Transectionactivation. receptor AMPA-typeon glutamate dependent is response touch the that spontaneous coiling and touch-evoked responses show distinct developmental time courses and these to contribution early motorbehaviorsinimmaturezebrafishisstillunclear.their We demonstratethatthekineticsof but cord, spinal embryonic the throughout distributed Glutamatergicare hours. neurons 30 by infrequent become coils behaviors spontaneous These until coils. contemporaneous contralateral are with touch to responding start they later hours few a and zebrafish embryos show two simple motor behaviors. First, they coil their trunks spontaneously, of genetic studies of circuit formation in this model system. During their first day of development, understanding to the prerequisite development only of not locomotion, is but embryos is zebrafish also of necessary cord for spinal maximizing the the in potential connectivity Characterizing Cellular Biology, GRSNC, CENUM, Université deMontréal, Quebec, Canada 1 T. Pietri Embryos Zebrafish of Cord Spinal the in the Loop DrivesRostral Glutamate Toucha through Response Neurobiology II:Sensoryorgans Institute of Neuroscience, University of Oregon, Eugene, USA;Eugene,Oregon, of University Neuroscience, of Institute 1 , E.Manalo 1 , J.Ryan 2 , L.Saint-Amant 2 andP. Washbourne 2 Department of Pathology and Pathology of Department 1 198 269

Posters Posters 270 C. Norden Interkinetic nuclearmigration inthezebrafish retina:actomyosin forces aretheprimemover again, hereturnstothebarinafastanddirectedmanner. the bar. Between drinks the partygoers jostle around the room, but when one of them gets thirsty of and go to the bar to get a drink. Then they are reminiscent move or are pushed away to make room for others at IKNM during movements nuclear movements of that people at a crowded propose party held in a we room with a bar findings at one end. People our get thirsty on Based is cytoskeleton microtubule the inhibited whenMyosinIIactivityisblocked. when occur still movements However,demolished. or destabilized severely are movement apical rapid as well as stochastic these as microtubules, along We also demonstrate that IKNM is driven largely in by actomyosin contractions mitosis. rather movements than transport nuclear directed, and gradual record being than nuclear to movements rather are that, mainly stochastic show microscopyexcept for and brief cells apical nuclear confocal neuroepithelial translocations retinal preceding zebrafish time-lapse high-resolution use We cell the of phases G2 systematically studied. and G1 the during been never have it back drive that mechanisms the and respectively. and IKNM cycle, of Yet kinetics the basal to apical from migrations interkinetic gradual called involve to believed process generally are IKNM a of movements nuclear undergo The during (IKNM). migration progenitors nuclear expands neural system of nervous nuclei central development, vertebrate embryonic the of epithelium neurogenerative the As Neurobiology, Anatomy, MedicalCollege of Wisconsin, Milwaukee, USA 1 Physiology Development and Neuroscience, University of Cambridge,UK,of Neuroscience, University Physiology and Development 199 1 , S. Young 1 , B.Link 2 , W. Harris 2 Neurobiology II:Sensoryorgans 2 Dept.Biology,Cell suggest thatsensoryfunctioncouldplayaroleinorgan connectivity. fish mutant deaf using results Preliminary innervation. sensory of dynamics long-term the over We are now investigating the contribution of hair cells and neuron activity on target recognition results Our regeneration. hair-cell during provide theanatomicalbasisforsignal-polarityresolutionbylateralline. targets oriented identically re- can with that synapses polarity of establish selectors strict are neurons afferent that show also We forms compartments. neuron Each cells. hair of polarization synapses with planar we hair cells of identical the orientation to divide the recognize neuromast cells, into functional polarised can neurons hair afferent line of innervation appropriate lateral Wethat organ. show lateral-line the in architecture sensory-neuronal of remodeling and the govern that conducted a systematic and comprehensive study of the processes that underlie the elaboration mechanisms the investigate To or alteredphysiologicalconditions. sensory circuitry whose dynamics can be visualized the neuromast can ambiguity.regenerate and be re-innervated very stimulus-polarity rapidly. This provides a a simple model of within creating cells hair neurons, afferent two least at by innervated is neuromast each While stimulus. mechanical other a of component each vectorial the resolve toanimals these how unknown relative hair is it whose Therefore, 180° number, in at equal oriented cells, are hair the of bundles subpopulations line, use two lateral the we of of consist units processes, neuromast, Sensory called system. these model investigate a as To line” survival. “lateral mechanosensory and zebrafish interactions social behaviour, for crucial is innervation sensory-organ of refinement continuous the as well as establishment The Cell andDevelopmental Biology, CRG, Barcelona, Spain A. Faucherre, J.Pujol-Marti,H.López-Schier lateral-line the zebrafish in neurons asstrictselectorsofhaircellpolarity innervation sensory of refinement and Establishment Neurobiology II:Sensoryorgans in vivo over long periods and under normal 200 afferent : 271

Posters Posters 1 272 B.K. Polok Bigh3 isupregulatedinregenerating zebrafish fins The expression of expression The matrix anextracellular as identified protein reportedtoplayaroleincelladhesion,aswellcorneaformationandosteogenesis. first protein encodes kerathoepithelin, called also Bigh3 gene induced TGF-β the target its if (tgfbi). one and signaling TGF-β on focused we Here Wnt. and signaling pathways play a second largest role in that multistep process, like arethe Bmp, Fgf, Notch, retinoic acid, Shh factors Signaling category, beenstudied. regulated during regeneration after regulators of wound healing. Major developmental hasalready regeneration fin Zebrafish to serve can such capacity ofhumans. as and occurs process study this to regenerating the improving on, later and regeneration of process the of whichknowledge our model improve good in a rapidity is the Zebrafish of digits. the because of regeneration tips regenerate only tissue.Mammals, can hand, toregenerate other the theability in have species amphibians urodele and fish Teleost depends onregulationofTGF-βsignaling,suggestinganewroleforthisprotein. of expression of induction the Zeb-IRO, IRO, Sion, Switzlerand; 201 1 , M.F. Bustamante in zebrafish fins was previously reported, here we demonstrate that demonstrate we here reported, previously was fins zebrafish in bigh3 2 , DF. Schorderet correlates with the process of fin regeneration, and also and regeneration, fin of process the with correlates bigh3 2 IRO, Sion, Switzerland 2 Neurobiology II:Sensoryorgans gain-of-function analysisofneuralcircuits andbehavior. and loss-of-function precise for Togethertoolkit ChR2. optogenetic an introduce studies our PLoS in al., sustaining saccades. The saccade deficit in mutants et could be rescued by locally activating (Muto screen forward-genetic unrelated an Genet. 2005), our In group had identified eyes. a sodium-channel mutant, both in full- saccades elicited blown population neuronal this of other activation the in ChR2-mediated Conversely,not but direction. side, stimulated the of direction the in eyes both of saccades blocked NpHRsuppressed of stimulation stimulation Unilateral unaffected. movements Bilateral eye pursuit slow left but completely saccades larvae. zebrafish in (saccades) movements fast of eye generation the for sufficient and required is that VI nucleus cranial the to proximity we used visual behavioral assays (optokinetic response) to identify a cell population in close behavior.swimming Furthermore, studied the throughout hindbrain the of control temporal spinalization” revealed that the central pattern generators “reversible in the spinal cord NpHR-assisted were under tight The (ChR2). channelrhodopsin-2 channel cation light-gated the with activation and NpHR with silencing of combination a by cord spinal and hindbrain in caudal the hindbrain in that control forward cells swimming and localized determined the kinetics we of the technique, swim command this Withbehavior. on silencing their of the test effect simultaneously to and cells in of groups small or target to broadly us enabled head animal’s NpHR the express to submission) data direct specific subpopulations ZFIN.org of neurons. Applying light al., through thin optic fibers positioned et above Mason L. 2007; (eNpHR) Methods Nature al., enhanced et (Scott laboratory our the from lines trap enhancer WeGal4 and NpHR. used original the for lines zebrafish effectively different four across stimulation compared NpHR that showed suppressed spiking of single neurons optically system. clear, recordings theGal4/UAS We they are of oflife. electrophysiological control weeks Loose-patch under since two NpHR first expressing the zebrafish within transgenic foroptogenetics, behaviors generated neurons. many display transfected organisms and of tractable model silencing genetically reversible ideal and are potential Zebrafish membrane for allows of pump, control chloride microbial optical light-driven a (NpHR), halorhodopsin of Expression Physiology, UCSF, SanFrancisco, USA A. B. Arrenberg, P. Schoonheim,F. Del Bene,H.Baier Optical controlofzebrafish behavior withhalorhodopsin Neurobiology II:Sensoryorgans in vivo . Surface localization and spike suppression was didy, with a specific deficit 202 273

Posters Posters 274 cell formation. time receptor taste first for epithelium the pharyngeal the for within required show critically Weis signaling epithelium. Fgf that pharyngeal the within signaling receptor Fgf blocked bud formation. To dissect the role of Fgf pharmacologically signaling with in taste embryos bud cell and type formation, mutant compromised we Fgf receptor specifically function, ace respectively, showed the that Fgf signaling is in necessary for taste function of loss their and epithelium manner in the oropharyngeal epithelium. Fgf8 and the Fgf receptors are expressed firstcharacterized in the pharyngeal we pattern a in located and types type formation, cell distinct of composed are that found and buds taste zebrafish cell bud taste specific underlying mechanisms molecular the the understanding elucidating se. start per To supportive) receptor, in presynaptic, (taste types cell bud taste specific achieved been has progress mechanisms underlying papillae formation, much little is known about the development Although of pharyngeal papillae. the structures, epithelial in harboured be can species the on depending and oropharynx vertebrate the in manner patterned a in located are They cells. support and receptor,presynaptic sensory Taste buds are the vertebrate sensory organs of taste composed of distinct cell types including taste Paris, Superieure, Normale France Ecole U784, INSERM Developpement, du Moleculaire Genetique M. KapsimaliandF. M.Rosa Fgf signalingisnecessaryforpharyngealtastebudreceptorcellformation 203 Neurobiology II:Sensoryorgans lateral lineorgan. proneuromast formation and proneuromast deposition, ensuring the proper segmentation of the primordium. These results reveal that Wnt/β-catenin signaling Wnt/β- coordinates the Interestingly,primordium migration, catenin primordium. activation the domain within does formation not oscillate proneuromast but to remains Fgf lead constant, of irrespective that of expression ligands the stimulates length signaling of the Wnt/β-catenin addition, In signaling. Wnt/β-catenin depends on proneuromasts deposited between spacing and proliferation that show we pathways signaling lineWnt/β-catenin and Fgf the manipulating By primordium. lateral the of region leading the in though Even proliferation cell of rate the understood. by controlled is that event primordium-autonomous a is deposition well not line segmentation occurs by a yet very different mechanism. Our data demonstrates that neuromast as lateral that show other, we each resemble superficially is, segmentation somite and segmentation deposition neuromast periodic not only controls migration but also periodic neuromast deposition. The mechanism controlling signaling Wnt/β-catenin that demonstrate we Here migration. cell directed for important is that primordium the within polarity tissue maintains network interaction genetic This zone. trailing localized on based is network The activation and primordium. of the leading Wnt/β-catenin multicellular pathway in the migrating the leading in the zone and cells restriction of of between Fgf zones signaling to occur trailing the that events signaling cell-cell the described recently Wehave intervals. regular fairly at primordium the of edge trailing the from organsdeposits prosensory sequentially that (primordium) placode migrating a from develops system line The vivo. lateral in patterning and specification mechanisms, fate cell developmental migration, cell fundamental regulation, cell studying stem as for such model excellent an is line lateral The University ofUtah, Dept. and ofNeurobiology Anatomy, SaltLake City, USA T. Piotrowski, A. Aman, R.Crosbie,MT. Nguyen Wnt/b-catenin signalingregulatesmorphogenesisofthelateral line Neurobiology II:Sensoryorgans 204 275

Posters Posters 276 Euan and Hopkins Johns at Research ALS for MacDonald Centre Disease. forMotorNeurone Center Packard Robert by funded is work This lesioned the in thespinalcircuitry thatallowadultzebrafishtosuccessfullyrecoverformalesion. in fibers dopaminergic and serotoninergic spinal cord. We of hope that these studies will increase our understanding of the adaptive density changes the in and interneurones, in the lesioned spinal cord. We are also analysing changes in other cell types, such as inhibitory not contrast, are and In lesion spinal plateaus. a labelled by BrdU. This demonstrates recovery cell after type specific differences in cell death andweeks proliferation functional two number when in post-lesion, change not weeks do cells 6 parvalbuminergic least at to up for level increased this at remained and fish unlesioned to compared post-lesion weeks two at 4.5-fold increased were cells serotoninergic of numbers Overall neurones. motor from distinct are cells serotoninergicthat revealed transferase acetyl choline and serotonin for immunohistochemistry we show by BrdU labeling that serotoninergic cells are also newly born after lesion. Combining into the spinal circuitry after a spinal lesion (Reimer et al., 2008, J. Neurosci. 28, 8510-16). Here that shown lost motor neurones are replaced by newly born motor neurones that mature and are integrated have we Recently, types. cell different adult for specific unlesioned markers in immunohistochemical circuit locomotor the of components zebrafish main and changes the thereof after lesion study and during recovery. to cord aim To spinal this We a aim we after lesion. use weeks transgenic and six within ability locomotor regain zebrafish humans, to contrast In Centre forNeuroregeneration, University ofEdinburgh, UnitedKingdom V. Kuscha, T. Becker, C.G.Becker Neuronal circuit inthespinalcordofzebrafish beforeandafterlesion 205 Neurobiology II:Sensoryorgans ih 10 with challenge, developmental extraordinary an represents brain vertebrate the of wiring proper The Rockefeller The Neuroscience, Sensory of University, Laboratory New York, and USA Institute Medical Hughes Howard A. Nagiel,D. Andor-Ardó, and A. J.Hudspeth the zebrafish of line lateral posterior the in cells hair plane-polarized onto synapses afferent of Specificity Neurobiology II:Sensoryorgans n h cnet f ra rgnrto, u rsls sals te otro ltrlln ogn as organ lateral-line posterior the the for establish system results vertebrate our a regeneration, and organ duringdevelopment of context connectivity the ofneuronal in pattern the in specificity documenting By hair-cell after pattern synaptic innervation vesicle-loaded ablation andregeneration. biased bearing their reassume synapses neurons afferent afferent Moreover, are ribbons. contacts these that level microscopic electron- the at confirm we mCherry, and peroxidase horseradish comprising protein fusion a HRP-mCherry,neurons in expressing By spatially consistent. are is preference organs polarity innervated the and the consecutive but neuromasts, multiple on contacts form neurons the of specificity and find evidence for bias from as early as 2.5 days post-fertilization. More than half hair- same the of this hair-cellpolarity,quantify of cells we selector biased a as hair neuron polarity.the bundle Modeling with contacts specific establishes neuron afferent each that show promoter, we neuronal a of posterior control the under the mCherry protein of fluorescent cells organized the with neurons line lateral hair afferent single labeling By of polarized neuromasts. as known consists clusters discrete zebrafish in larval of organ lateral-line posterior The how synapticspecificityarises. understanding for models necessary provide systems complexity,vertebrate this simple of view In connections. neuronal appropriate of specification the is process this of feature understood in governingsynapticspecificityduringtheformationofneuralcircuits. mechanisms -independent and activity-dependent of role the of analysis experimental detailed 11 ern frig n siae 10 estimated an forming neurons td o snpi tre slcin Ti sse pris a permits system This selection. target synaptic of study vivo in 15 yass n uas A esnil u poorly but essential An humans. in synapses 206 277

Posters Posters stimuli were found to evoke motor behaviors in both wild type siblings and siblings type wild both in behaviors motor evoke to found were stimuli These (pinch). harsh-touch and isothiocyanate) (allyl oil mustard including stimuli noxious and neuron function neuron frommechanosensitive feedback in torequire neurons trigeminal known response a behavioral head-first, objects ) complemented the complemented touchdown) and (macho mutants touch-unresponsive identified previously with crossings pair-wise Finally 278 2 Low S. touché, anewtouch-unresponsive zebrafish mutant mechanosensitive neuronsforresponsivenesstolight-touch. 1 during the first two days of development. One family, subsequently named families were screened for progeny exhibiting reduced or absent touch-evoked motor behaviors question we undertook a chemical based (ENU) forward mutagenesis within mechanically sensitive screen neurons signal is a largely electrical unresolved in question in neurobiology.an zebrafish. into Totransformed Mutated address is this vertebrates in light-touch which by process The Arbor, USA Amant that Further examination revealed that offspring. touch-unresponsive completely in results which mutation recessive a harbor to found eatet f oeua, ellr n Dvlpetl ilg Uiest o Mcia, Ann Michigan, of University Biology Developmental and Cellular Molecular, of Department CellularPathologyBiology,and of Department Montréal,de Université Montréal, QC, Canada; 207 tou 1 1 J Ryan J. , mi173 represents a new touch-unresponsive mutant, and suggest that suggest and mutant, touch-unresponsive new a represents 1 tou SM Sprague S.M. , mi173 Xenopus laevis Xenopus mutants were exposed to stimuli which activate the lateral-line system, lateral-line the activate which stimuli to exposed were mutants tou 2 H Hirata H. , tou mi173 mi173 embryos. To rule out a wholesale disruption of sensory of disruption wholesale a out Torule embryos. mutant phenotype. Collectively these findings indicate findings these Collectively phenotype. mutant mutants also fail to stop swimming after encountering 2,3 W Zhou W. , Neurobiology II:Sensoryorgans 2 W.W.Cui , 2 JY Kuwada J.Y. , touché ( is required by required is tou tou mi173 tou 2 L Saint- L. , mi173 mutants. ), was population ofglutamatergic synapsesinanintactvertebrate. sensory-motor behavior. This is the first description of protein recruitment to a precisely defined a reveals recruitment synaptic of a for critical is that synapse line a at components postsynaptic and pre- both of assembly sequential time This 19hpf. at seen first were PSD-95, SAP97 proteins and the PSD-93, for immunoreactivity with side, contacts postsynaptic at the localized On hpf. cytoskeleton, 21 the after to cells CoPAvesicles synaptic tether that proteins 2, synapsins and contrast, 1 In VAMP2. tagged fluorescently this of and imaging examined by points confirmed time was all distribution at axons RB throughout distributed equally by were SV2 with components synaptic during straddles of the time of initiation form of the touch recruitment response at 21hpf. interneurons Synaptic vesicle precursors of labeled CoPAsynaptic labeling between 16 to 25 hours and timeline post fertilization (hpf), a developmental timeframe that cells the RB immunolabeling and characterized live imaging between techniques using pre- we and synapses postsynaptic markers. development, We the examined how understand To through (CoPAs) interneurons commissural to on signal glutamatergic synapses. the pass and stimulus the transduce in consists response, touch an the behavior, underlies coiling of This that the trunk circuitof embryo. the embryo a zebrafish in response of the to a part tactile in stimulus. Rohon-Beard forms behavior sensory neurons that motor synapse early a These at vivo components. in issue necessary this We re-examining packets. all transport are precursor synaptic of as together arrival trafficked be to concomitant thought are the components by formed are specializations Studies of synaptogenesis in primary neuronal cultures have led to the hypothesis that presynaptic Institute ofNeuroscience, University ofOregon, Eugene, USA P. Washbourne, C.Easley-Neal,andT. Pietri Synapse Synaptogenesis Neurobiology II:Sensoryorgans : Sequential Recruitment of Components to a Physiologically Relevant Physiologically a to Components of Recruitment Sequential vivo: in 208 279

Posters Posters 280 vision. paralogues. In the future, knockdown experiments will determine the function of these genes in mGluR6 the of localization synaptic the identify to antibodies generating of process the in are the that likely it Since pathway. signalling mGluR6 the by coexpression with supported was result surprising This cells. ganglion retinal in predominantly genes both located we cells, bipolar in expressed is mGluR6 where mammals, to contrast In paralogues. mGluR6 two the on focused we retina, mammalian the in function specific the to Due paralogues. gene paralogues, for two example between for patterns expression found Different as retina. the in mGluRs different the of all for pattern expression unique RNA event. duplication genome we whole 8) specific and teleost 6, a in 5, arose likely 2, Those paralogues. (mGluR1, two ortholog members mGluR mammalian each most for detected For mice. and mGluR the humans of to phylogeny comparable the is subgroups, family three into division its With Conclusion: and Results adult zebrafishretina. and developing the in pattern expression respective their determine and family this of members all clone to out set we retina, teleost the in mGluRs of role functional the elucidate to step first a As retina. mammalian the in response ON the mediate to found is and dendrites cell bipolar ON on expressed is which mGluR6, is transmission synaptic direct in functioning mGluR an of example only The release. neurotransmitter regulate likely they where retina, vertebrate the synapses of all at identified been have (mGluRs) receptors glutamate Metabotropic Background: Institute ofZoology, University ofZurich, Switzerland M. Haug, Y-Y. Huang,M.Gesemann,S.C.F. Neuhauss Metabotropic glutamatereceptorsinthezebrafish retina 209 paralogues are expressed below detection level in bipolar cells. We cells. bipolar in level detection below expressed are paralogues mGluR6 gnao (coding for Goα proteins), the G-protein which is thought to conduct and mGluR1a, in 5 day old zebrafish, supporting the distinct roles distinct the supporting zebrafish, old day 5 in mGluRs is also expressed in inner retinal cells, we deem we cells, retinal inner in expressed also is gnao sget dsic sbucinlsto o both of subfunctionalisation distinct a suggest -1b, yrdzto eprmns eeld a revealed experiments hybridization situ in Neurobiology II:Sensoryorgans of thesedefects. hair cells in the inner ear. three We are currently mapping these loci to discover the underlying cause and pit show nasal the in cells olfactory we including neurons sensory other of Here, ciliogenesis the in affect dpf. mutations these 5 that found We photoreceptors. at in development ciliary development affect which mutations photoreceptor affect isexpressed which GFP mutations line, several this In zebrafish. of line transgenicspecifically in the rod photoreceptor cells under the control of rhodopsin promoter. GFP We isolated a using screen pressure earl To identify mutations that affect the photoreceptor development and maintenance, has we performed photoreceptors been poorlyunderstood. in maintenance and formation ciliary of mechanism molecular However, proteins causes photoreceptor cell death and progressive blindness called as retinitis pigmentosa. cell body to the outer segments thorough cilia. the In human, functional disruption of and several ciliary segment large outer amounts of proteins involved the in photo-transduction are transported from between the photoreceptor connects axoneme photoreceptor cell microtubule body.cell Ciliary a the integrity is of important with for survival side of photoreceptor apical cilium cells, because the A on body. segment outer photo-sensitive a develops cell photoreceptor A Y. Omori A gfp-basedgeneticscreenreveals mutationsaffectciliogenesisinphotoreceptorcells Neurobiology II:Sensoryorgans 1 Osaka BioscienceInstitute, Osaka, Japan; 1 , J.Malicki 2 , T. Furukawa 1 2 Harvard MedicalSchool,Harvard Boston, USA 210 281

Posters Posters and itsmappingisunderway. to this mutant does not rescue the muscular defects. The mutation is located in chromosome 21 in endoderm type wild of Introduction cells. crest neural and muscular epithelium, pharyngeal defects provoking the OL185 phenotype, we are currently performing rescue experiments of the pharyngeal wall) are severely reduced or missing in OL185 mutants. To give insight to the tissue In mutants. altered OL185 conformation. In addition, the branchial in muscles (transverse ventral 1 to 5 altered and the dorsal severely are archesparticular, the second pharyngeal arch pharyngeal and the five branchial arches are reduced and present an the of structures cartilaginous that the mutation is not involved in notion the formation of the taste bud tissues supporting of origin. However,mutant OL185 ventral the in affected not is markers reduced epithelium oro-pharyngeal a have mutants OL185 that late and early of expression the Second, found distribution. regional altered and buds taste of number we First, oropharynx. surrounding the and types cell bud taste the for specific markers analysed Towe defects. phenotype, cell this characterise receptor taste on based isolated, was mutant OL185 the screen, mutagenesis ENU an Through forward geneticapproachinzebrafish. of taste bud formation remain largely unclear. To address this question, we are using primarily a aspects cellular and molecular are The arches. pharyngeal They palate, lips, the development. on patterned regionally of from day mainly second arise the after buds epithelium taste pharyngeal the endoderm-derived zebrafish, the In cells. glial-like the and organs, presynaptic receptor, sensory taste taste the of composed organs, by multi-cellular onion-shaped, are tastants buds taste differentiated different The buds. taste detects gustatory system the this system, chemosensory vertebrates, a In on system. mainly relies alimentation proper of selection The Germany 282 G. Gibon The zebrafish OL185mutationaffectspharyngealandtastebuddevelopment 1 INSERM U784 - Ecole Normale Supérieure, France; 211 1 , Tübingen2006ScreenConsortium 2 , F.M. Rosa 2 Max-Plack-Institute für Entwicklungsbiologie, Neurobiology II:Sensoryorgans 1 andM.Kapsimali 1 12 cells at 48 h after transfection. In contrast, only PC-12 cells transfected with pCMV-zEfn- with transfected cells PC-12 only contrast, B1 In transfection. after h 48 at cells 12 PC- in outgrowth neurite significant induce could proteins zEfn-B-GFP all with that NGF,found treated we and cells PC-12 into transfected were constructs expression pCMV-zEfn-B When four zebrafishephrin-Bproteinsenhancesneuriteoutgrowthinembryos. zEfn-B3 is inhibitory to the neurite extension in vivo. On the other hand, and the zEfn-B1 forward of signal signal of reverse all the that suggesting neurons, GFP-labeled from outgrowth neurite vivo andinvitro. in outgrowth neurite of regulation differential has signal reverse ephrin-B Altogether,zebrafish signal of zEfn-B1 and zEfn-B3 has the potential to promote the neurite extension in PC-12 cells. C-J. Huang in ephrin-b3 zebrafish embryos andinpc-12cells and ephrin-b1 of signal reverse by outgrowth neurite of regulation Differential Neurobiology II:Sensoryorgans n epesd hm ne te ae u pooe floe b te neto it zebrafish into injection the by followed promoter embryos at 1-cell stage. Interestingly, HuC zEfn-B1 same the under them expressed protein and ephrin-B zebrafish each of domain intracellular the deleted further We not. could HA zEfn-B3- and zEfn-B1-HA but neurons, GFP-labeled from outgrowth neurite significant induce could zEfn-B2b-HA and zEfn-B2a-HA only that found we stage, one-cell at embryos zebrafish expressionof drive the to HuCpromoter each Efn-B with the HA-tag. When pHuC-GFP was co-injected theneuron-specific with each pHuC-zEfn-B-HA into we used outgrowth neurite embryos, regulate zebrafish to member in ephrin-B each and of ability B2b the B2a, investigate the ephrin-B1, bidirectional to through order zebrafish, In in signal B3. mediate members reverse (Efn-B) a ephrin-B could and four are receptors There which Eph itself. the ligand through molecules, signal forward transmembrane a with are signaling, proteins ephrin-B The Sinica, Taipei, Taiwan 1 Institute of Biological Chemistry and Chemistry Biological of Institute Δ C- HA and pCMV-zEfn-B3 1,2 , G-D.Chen 1,* , K-L.Huang Δ C-HA did not show neurite outgrowth, suggesting that the reverse 2 Institute of Cellular and Organismic Biology,Academia Organismic and Cellular of Institute 1,* , C-H.Cheng Δ C-HA and zEfn-B3 1 , B-K.Wu Δ C-HA could induce significant 1 , C-C.Hung 212 1 283

Posters Posters 284 processes ofthesensorycellsinlateralline. functional significance of theseThe resultsTRKB. remainsexpress stillto obscurecontinue butthey italthough couldindependent bematurity,become relatedreach toneuromast regenerative when nevertheless, manner; paracrine/autocrine a in acting neuromasts involved of is development the system in factor growth subpopulation TRKB/BDNF small the that a suggest to results was Present restricted cells. it being sensory andof animals older development, of with neuromast decreased from progressively absent neuromasts, all was observed dpfalmost 4 at immunostaining cells BDNF hair all specific in but BDNF Faint ganglia. line changes. lateral posterior developmental and undergoes anterior was TrkBthe moreover in neurons examined; of time subpopulation the a in through found neuromasts of cells sensory the in detected sqet4) from 4 days postfertilization (dpf) to 120dpf. Specific TrkB immunoreactivity was regularly TrkB/BDNF system in neuromast of wild type and transgenic zebrafish (strains ET4, Et (krt4:GFP) never investigated. In this study we used immunohistochemistry to investigate the expression of were neuromasts zebrafish in TRKB/BDNF of expression the the in changes know related including developmental we as species, far As fish zebrafish. the different in in development embryonic levels throughout BDNF protein and TRKB or mRNA the both at detected express been have Fishes which respectively. neuromasts, trunk-tail and (BDNF,common (TRK), NT-3, receptors head (NT-6, NT-4) their specific NT-7) and or neurotrophins the innervate which the to ear,behind or anterior located ganglia, cranial two of peripheral neurons the sensory by bipolar of supplied processes are and cells, mantle and supporting hair, sensory of consist They The neuromasts are the structural units of the lateral line system in bony and cartilaginous fishes. Oviedo, Spain Celular,de Biología Universidad y Morfología de *Departamento Italia;Messina, di Studi degli e CISS (centro di Ittiologia sperimentale per la Sicilia), Facoltà di Medicina Veterinaria, Università Dipartimento di Morfologia, Biochimica, Fisiologia e Produzione Animale, Sezione di Morfologia, J.A. Vega*, A. Germanà,R.Laurà,G.Montalbano,andV. Amato TRKB/BDNF expressioninthehaircellsofdeveloping lateral linesysteminzebrafish 213 Neurobiology II:Sensoryorgans Although we have detected BrdU detected have we Although was alsocarriedout. Islet1. and double immunohistochemistry for BrdU and neural progenitor markers like Nestin, Prox1, Pax6 developed we RPCs the in markers retinal of pattern expression the of analysis the For week. a the in RPCs of detecting the during solution BrdU-containing a of For injection intraperitoneal out carried have we retina, life. adult adult throughout continuous rods have new animals generate these and as growth RPCs, retinal of study the for model excellent an is potential Teleostretina proliferative their studying and . vitro retina the its through during differentiation pattern expression and the describing migration RPCs, the of largely characterization the are is purpose underlying Our mechanisms the although retina, unclear. degenerating a and migrate to within cells these of differentiate capacity the to related directly is RPCs of stage ontogenetic The as function, visual the improves these cellsdifferentiateintomaturerodsandintegratewithintheretina(MacLarenetal.,2006). retina degenerating a into promising RPCs a as of Transplantationrevealed been strategy. has (RPCs) cells progenitor rod of use The lost. already are that cells the replace to or rods and cones of death the delay to either attempting made been studies have numerous so humans, in blindness of cause common a is degeneration Photoreceptor Cell biologyandPathology, INCYL, Salamanca, Spain R. Sánchez-González, A. Santos-Ledo,M.Ogueta, A. Porteros,J. Aijón andR. Arévalo In vitroandex vivo modelsofgeneration ofrodprogenitorcells Neurobiology II:Sensoryorgans Islet1 oeta: tm el, irtn poeios n rd rcros Wees h mgaig RPCs migrating the Whereas (Nestin markers studied the precursors. all almost to positive are rod and progenitors migrating cells, stem potential: proliferative and retina the within location their to according defined be can RPCs of subtypes Three (ONL). layer nuclear outer and (OPL) layer plexiform outer (INL), layer nuclear inner the Barruso, deCastillayLeón Junta andRed Terapia Celular deCastillayLeón. Supported by grants from the Fundación Mutua Médica Madrileña, Fundación Samuel Solórzano to NIHguidelines. for the use care of animals in research (RD the 1201/2005, of BOE European guidelines 252/34367-91, Communities Directives (86/609/EEC the 2005) and 2003/65/EC), with and the current accordance Spanish conformed legislation in were protocols experimental and procedures the All maintains rate proliferative The culture. in stable aftertwomonthsinculture. the days 5-7 by after confirmed neurospheres potential, primary proliferative of great formation a showed cells using These cells progenitor medium. rod proliferative grown a and isolated have we assay, characterization vitro in our In restricted expressionpattern. + ), the stem cells (Pax6 cells stem the ), hybridization against Notch1 and NeuroD after BrdU immunohistochemistry BrdU after NeuroD and Notch1 against hybridization situ In + ) and post-mitotic rod precursors (Notch1 precursors rod post-mitotic and ) + cells in all retinal layers, RPCs are exclusively located in located exclusively are RPCs layers, retinal all in cells + , Prox1 , + , Notch1 , 214 + + , CD133 , , NeuroD , + , NeuroD , + ) have a more a have ) + and 285 in

Posters Posters 286 developed, toaskquestionsabouttheroleofactivityinformationspinalcord. manifestation of early embryonic behaviours and use that information, and the genetic tools we As more data are gathered, we will be able to learn which channel subunits are necessary for the subunit ofthisgene(CaV1.3a)tocheckforpossibleredundancy. be may be to motoneurons, appears normal. We and embryos are currently performing double knockdowns using a morpholino to the duplicated knockdown interneurons CaV1.3b of V2 behaviour as circuitry, in motor for stages dispensable early at expressed although Cav1.3b, targeted the of role behavioural the for Quantitative assay an expression. proteins. as used protein was behaviours perturb embryonic three to the of order analysis in subunits channel these calcium of translation block specifically to designed were morpholinos Thereafter development. Weemployed to ability the develops embryo the hpf, swim, withlowamplitudeandhighfrequencycontractions. 28 around Lastly, the hpf. over 21 around decreases stimulation, gradually tactile and hpf, contralateral 19 to response a as coiling of around onset the is behaviour second peaks The hours. 10 following coiling this of frequency The (hpf). post-fertilization hours 17 around starting coiling, spontaneous of apparition the of consists first motor behaviours to channels in calcium zebrafish.voltage-gated Earlyof motorrole circuitryinvestigate to resultsapproach ingenetic threereverse characteristicwe a behaviours.question, employed The this into insight Tobiology.gain in question central a is behaviour underlie that circuits of function and formation the to contribute encode, they proteins the and genes, How depathologie etbiologiecellulaire,Département Université deMontréal, Canada zebrafish of circuitJ. Ryan , M.Lachance,L.Saint-Amant motor the within embryos channels calcium voltage-gated of Characterization 215 hybridization to assess the expression of several calcium channels during channels calcium several of expression the assess to hybridization situ in Neurobiology II:Sensoryorgans monitor myelination and tight junction formation in vivo and can then serve as a read out for out screens affectingmyelinationandpropertightjunctionformation. read a as serve then can and vivo in formation junction tight and myelination to monitor us allow will lines These junctions. tight autotypic to localization proper their verifying linewith are atransgenic aregenerating we membrane bound eGFP. Currently, Additionally, glia. express we express eGFP myelinating to / mCherry-Claudin k used fusion in proteins and be specifically can it proteins reporters, fluorescent optional and constructs responder and driver independent with 2009) flexible al., the et Using (Paquet PNS. system the Tol2-Gal4-UAS in as claudin k promoter, that drives strong specific expression in myelinating glia therole – in the into CNSa ascloned have we general, in well biology myelin and function k Claudin dissect further to tool a As give insight may which partners, interaction Claudins playinmyelin. find to k Claudin for experiments co-immunoprecipitation performing are we addition, In larvae. zebrafish in experiments down knock performing are and Claudins of relationship phylogenetic the analyzed we question, this Todivergentfunctions? evolved they have or redundant teleosts address in Claudins myelin are 11 and19,allofwhichwefindtobeexpressedinbothCNSPNS.Thatraisesthequestion: restricted to the CNS and PNS respectively, outer zebrafish possess notand only Claudininner k butthe also Claudin and mesaxons – similar to loops, mammalian Claudin 11paranodal and 19. While the mammalian Claudin 11it incisures, and that 19 Schmidt-Lantermann are show the We to oligodendrocytes. localizes and cells Schwann in exclusively expressed is which 31), We have identified a teleost specific, novel myelin protein, zebrafish Claudin k(formerly Claudin fence functionofClaudinsmaycontributetocompartmentalizationthemyelinmembrane. insulation, presumably by regulating ion flow between interstitium and intramyelinic space. The oligodendrocytes (Claudin11/OSP) andSchwanncells(Claudin19)contributetoelectrical domains in epithelia. In myelin of rodents, Claudins are expressed in autotypic tight junctions of across paracellular pathways and function as a fence between apical and basolateral membrane Myelinating glial cells have evolved specific molecular mechanisms and components to and components mechanisms molecular Claudins are essential components of tight junctions, which form selective permeability family.protein Claudin specific the of members include These evolved sheath. myelin their maintain and have form cells glial Myelinating cost. axon diameters keeping small, while permitting conduction compact impulse nerve powerful saltatory nervous rapid, systems allows at This relatively axons. small around metabolic processes cytoplasmic their wrap tightly (PNS) system nervous peripheral the in cells Schwann and (CNS) Myelination is a key step in vertebrate evolution: oligodendrocytes in the central nervous system Biochemistry, Adolf-Butenandt, Muenchen, Germany K. Schaefer andC.Broesamle Myelin-specific Claudinsinzebrafish: evolutionary andfunctional characterization Neurobiology II:Sensoryorgans 216

barriers 287

Posters Posters 1 288 C.M. Maurer Impaired EnergyMetabolismLeadstoReduced Vision intheZebrafish noirMutant n ocuin a uain n mtblc e ezm las o eue vso ad retinal and vision reduced to a complexpathogenesis. leads enzyme key metabolic a zebrafish the in dystrophy in mutation a conclusion, In cells, whichmostlikelyinvolvesdeficientgenerationoftheneurotransmitteracetylcholine. were phenotype We found that the defect in the inner histological retina is due to a selective damage to cholinergic and amacrine ERG diminished acetyl-CoA levelscausedbytheblocklinkingglycolysisandKrebscycle. the and longer significantly improved. The survived long-chain fatty acids provide an alternative larvae energy source the to restore rescue Mutant could acids phenotype. fatty long-chain of morecomplex consisting a diet ketogenic suggesting a Strikingly, mutation the by first energy- affected pathogenesis thansimpleenergy most deprivation. are the retina – the photoreceptors of than cells rather consuming layer, nuclear inner the of cells Interestingly, defect extendstotheouterretinaat7dpf. At 6 dpf defects in the inner dpf. nuclear 7 layer at are b-wave observed in and histological a- sections both, of of the lack retina. complete This a to and dpf 5 at amplitude b-wave ERG the of decrease a to leads activity enzyme of Absence phenotype. mutant noir the underlies complex Genetic mapping of the mutation revealed that a days postfertilization(dpf). defect in a subunit of 5 at starting neurons secondary the to primary from transduction signal of block a Pyruvate suggested which Dehydrogenase (ERG) electroretinography by confirmed was vision behavior.Reduced optokinetic of absence mutant the screen a such In genes involvedinthefunctionofvisualsystem. identify to rerio) (Danio zebrafish the in performed been have screens mutagenesis Large-scale Centre (CNIO), CancerCell BiologyProgram, Madrid, Spain nttt o Zooy Uiest o Zrc, Switzerland; Zürich, of University Zoology, of Institute 217 1 , H.B.Schönthaler mutant. The defect initiates in the inner nuclear layer suggesting layer nuclear inner the in initiates defect The mutant. noir has been identified due to its dark external appearance and appearance external dark its to due identified been has noir 2 , S.C.F. Neuhauss 1 Neurobiology II:Sensoryorgans 2 pns Ntoa Cne Research Cancer National Spanish mutant noir loss ofvisualfunction. markers photoreceptor with retina outer the of staining contrast, In mutants. raf inner retina suggests that amacrine and ganglion cell differentiation is equivalent in siblings and blindness. Preliminary results suggest that specific defects in Conclusions: We have identified and partially characterised a novel zebrafish model of inherited defect inrafphotoreceptorsandretinalpigmentepithelium(RPE). morphological a suggests data microscopy light Preliminary normal. is retina peripheral the in appears tobereducedoverallinrafmutants.However, stainingofnewbornrodphotoreceptors ee eemnd y UE sann. h itgiy f h inr eia haod ad trunk novel A and Results: (hyaloid) System). retina Zebrafish (ViewpointTracking Zebralab using inner ascertained were behaviours the response of integrity The Circadianshadow- staining. and phosphatase locomotion alkaline by analysed staining.was vasculature TUNEL by determined were or levels of cell death is observed between mutants and siblings. Light microscopy and DAPI and show microscopy Light staining siblings. and mutants between observed is death cell of levels morphology or vasculature in difference No reponse. shadow a and behaviour circadian exhibit show a significantly reduced or absent OKR and are paler with slightly smaller eyes.The smaller withslightly arepaler and OKR of levels activity or absent locomotor reduced significantly a show mutagenised zebrafish to identify families with recessive defects in visual function. Retinal markers cell-specific the visual using sections retinal in of defects with recessive differentiation and families lamination was identify characterised by to light microscopy, zebrafish and immunohistochemistry mutagenised Methods: Optokinetic response (OKR) screens were performed on N-ethyl-N-nitrosourea (ENU)- Purpose: To characterisethebasisofinheritedvisualfunctiondefectinRaifteirímutant. UCD SchoolofBiomolecularandBiomedicalSciences, University College Dublin, Ireland B.N. Kennedy, A.L. Reynolds,B.Sapetto-Rebow, K.J.Curtin,L.Shine,D.Cottell, Y. Alvarez Defective photoreceptorsunderlieinheritedblindnessintheraifteiri mutant Neurobiology II:Sensoryorgans uat t hv nra rtnl lamination. retinal normal have to mutants raf mutant designated mutant uat ae 5% f oml ilns however, siblings, normal of ~50% are mutants raf Raifteirí (raf) Raifteirí zpr1, was identified by OKR screens. OKR by identified was zpr3, raf outer retina and 5e11 and 5e11 218 . Cell death levels death Cell zn-5. morphology underlie tiig f the of staining zn-5 and zpr3 mutants Raf mutants raf zpr1 289

Posters Posters mutations in for screening are we addition, In imaging. vivo in by and tissue fixed in polarity both stages, different at photoreceptor of development the of analysis detailed a performing currently are We in photoreceptorandkidneyepithelialcells. morpholino- induced zebrafish, loss In of cells. photoreceptor vertebrate of segment inner the to corresponds crumbs of absence Drosophila, In neuroepithelium. leads to the a reduction in of the length structure of the polarizedstalk membrane, a the portion of affect the apical okomembrane which 290 of apico-basalpolarityphotoreceptorcells. the cell biological function of the Crumbs protein complex in the differentiation and maintenance the of one in Mutations and the scaffolding proteins Stardust, epithelial and photoreceptor cells. The core components are the transmembrane protein Crumbs the Crumbs protein complex has been shown to is control the which differentiation surface, of the apical apical surface their in subdivided into in the outer segment, the connecting cilium and the inner segment. In evident particularly is cells photoreceptor mature in Polarity function. specialized highly their for morphology,essential unique a have cells Photoreceptor Max PlanckInstituteofMolecularCell BiologyandGenetics, Dresden, Germany M. LuzandE.Knust Differentiation andmaintenanceofzebrafish photoreceptorpolarity The zebrafish genome encodes five encodes genome zebrafish The dependent retinaldegeneration. to disintegration of embryonic epithelia, morphogenetic defects in photoreceptor cells and light- 219 crb crb2b function has been shown to affect the differentiation of apical characteristics genes by TILLING. We will report on the results, which can provide insights on orthologues, crumbs genes, a single a genes, crumbs DPATJ and , and in the in and okomeduzy, DLin-7. Defects in individual components lead Neurobiology II:Sensoryorgans gene and three and gene stardust stardust stardust orthologue , Drosophila genes. lin7 nagie n ocuin or aa eosrt ta cns s a iia mcaim o os o fast for rods to mechanism similar a use cones that demonstrate photopigment inactivationandforensuringahightemporalresolution. data our functional conclusion, In elucidate to visual behaviortestingrevealedthatthisdelayimpairsperformance. order subsequent in and recovery.photopigment in delay a cause levels protein arrestin reduced that Moreover,showed arrestins performed inactivation. photopigment the the were to arrestins cone of of contributions recordings down (ERG) knock electroretinographic Morpholino-mediated analysis. protein and by mRNA expression specificity type cell their verified and arrestins cone two identified We arrestin onquenchingconesignaltransduction. of impact the study particular,to in and, photoreceptors, cone in inactivation photopigment of Here, we used the cone-dominated retina of zebrafish larvae toone in rods. geneticallythe dissectto similar themechanism mechanism quenching a suggesting - inactivation response cone proper for Recent studies have shown that phosphorylation of the photopigment opsin is also a to crucial step protein arrestin the of phosphorylated rhodopsinisessentialforthecompleteinactivationphotopigment. binding Subsequent rhodopsin. activated of phosphorylation the by dim light (scotopic) conditions. The initial step in shutting-off rod signal In transduction contrast, is rod comprised photoreceptors are highly sensitive to and light cones and, photobleaching therefore, avoiding responsible ensuring hightemporalresolution. response, for are cone vision the at of course only time the shaping Not mechanisms the function. photoreceptor about known However,is vision). little (photopic levels light ambient high cone of presence in even by mainly governed responsible for color vision; they also enable the retina to rapidly signal changes in illumination is vision Human Institute ofZoology, University ofZurich, Switzerland S.L. Renninger,M.Gesemann,S.C.Neuhauss Arrestin availability inconephotoreceptorsmodulatesopsininactivation Neurobiology II:Sensoryorgans Paired-flash ERG recordings ERG Paired-flash 220 291

Posters Posters 292 rather thanhairbundlemaintenanceorcelldifferentiationperse. that propose together,Takingwe results assays. these and type wild in between found adecrease shows phalloidin were differences However,statistical patches. coupled no sensory distinct in structures these fluorescein of number morphological using addition, structures In bundle syndrome. hair of human analysis the of aspects phenocopying thus when swimming, defects balance presented respond, did that those that assay,but hearing tapping the to 5dpf the of most that show results Our development. patch knock-down approach, we are currently characterizing the function of a morpholino and a model as zebrafish the Using pigmentosa. retinitis and areflexia vestibular on mutations humans, In contain isoform. that translated the proteins of the function scaffold in domains cytoskeleton binding PDZ conserved 3 or evolutionary 2 for codes gene USH1C deflection ofthehairbundleandonitscorrectintegrity. brain. Thus, the transformation of the mechanical input into a chemical response depends on the generate an action potential that will be driven towards the different interpretation centers in the hair cell and released into the synaptic cleft of the innervating neurons. Responding neurons will the apical side of the hair cell. As a result of the deflection, chemical signals are produced by the interconnectedan staircasea in arranged structure microvilli in actin-rich localized and pattern auditory and ear,vestibular inner the stimulation In cells. hair will the balance: physically and hearing of induce functions the a for essential vectorial are that cells deflection of the hair bundle, The inner ear is an organ characteristic of the vertebrates. It rooms specialized mechanosensory Institute ofNeuroscience, University ofOregon, Eugene, USA B. Blanco,J.B.Phillips,Wegner andM.Westerfield The roleofush1cduringzebrafish oticsensorypatch development 221 gene cause Usher syndrome type 1 that is characterized by congenital deafness, congenital by characterized is that 1 type syndrome Usher cause gene USH1C morphant larvae in the cell proliferation and cell death cell and proliferation cell the in larvae morphant ush1c Neurobiology II:Sensoryorgans is required for hair-bundlegenesis for required is ush1c morphants did not respond not did morphants ush1c ush1c during ear sensory University of California, San Francisco, USA.;Francisco, San California, of University 1 D. Li Investigating theroleofglialcellsinspinalcordusingchemical optogenetics Behaviour of California in Berkeley,in Berkeley,California USA;of currently investigating in vivo the intra-cellular mechanisms by which depolarization of glial of depolarization cells canmodulateneuronalactivity.which by mechanisms intra-cellular the vivo in investigating are We currently light. the of patterning spatial the on depending motion struggling large a or response escape touch a swim, forward a to correspond either could observed motion The pattern. light glial cells in the spinal cord could induce many different types of locomotion depending on the cells. We conducted light pulse experiments on these lines and found that optical stimulation of in order to select for lines with explicit LiGluR expression in the spinal cord, specifically in glial cerebrospinal fluidneuronsledtoslowswimmingonly. We performedanenhancertrapscreen behaviors: Rohon Beard neuron activation induced an escape like response, while activation of are zebrafish photoswitch previously the that chemical optical stimulation of specific a neurons in in the spinal to cord led (LiGluR) to distinct coupled locomotor receptor channels sensitive to specific light wavelengths and allow UV light to depolarize specific cells. engineered We found glutamate genetically ionotropic These cells. light-gated larval of introduction the on based approach stimulation optical Wean unknown. took still are mechanisms the nervous but system, central the in activity neuronal modulate can cells glial that evidence recent is There Lawrence Berkeley NationalLaboratory, Berkeley, USA nvriä, uih Germany; Munich, Universität, Helen Wills Neuroscience Institute and Department of Molecular and Cell Biology,Cell University and Molecular of Department and Institute Neuroscience Wills Helen 1 , C.Wyart 1* , F. DelBene 2 , E.Warp 4 hscl isine iiin n Mtra Sine Division, Science Material and Division Bioscience Physical 1 , E.K.Scott 2 Department of Physiology, Program in Neuroscience,Physiology, in of Program Department 3 Department of Chemistry, Ludwig Maximilians- LudwigChemistry, of Department 2 , D.Trauner 3 , H.Baier 222 2 , E.Y. Isacoff 1,4 . 293

Posters Posters 294 M. Dadda zebrafish adult in personality and lateralization both influences asymmetry epithalamic left-right Early 1 that otherfactors,i.e.hormonal,maybeimplicatedintheirdevelopment. suggesting with thus sex, wereuncorrelated by influenced strongly while asymmetries social distance functional and anatomical of both Only measures position. parapineal by was turn influenced in that inspection predator of laterality with correlated but asymmetry anatomical of covered a shorter distance when released in the dark. Activity in the open field was independent a shorter distance from predators, spent more time in the peripheral portion of an open field and predator, open field anda socialfrom distance).(distance Fishtraits withpersonality atypicalof rightmeasures parapinealthe position,of maintained some in response different a showed also turns in the dark). Fish with opposite parapineal positions differed in all laterality measures. They swimming and preference rotational (mirror,inspection, adults predator as lateralization motor right parapineal at birth using a genesis of individual differences in coping styles and personality. We sorted zebrafish with left or play a key role in the development of lateralization of cerebral functions and are they involved in that the posited been has it and zebrafish of development the in early very appears structures avoidance learning, and hormonal mating, response feeding, perception, to olfactory as stress such functions and important many reward. in implicated Asymmetry are and in these epithalamic vertebrates all of structures hindbrain and midbrain with areas forebrain limbic the connecting The habenulae and associated parapineal organ are part of a highly conserved conduction system University ofPadova, Italy eatet f eea Pyhlg, nvriy f aoa Italy; Padova, of University Psychology, General of Department 223 1 , A. Domenichini 2 , L.Piffer foxD3:GFP marker and subjected them to four tests of sensory and 1 , F. Argenton 2 , A. Bisazza 1 2 eatet f Biology, of Department Behaviour juveniles willhelpdefiningdevelopmentalaspectsofthephysiologytasteinvertebrates. zebrafish of neurons sensorial the in imaging calcium on based data, functional with combined in functional already are cycloheximide prevent buds feeding taste in larva, or denatonium that like consistent compounds bitter with noticed since food the take to have work begin larvae when of we zebrafish 5day-old others. assay, This this behavioural assay Using time. given a principles in which on determine to us allow will zebrafish feeding behavior is based and particularly, that what can restrict the quantity of food eaten test behavioural a established have We of development,functionandregenerationthegustatorysystem. humans and patients, considerable research healthy work in has recently behaviour been focused nutritional on in the mechanisms taste of role important during the acquired of consequence often a As substances, childhood. sweet for appetite exacerbated to linked be obesity may hand, human other in the On intake. food of patterns altered and appetite reduced through loss the permits taste chemicals. Taste sweet impairment (dysgeusia) occurs in several diseases and it instance, can give rise to weight For substances. deleterious identification reject of energy rich and nutrients, and food bitter warns against good the intake of select potentially poisonous to behavior,allowing feeding appropriate an maintaining and acquiring helps taste of sense The U784 Développement Biologie EcoleNormaleSupérieure, Paris France du Moléculaire Génétique de Laboratoire B. Boyer andF. Rosa Taste preferencesinZebrafish juveniles Behaviour 224 NEM Dpreet de Département INSERM, 295

Posters Posters 296 behavioral, pharmacological,andgeneticlevels. fishand comparable across databetween phylogeny and ontogeny and, thus, sleep opens new ways tocompare of dissecting sleep at the away mammals - including humans. In contrast to the standard in the field, this method renders sleep offers zebrafish the to method this Applying method of characterizing sleep states; a method that is based on the stability of behavioral states. The values, zebrafish andmammals. values the third, in and months; values 3 first the adult the reach and at development over found elongate are cycles rhythms sleep-wake second, circadian ages; strong all First, mammals: of that to comparable is development to used system recording and structure ontogeny.bout across sleep-wake development the that indicate behavioral data Preliminary cycle sleep-wake automated their describe time, first an the for and, zebrafish describe in cycles sleep-wake measure we experiment current the In information abouteventswithinagivenstate. transitions the about information detailed need not does one durations; species and age. constant Importantly, time the only sleep-related information needed the to calculate contrast, In development. during behavior law overall α is constant across the multiple adult mammalian in species, but switches from change exponential to power-little very of spite in duration of wake bouts; occurs this week; postnatal second the after only emerging bouts wake of behavior power-law clear a with birth, after immediately distribution are body size and metabolic rate. In neonates (rats) both sleep and exponential distribution wake where bouts an exhibit follow exponential durations bout sleep contrast, In mice). and rats, cats, (humans, species mammalian bouts exhibit a scale-free power law behavior with an exponent, wake The bout. wake individual an is exhibit t bouts where t-α wake ~ P(t) whereas, rule the bout, with power-lawdistribution sleep a individual an is t where exp(-t/τ) ~ P(t) rule the with and wakefulness. In adult humans the sleep duration of bouts of of sleep structure bouts the in exhibits revealed an been have exponential regularities distribution statistical Recently,surprising Iceland Department of Biomedical Engineering, School of Science and Engineering, Reykjavik University, K. Karlsson,I.Þ.Jóhannesdóttir, H. Arnardóttir, H.Þorsteinsson The ontogeny ofsleep-wake cycles inzebrafish 225 α and τ, thus, represent a novel way of thinking about sleep, and offer a novel, simpler τ, on the other hand, increases with age. Thus, the power-law exponent τ represents a characteristic time scale whose main determinants and α can be used to meaningfully compare sleep in sleep compare meaningfully to used be can τ α, that remains constant across α and sleep states or states sleep between τ is sleep and wake varies across varies τ Behaviour transplantation experimentsbetweenplxnD1mutantandwildtypeembryos. analysis of the PlxnD1 receptor using Se vessel patterning as an assay and the results of our cell the Similarly, elucidated. be an perform to efforts ongoing our describe we Here, uncovered. been to remain signaling PlxnD1 endothelial cells in that require PlxnD1 activity and participate the cell autonomy of this that requirement have not molecules and mechanisms biochemical precise However, the axis. dorso-ventral the along positions specific sprout from along the aorta, determines their pathfinding trajectory and limits their branching to fundamental aspects of Se vessel angiogenesis: it determines when Se vessels sprout, where they the stereotypic anatomy of the trunk’s segmental (Se) vessels. Specifically, PlxnD1 regulates four Paracrine Sema a cues from the somites on signal to the relies endothelial-specific PlxnD1 receptor to system, shape (PlxnD1). (Sema3)-PlexinD1 Semaphorin3 is pair ligand-receptor such One pattern. anatomical nervous vertebrate the like variety of attractant and repellant ligand-receptor system, interactions to achieve its highly reproducible vascular vertebrate York developing The New Medicine, Biomolecular of Institute Skirball University SchoolofMedicine, USA Program, Genetics Developmental T. ZygmuntandJ.Torres-Vázquez Molecular andcellularmechanisms ofvascular patterningby PlexinD1signaling Cardiovascular system 226 in vivoin structure-function 297

Posters Posters identify TUFsinthecardiovascularsystem. of the majority in different extent and penetrance. This pilot approach thus shows the potential of the zebrafish to defects cardiovascular specific morphology,a heart to of cells altering blood as of such accumulation them or defects vascular highly caused injections Morpholino- system. cardiovascular oligonucleotide the in expressed be to them of most found and zebrafish the of TUFs 11 tested We approach. knockdown morpholino-based a performed and genome (TUFs) for an intact activity of the heart. Therefore we searched for homologues in the zebrafish function unknown with transcripts of role functional the for screen fast a Wein interested were genes 2007). al., up-regulated et (Doss cells yielded stem embryonic murine from analysis derived were which transcriptome cardiomyocytes, in global A therapies. future develop to step first the is cardiomyocytes functional to leads that network genetic the Understanding diseases. Heart failure due to the loss of functional cardiomyocytes is one of the most frequent cardiovascular 298 1 A. Sachinidis unknown with function incardiomyocytes derived transcripts frommurineembryonic stemcells upregulated specifically to homologues zebrafish of Investigation Germany; Institute of Neurophysiology, and Center of Molecular Medicine, University of Cologne (CMMC), 227 2 Institute forGenetics, University ofCologne, Germany 1 , M.Gajewski 2 , R.Niemann 1,2 , X.Doss 1 , J.Winkler 1 , J.Hescheler Cardiovascular system 1

genome-wide analyses provide a first glimpse at the potential target genes that mediate the that genes target the potential at responses ofcardiomyocytestoNodal-BMP signalingduringcardiogenesis. glimpse first a provide analyses genome-wide signaling-deficient, or BMP overexpressing embryos using the Agilent 44K zebrafish array. These performed micro-array experiments comparing myocardial expression of wild-type, Nodal/BMP have we signaling, Nodal-Bmp by behaviors cardiomyocyte of control the Toelucidate further tissue controlling in essential are signaling polarization andmorphogenesis. Nodal and In altogether.BMP that formation causes show tube BMPexpression we heart studies, or of functional Nodal inhibition or L/R asymmetric involution medial of tissue myocardial more randomized Disruption within field. maturation heart epithelial the andof positions positions lateral affects in signaling Nodal-BMP migration of cardiomyocyte positions Modulation medial shapes. in epithelial display cardiomycytes cuboidal-to-columnar motile, field acquire are heart and the shapes of cell positions squamous lateral mesenchymal-like more within cardiomyocytes Whereas tube formation. cardiac of stages initial the during tissue occur rearrangements cellular dynamic epithelial that show We and migration cell in modulatingsuchcellularresponses. including behaviors cell morphogenesis. regulates Presently, the challenge is to signaling elucidate the exact roles of Nodal each signaling Bmp pathway that suggests evidence and emerging newly processes, specification fate cell in implicated mainly been have factors these While cardiogenesis. in involved events morphogenetic tissue and cascade signaling Nodal-BMP a via signaling L/R between link the elucidate to begun has zebrafish in work asymmetrically. Recent develop to organ first the is heart the vertebrates, In Max DelbrückCenter (MDC)forMolecularMedicineBerlin, Germany S. Abdelilah-Seyfried, J.Veerkamp, andF. Priller Heart Tube Zebrafish Formation during Morphogenesis and Polarization Tissue controls Signaling Left/Right Cardiovascular system 228 299

Posters Posters 300 both in arteries cranial these of development the to contribute that sprouts angiogenic vein-derived lethal forming veins, neighboring arteriovenous shunts. Interestingly,to the connect conduits that by aberrantly form these arteriovenous that shunts are arteries actually cranial caused enlarged vessels: disorder cranial of set particular a of vascular level the at phenotype vascular HHT the of hallmarks the dominant displays autosomal zebrafish type lethal The AVMs.and telangiectases develop Telangiectasia HHT2 potentially Hemorrhagic rare, Hereditary receptor, I type TGF-ß endothelial-specific for a the of loss heterozygous model is zebrafish HHT2 malformations a arteriovenous 2. using of development are to leadwe that (AVMs), pathways the into insight gain To ofBiological Sciences,Department University ofPittsburgh, USA P. CortiandB.L.Roman Vascular remodelingisregulatedby alkiinresponsetobloodflow in stabilizingvesselcaliberattheonsetofflow. these results suggest that in manifestation of the only not role a critical plays flow flow. Blood blood increased to response determined, adaptive an genetically are but not are shunts arteriovenous as well as number that cell suggests in increases heartbeat later lack that embryos from evidence and development the in later occurs increase further A vessels. specific these within number cell and of diameter in increase an as lesion vascular the through flow the blood with concomitantly initiates that demonstrates time over malformation this of development of analysis in but forms, 229 and wt vbg mutants. Normally these connections are severed once the arterial system arterial the once severed are connections these Normally mutants. vbg embryos, at least one of these transient connections fails to regress. Detailed regress. to fails connections transient these of one least at embryos, vbg phenotype, but also in regulation of alk1 expression is regulated by blood flow and that Alk1 might function -positive cranial vessels, and manifests and vessels, cranial alk1-positive ( beauregarde violet mutant, alk1 alk1 expression. Taken together, Cardiovascular system . Patients affected by affected Patients ALK1. vbg), vascularization inZebrafish. toh the highest doses of kdr. Furthermore, tail blisters were observed in a significant percentage of embryos injected with by or embryos transgenic tg(kdr:EGFP) vessels (ISVs) intersomitic appeared thinner in diameter even though Also, no delay in ISVs development edema. was observed in pericardial and circulation blood of lack in results is injection expression of pattern vascular of Down-regulation This hpf. vein). 30 until cardinal maintained posterior and hpf, aorta (dorsal 26 trunk At the cells. of precursor endothelial the representing likely z-edg1 expression is maintained in the CNS and becomes apparent also in the axial vasculature embryo, the of part posterior RT-PCR analysis revealed that revealed RT-PCRanalysis various whole-mount in and RT-PCRanalysis by development of angiogenesis stages different at and maturation vascular the cloned we zebrafish, in development during invasion, and of role survival, the assess cancer.to including order conditions, In pathological and physiological growth, cell of and Sciences phosphate (S1P), a blood-borne Biomedical lysophospholipid with important sphingosine-1- implications in for the receptor modulation cell endothelial an of is (edg1) gene-1 differentiation endothelial The Department Immunology, and Pathology General Biotechnology, University ofBrescia, Italy of Unit C. Tobia, S.Nicoli,Buraschi,G.DeSena,andM.Presta Characterization oftheendothelialdifferentiationgene-1(EDG1)inzebrafish Cardiovascular system stage, a stripe of 20-somite stripe the a At somitogenesis. stage, during (CNS) system nervous central the in present widely is observations, previous with agreement In (hpf). post-fertilization hours 15 from and ko157 . At present, experiments are in progress to assess the role of z-edg1 z-edg1 morpholino, as already described for the S1P signalling mutants mil, + cells is apparent in the midline and extends through the trunk and trunk the through extends and midline the in apparent is cells is expressed throughout zebrafish development zebrafish throughout expressed is z-edg1 hybridization for the vascular endothelial marker endothelial vascular the for hybridization situ in expression following expression z-edg1 ortholog and we studied its expression its studied we and ortholog z-edg1 230 hybridization. situ in z-edg1 morpholino z-edg1 mRNA z-edg1 during tumor starting edg1 301

Posters Posters 1 302 D. Epting The Rac1regulatorELMO1controlsvascular morphogenesisinzebrafish Cancer Research CenterGerman the (DKFZ–ZMBHand Heidelberg Alliance) of Heidelberg, University Mannheim,Mannheim,Faculty Medical Germany,the of Biology Vascular studies using specific morpholinos restricted vascular and neuronal expression vascular during early the zebrafishin development.role Loss-offits function for studies hybridiziation situ In but elusive. remains system systems, biological different in migration cell during activation its The ELMO1/DOCK180 complex forms a guanine nucleotide exchange factor for Rac1, regulating of Rac1 of angiogenic of intracellular proteins modulate the activation of Rac1; second, what regulation are the upstream regulators which about known is the little First, unanswered. remain vasculature adult during and embryonic the in Rac1 functions of function and regulation essential the regarding questions performs important several Todate, processes. Rac1 GTPase small The of Developmental Biology, InstituteBiology1, University ofFreiburg, Germany cascade regulatingtheformationofvasculatureinzebrafish. signalling novel a identified have data our Thus cells. endothelial in activation Rac1 to leading that showing by substantiated and complex, ELMO1/DOCK180 the of interaction functional the regulate Unc5B ELMO1/DOCK180 and Netrin-1 biochemically were the data functional These of ELMO1. for activators seen as upstream as Unc5B receptor complex. Loss-of-function experiments for Netrin-1 and Unc5B display overlapping its phenotypes and Netrin-1 identify could Moreover,we duct. thoracic the of development the and vessel parachordal the vessel, formation of the vasculature, including intersomitic vessels, the dorsal longitudinal anastomotic Center for Biomedicine and Medical Technology Mannheim (CBTM), Joint Research Division Research Joint (CBTM), TechnologyMannheim Medical and Biomedicine for Center 231 , and third, what are the functions of these regulators in the vasculature the in regulators these of functions the are what third, and vivo, in 1 , B.Wendik 2 , K.Kern 1 , W. Driever against elmo1 severely impaired in 2 , J.Kroll 1 revealed a temporal and spatial and temporal a revealed elmo1 Cardiovascular system tg(fli1:EGFP) embryos the 2 Department Department in vivo? in lead tothisphenotype. that events molecular the understand and tissue this in protein zSARA1 the of distribution expression and the analyse to interesting be would It observe. we that ventricle the into valve A/V missexpression might also be the cause for the expansion of the cuboidal endocardium from the BMP4 While formed. properly not expression is still is present in the valve A/V valve, Notch1b (A/V) is missexpressed in atrioventricular this tissue. This in the Notch1b that zSARA1, us of showed absence analysis This the valves. cardiac the as well as chambers cardiac the and cells the analysed have We circulation. blood no had morphants the that us showed morpholinos with function zSARA1 of down knock The cell asymmetric during signaling directional Notch division intheSOP (Coumailleau,Nature2009). labels it that and 2006) Science (Boekel, precise transmission of TGFbeta-signaling levels during symmetric cell division in the wing disc Cell 1998). Furthermore, studies in our lab have shown that drosophila SARA is required to allow as a TGFbeta signaling adaptor that brings Smad2 to the activated TGF-beta receptor (Tsukazaki, It has been show in cell culture that SARA is an endosomal protein with a FYVE domain, and affect theatrioventricularvalvedevelopment. acts a study on the TGFbeta signaling adaptor, biological zSARA1, we serendipitously saw that this protein can are chamber cardiac defects are still the the most common human congenital abnormalities. In our of lab, while performing septation and processes tightly regulated by valves several signaling pathways. However, valve and cardiac chamber cardiac of morphogenesis The University ofGeneva, Switzerland C. CamposandM.González-Gaitán Regulation ofcardiacdevelopment by zSARA1 Cardiovascular system morphology of blood vessels, the presence of blood of presence the vessels, blood of morphology 232 303

Posters Posters M. Arese 1 ihrh, ert-ia, Italy; Segrate-Milan, Richerche, 304 Rissone A. Synergistic roleofNeurexinsandNeuroliginsinvascular embryonic development Angiogenesis, Institute for Cancer Research and Treatment, University of Torino, Italy; of Biology, University of Milano, Italy; synergistic rolebetweenbeta-NeurexinsandNeuroliginsduringangiogenesisinvivo.· :DsRed) Tg(gata-1 recapitulates the observed circulatory and vascular abnormalities suggesting a and Tg(fli-1:eGFP) in Neuroligin and beta-Neurexin for MOs translation-blocking produced the of low doses Neuroligin of coinjection and The effects. morphological beta-Neurexin gross any for without phenotypes MOs vascular specific defects, vascular obvious without larvae, and embryos of activity locomotor impaired knock-down alpha-Neurexin the While embryos. :DsRed ) Tg(gata-1 used we morphants, the of integrity moreover,system vascular the analyze to Tg(flk1:GFP) and Tg(fli-1:eGFP) by RT-PCRwith in injected weredetermined been have MOs Single MOs sites. target MO the flanking exons the on ofsplice-blocking designed primers efficiency and specificity the and phenotypes vascular similar very produced MOs of targetedcouple each Each of gene. splicing the and translation the block respectively in to designed defects been have vascular MOs zebrafish. different specific Two induce alpha-Neurexins not but beta-Neurexin and Neuroligin of alternative of sites isolated we the seven differentzebrafish Neuroliginsrecently and here we showMore that functional knock-down contains oftranscripts. specific forms different and of thousands Like form to rise give (beta) can counterparts. that short (AS) splicing a human and (alpha) the long a to the of drive that synthesis promoters respect alternative two presents in gene of Neurexin zebrafish structure steps and analyzedeach genes adhesivehuman and genes early relationships Neurexins the zebrafish phylogenetic in six thantheir the rather isolated we transmission Previously, synaptic formation. ofsynapse modulation alpha- the three in the of role expression the of knockdown Neurexins or Neuroligins 1-3 transmembrane Thein the mouse demonstrates that these proteins have synaptic a synaptogenesis. fundamental of in families involved two proteins constitute Neuroligins and Neurexins mammals, In involved inembryonicvasculardevelopmentandbloodvesselremodelingzebrafish. have explored this hypothesis and found that specific isoforms of Neurexins and Neuroligins are receptors. the of Eph none known systems, key synaptic proteins these has been by theirso far shared shown to participate molecules in blood of vessels functions. and number We growing still and Ephrins high the Despite and Slits-Robos Semaphorins-Plexins-Neuropilins, Unc5s, signaling specific been functions.by orchestratedmolecules in their is the have extracellular cells environment, which regulating endothelial belong systems to and four cues distinct axons classes: Netrins-DCCs- nervous of molecular guidance common the and example of For vascular discovery the the of between light relationships in re-evaluated the decade last the In University ofMilano, Italy eatet f nooia Sine, nvriy f oio Iay Dvso o Molecular of Division Italy; Torino, of University Sciences, Oncological of Department 233 1* 1§ , E. Foglia E. , 2 , L. Sangiorgio L. , fish lines to visualize the development of the vascular system; vascular the of development the visualize to lines fish 4 eatet f imlclr cecs n Biotechnologies, and Sciences Biomolecular of Department 3 3 , S. Cimbro S. , Istituto Tecnologie Biomediche-Consiglio Nazionale delle 2 , M. Beltrame M. , 4 , F., Bussolino Cardiovascular system 1* , F., Cotelli 2 Department Department 2* and 1 F.C. Simões Fgf favours acardiacfateattheexpenseofhaemangioblastprogramme inzebrafish Cardiovascular system University, John Radcliffe Hospital, Headington, UK;Headington,Hospital,University, Radcliffe John in theadultheart. cells stem of potentiality the for and cells pluripotential from cardiomyocytes of derivation the for implications have observations These programmes. and endothelial and programme blood the of cardiac repression the of elaboration full the for necessary are activities combined their fgf3 andfgf8,separatelytogether, wedemonstratethattheyarethekeyFgfligandsand of expression down knocking By programme. haemangioblast the represses concomitantly and programme heart the application induces Fgf that temporal show we By signalling, Fgf of field. inhibitor molecule heart small a of the into haemangioblasts anterior the of recruitment to the leading evolution during extended been have could influence of domain whose characteristics signal a the of has (Fgf) factor growth fibroblast that show further We fates. cell two these also inhibits haemangioblast development, indicating a cross-antagonistic relationship between is programme haemangioblast the that inhibitory indicated to the development of the cardiac programme. have Here we show that a cardiac regulator experiments function of loss possibly and embryos, gain mammalian in found not haemangioblasts representing an evolutionary antecedent anterior of the second heart of field. Consistent with such a population notion, a have to be present in zebrafish, possibly explaining its smaller two-chambered heart. Instead zebrafish appears which of one only fields, heart embryonic two from fashioned is heart mammalian The and Biomedicine, Center andCell forNeuroscience Biology, University ofCoimbra, Portugal R Mlclr amtlg Ui, eteal nttt o Mlclr eiie Oxford Medicine, Molecular of Institute Weatherall Unit, Haematology Molecular MRC 1,2* , T. Peterkin 1,* , R.Patient 1 2 PhD Programme in Experimental Biology Experimental in Programme PhD 234 305

Posters Posters 306 acting be may upstream ofVEGF. sulf1 brain ThisworkhasbeenfundedbytheBritishHeartFoundation. the in if and activity VEGF on influence sulf1 reflects programming be due to a reduction in VEGF expression. We are now investigating whether the altered arterial reduced circulation and defective formation of the central arteries in the brain that appeared to the Flt4 venous marker, suggesting that sulf1c has a role in arterial programming. There fish, was of expression in also increase an with areas, these in reduced/lost severely is 26hpf at EphrinB2 and Delta Tg[fli1EGFP-gataDSred] Notch, markers arterial of Expression aorta. distal the in of occlusion to leading formation, knockdown morpholino confocal imaging Following of the Sulf1c morphants at 26hpf onwards revealed defects angiogenesis. in the caudal artery points around time at expressed temporally be to transcripts sulfs zebrafish three all show results Our on thevascularroleofHS6-O-sulphationfine-tuningbysulfenzymes. focus recent our to led This branching. vascular directed VEGF reduced HS, 6-O-sulphates that receptors. In zebrafish embryos we found that knockdown of the 6-O-sulphotransferase 2 gene, shown that HS is important for both gradient formation of VEGF and its interaction the key angiogenic growth factor,with vascular endothelial growth factor (VEGF), to HS. signalling It has been previous biochemical studies have demonstrated that 6-O-sulphation is critical for the binding of the HS sugar chains affects in their binding to and protein ligands, including surface angiogenic growth factors. cell Our from pre-existing the vasculature, (angiogenesis). Variation on in the sulphation patterns of (PGs) the complex proteoglycans (HS) extracellular environment sulphate is critical to heparan many processes including of the growth of presence new blood The vessels Cardiovascular Group, University ofManchester, Core Technologies Facility, Manchester, UK S. E.Stringer,B.Gorsi,X.Ma,T. Chico,F. Liu,andR.Patient functioning ofthebrain vasculature for and programming arterial for critical is sulfi1 by structure sulphate heparin of tuning Fine 235 Cardiovascular system CT-2005-018630 Repair‘Heart ‘ andtheDFG-fundedgraduatecollege1048 „Organogenesis“. COB2006,the from EMBO bygrants ASTF96-2007,LSHM- funded was project work FP6 This in genes these cardiac conductiontissuedevelopmentbecameapparent. of requirements differential also however functions, overlapping have Popdc2 but showed an overall reduced beating rate indicative of a sinus bradycardia. Thus, an AV block in of presence the revealed microscopy illumination plane selective and line transgenic indicator sinus rhythm and atrial contractility appeared normal. Recordings of calcium transients while using In an rhythm atrio/ventricular 3:1 development. and 2:1 with contractions muscle ventricular irregular observed tail we aberrant and Both, edema embryos. pericardial developed zebrafish in approach morpholino-based cardiac pacemaking is evolutionary conserved. We addressed this possibility using an antisense os hat I ti suy w teeoe se wehr h rqieet of requirement the whether asked therefore we study, this In heart. mouse nvriy f nsrc, Austria, Innsbruck, of University T. Brand . . Kirchmaier C. B. Function ofthePopdc genefamilyincardiacconductionsystemdevelopment Cardiovascular system skeletal muscle. In mice l The Singapore. 1 f aiona Sn rnic, USA, Francisco, San California, of el n Dvlpetl ilg, nvriy f urbr, emn, Germany, Wuerzburg, of University Biology, Developmental and Cell ) gene family is predominantly expressed in heart and heart in expressed predominantly is family gene Popdc( ) containing domain Popeye 1 Popdc2 morphants. Surprisingly, 1 . Günthner F. , oss of Popdc1 and 1 T Schwerte T. , 3 4 eatet f iceity n Bohsc, University Biophysics, and Biochemistry of Department eatet f ilg, ainl nvriy f Singapore, of University National Biology, of Department Popdc2 Popdc1 morphants did not develop an AV block 2 J Huisken J. , affected sinus node function in the postnatal and Popdc1 3 C Winkler C. , 236 2 nttt o Zoology, of Institute morphants Popdc2 morphants Popdc2 4 DYR Stainier D.Y.R. , Popdc Popdc1 and genes 307 for 3 ,

Posters Posters 308 2 D. Bournele Zebrafish modelsofcardiacdevelopment anddisease 1 the meeting. during presented be will gene this of function and mapping phenotype, the on data latest The on MGH). developmental the (48cM 15 group into linkage on maps mutation The functions formation. valve cardiac control it that pathways how elucidating and phenotype the for responsible gene mutated the cloning and identifying on working currently are tract. We outflow the at residing cells elastin-positive less embryos, type wild to compared we have, addition, mutants our In that cells. shown endocardial have cuboidal of layer single a by lined is canal atrioventricular the that showed we and mutants these in development valve the resolution cellular at analyze to microscopy confocal use we parallel, In stenosis. tract outflow an to due edema pericardial exhibit but function, myocardial normal have and hpf 72 at type wild the morphologically appear and embryos mutant tract The fertilization. outflow post hours functional 55 at a finally stops initiated, develop normally circulation, to fail embryos mutant Homozygous humans. Inoneof in of valvedevelopment. result phenotype stenosis aortic an to a similar (hpf), cardiac fertilization post hours as 72 at stenosis tract outflow canal atrioventricular the ofdefective at regurgitation blood blood observed anindication retrograde we (s411), lines a as these exhibit lines heart mutant the These labs). through Baier flow and (Stainier UCSF the in screen genetic large-scale forward a To during identified mutants the of advantage take we effect, that is tofindouthowheartvalvesformandfunctionthroughoutthelifeofvertebrates. forward genetics and study of gene function at the and cellular level. The atrial long-term aim of our efforts etiology the of possibility the offers Zebrafish defects. canal atrioventricular to of and defects septal ventricular linked been has development cushion endocardial mammals, In for the regulation of the initial stages of zebrafish atrioventricular endocardial cushion and formation. interactions endocardial-myocardial that it relies involves on heart function. canal We also atrioventricular showed that Notch the and calcineurin signaling of are required differentiation microscopy. confocal the and that immunofluorescence showed lines, We transgenic gfp using the cellular architecture of the zebrafish heart during stages ofpathways atrioventricularconverge valveto development, orchestrate valve development. We have prevent published retrograde to a blood vertebrates of detailed flow.life the description throughout function Theseof and cells cellsendocardial from undergoderive valves elaborateCardiac morphogenesis and several signaling eeomna booy boeia rsac fudto, cdm o Ahn, Greece; Athens, of Academy foundation, research andbiophysics,Biochemistry University ofCalifornia, SanFrancisco, biomedical USA biology, Developmental 237 1 , D.Stainier 2 and D.Beis 1 Cardiovascular system Regulation of Sphingosine-1-phosphate (S1P) signaling during zebrafish cardiovascular zebrafish 1 during signaling C. Detzer (S1P) Sphingosine-1-phosphate development of Regulation Cardiovascular system ede CC, ike SA (06 Shnoie1popae niis el irto and migration cell inhibits Developmental development. Sphingosine-1-phosphate Biol. cardiac 291, 264-277 during (2006) transformation cell mesenchymal S.A. to endothelial Rivkees C.C., Wendler and non-lymphoidlesions. PLoS ONE4(1):e4112 lethality early prevents yet function system immune modulates lyase 1-phosphate sphingosine VogelP, Donoviel MS, ReadR, GM,Hansen HazlewoodJ, al.et of inhibition Incomplete (2009) was which heart, the in phenotype reproducible anddosedependent. a to led knock-down Morpholino spectrometry. mass by S1P and sphingosine ceramide, of concentrations lipid determine we addition In knock-down. by pattern expression its analyze out mice (Vogel et al., 2009). To investigate the role of SPL in zebrafish heart morphogenesis we interstitial cellularity and vacuolation in the myocardium. S1P level, were elevated in SPL knock- 2006). In addition SPL knock-out mice displayed histopathologic lesions in (WendlerRivkees, morphogenesis and heart the for important is S1P of heart levels local Tightof control by increased and cellproliferation.TheS1P lyasedegradesS1P tophosphoethanolamineandhexadecanal. ceramide- and sphingosine lead to growth arrest and cell death essential whereas are S1P SPLbecause important, is very is which cell, the S1Pinside the rheostat and of maintenance needed for regulators for SPPs S1P.growth SpKs, of precursor a also is that ceramide form to be N-acylated then can which sphingosine precursor the to back S1P dephosphorylate phosphatases S1P The (SPL). lyase S1P the by and (SPPs) phosphatases S1P the by sphingosine degraded is S1P by (SpK). kinases catalyzed sphingosine, of phosphorylation the by produced is ligand S1P The differently are receptors These expressed and Genes“. coupled to Differentiation various „Endothelial G-proteins, so as numerous downstream known signals can also be are regulated. which an plays and sphingosine development and immunity. from heart S1P maturation, is a vascular ligand of derived fiveangiogenesis, G-protein coupledincluding S1P receptorsprocesses (S1P1-S1P5) mediator developmental in role lipid important a is (S1P) Sphingosine-1-phosphate Pharmacology/ZAFES, Johann Wolfgang GoetheUniversity, Frankfurt amMain, Germany Max-Planck-Institute for Heart- and Lung Research, Bad Nauheim, Germany; 1 , K.Scholich 2 andB.Jungblut hybridization and its function by morpholino mediated morpholino by function its and hybridization in-situ 1

238 2 Institute for Clinical 309

Posters Posters 310 S. Hundt,D. Y.R. Stainier The roleofatpbindingdomainprotein4inheartdevelopment ofzebrafish a-lnkIsiue o Hat ad ug eerh Bd ahi, Germany; Nauheim, Bad Research, Lung and Heart- for Max-Planck-Institute development. Development 132: 4193-4204. (2005) al et Beis partners ofourproteininteresttoidentifythemolecularprocessesthisisinvolvedin. the sub-cellular distribution of the protein. Furthermore we are searching for possible interaction the molecular function of ATPDB4. Therefore we are investigating the gene expression as well as mutant. This phenocopy can be rescued by injection of the ATPBD4-mRNA. We are interested in human the of the to similar lesion molecular the highly identified not is ATP.have bind we might far which it So that suggests protein, orthologue, this of sequence The function. unknown with protein (ATPBD4),novel 4 a ATPprotein the domain encodes binding gene This gene. single a containing group17 linkage on region a to mutation the maps mutant the of cloning Positional well asattheatrio-ventricularjunction. as ventricle and atrium the in cells myocardial of configuration and number,shape, abnormal and sinus venosus, and edema in the heart region. On a cellular level, mutant embryos show an atrium the between regurgitation blood ventricle, collapsed a to leads that identified was s272 In a genetic screen for heart morphogenesis mutants (Beis better in help could understanding congenitalheartdiseases. development heart during processes cellular these of control genetic the Understanding heart. the of function normal the for essential are configuration cell and shape known that specific patterns of proliferation and differentiation as well as specific changes in cell of ballooning, looping, and the formation of the atrio-ventricular and the sinoatrial junctions. It is result a is heart mature The undergoeschanges. heart many the development in early very tube The embryonic development of the heart is a highly complex process. Starting from a linear heart andBiophysics,Biochemistry University ofCalifornia, SanFrancisco, USA 239 mutation in this gene but its morpholino knock-down leads to a phenocopy of the of phenocopy a to leads knock-down morpholino its but gene this in mutation s272 Genetic and cellular analyses of zebrafish atrioventricular cushion and valve and cushion atrioventricular zebrafish of analyses cellular and Genetic * , andB.Jungblut et al, 2005) a recessive lethal mutation Cardiovascular system * eatet of Department of blood and endothelial development. Studies in mouse chimaeras have shown that Flk1 lack complete a Targetedto vasculature. due lethality embryonic early to leads Flk1 of deletion formation of blood and endothelial cells in the extra-embryonic yolk sac and the intra-embryonic factor (Vegf), Flk1, is found in cells that leave the posterior primitive streak and contribute to the In the gastrulating mouse embryo, expression of the receptor for the vascular endothelial growth Institute ofGenetics, SchoolofBiology, University ofNottingham, Queen’s MedicalCentre, UK P. Lau,K.McMahon,M.Gering A cell-autonomousrequirementfor Vegf signallinginangioblastdevelopment Cardiovascular system downstream ofthehaemangioblaststageduringvertebrateembryogenesis. that Vegf signalling has a suggest cell findings autonomous our role pathway. Thus, in signalling endothelial the Vegf cell of specification morphants and and development mutants zebrafish in for Vegf in endothelial cell specification/differentiation in zebrafish embryos that is not apparent trunk vasculature when our in Surprisingly, the competition with processes. wild-type whether cells. above the to Our contribute to Vegfresults fail a test signal the receive demonstrate to unable are an which cells in early that to demonstrate results role autonomous transplantations cell cell is stage signalling Vegf blastula for performrequirement to opportunity the took We of specification haematopoietic stemcells(HSCs). c) and DA, the from vessels intersomitic of sprouting b) (DA), aorta dorsal the of specification arterial a) for; required is but differentiation, cell endothelial for needed isconfined not embryo is zebrafish and the in signalling Vegf restricted that shown ismore have studies for Previous cells. progenitor expression endothelial to common Flk1 the zebrafish, haemangioblast,In cells. the endothelial of and level blood the at possibly cells, those in Vegffor role autonomous cell a suggesting cells, endothelial and blood to contribute to unable are 240 -/- cells 311

Posters Posters TD formation in the tg(fli1:EGFP) the in formation TD lymphatic development. in Sox18 of role conserved evolutionary and pivotal the confirm results Our differentiation. cell endothelial lymphatic during PCV the in levels expression different their to linked probably is to 5 dpf. Our data indicate that the single knockdown of knockdown single the that indicate data Our dpf. 5 to 3 from (PCV) vein cardinal posterior the and (DA) aorta dorsal the between zebrafish in forms in zebrafish lymphatic development. The thoracic duct (TD) is the first perfused lymph vessel that zebrafish that shown have We a lymphatichallmarkgene,andSox18directlyactivatesProx1transcription. 312 venous the Given arterial. origin particular,of than lymphatic cells, this affected In prompted us to identity.investigate more whether appears arterio-venous differentiation correct cell endothelial a venous of acquisition the with interferes knockdown double partial their development: vascular in roles redundant than play Sox18 and earlier Sox7 vein. axial away fade they and precursors, cell endothelial in mRNAs B6 pure a in lethality fetal and edema subcutaneous background. Moreover, great mouse display to Recently,shown genes. were target mice to null binding from proteins SOXF wild-type preventing by way negative dominant a in act and domain transactivation functional a lack proteins SOX18 type lymphatic in and mutations spontaneous to vessel blood hair, development. The murine in conterpart of the disease is represented by the ragged phenotype, due defects factors. combining transcription (HLT), HMG-box Lymphedema-Telangiectasia Sry-related of SoxF-group the to Mutations in belong -18 and -17 Sox7, diScienzeDipartimento BiomolecolarieBiotecnologie. Università degliStudidiMilano, Italy S. Cermenati,Moleri,R. Amodeo andM.Beltrame Sox18 regulateslymphaticdevelopment inzebrafish eeoig lo vses Noteworthy, vessels. blood developing 241 SOX18 have been associated with dominant and recessive forms of Hypotrichosis- and presenting signs of lymphatic dysfunction. The ragged- The dysfunction. lymphatic of signs presenting and Sox18 Sox18 is expressed in a subset of cardinal-vein cells before and sox7 y1 line. The different impact of the two genes in TD formation TD in genes two the of impact different The line. rncit ae eetbe ale than earlier detectable are transcripts sox7 r c-xrse i te nohla cls of cells endothelial the in co-expressed are sox18 , but not of not sox18but , sox18 and Cardiovascular system mRNAs in the in mRNAs sox18 , greatly impairs sox7greatly , sox7 play a role also Sox18-/- Prox1 sox18 , netd mro dsoitd 4 ad 4hus fe taslnain 2hf n 4hf stage) 48hpf hCD34 and of differentiation (28hpf endothelial transplantation showed after 44-hours and 24- dissociated embryos injected vascular phenotype caused by caused phenotype vascular Time-lapse confocal analysis performed 2 hours after transplantation showed circulating hCD34 of developingTg(fli1:EGFP)transgenicembryos(48hpf),priortoimmunesystem development. zebrafish embryo. differentiation mechanisms toward the endothelial lineage and their angiogenic properties hCD34 with Orange Cell Tracker™ dye (Invitrogen). hCD34 (Invitrogen). dye Tracker™ Cell Orange with KDR (CD3, markers CD38, lineage CD45 hematopoietic and whereas CD48) CD146), were CD105, not (KDR, affected. markers endothelial Further,of levels immunofluorescence analysis showed (hCD34 di Milano-Bicocca, Italy; U. Fascio cells in developing vessels and no evidence of vascular occlusions. Further, at 12 hours after hours 12 at Further,injection occlusions. , vascular some of hCD34 evidence no and vessels developing in cells zebrafish blastula and hCD14 and blastula zebrafish 3 Human CD34 Cardiovascular system above changeswerenotobservedinhCD14 In other experiments hCD34 1 Conclusions: the embryodonotseemtoaffectvasculardevelopment. P.Vella hCD34 Results: and Methods development aftertransplantationintothezebrafishblastulaandembryo. raised the possibility that hCD34 that possibility the raised as well as enhanced and ectopic angiogenesis at the level of the subintestinal vein; these results Objective: The objective of cells the stem present study of was study to the determine for whether system differentiation andplasticity.human model (TraverCD34 interesting etal.,2003;Traver and etal.,2004). novel a is embryo zebrafish the into (Lawson sprouting cells endothelial of in confocal imaging by observe example, for to as, vasculature possibility embryonic the the of development including the angiogenesis,real-time and vasculogenesis vertebrate of study for advantages of number a provides ) rerio (Danio embryo zebrafish The Background: this hypothesis it was found that hCD34 that found was it hypothesis this Centro Interdipartimentale Microscopia Avanzata Italy; (CIMA), Milano, -IRCCS,Università di Monzino Milano, Cardiologico Italy; Centro + and VE-cadherin and + 1 + - injected embryos exhibited , O. Pozzoli O. , ) cells differentiate toward the endothelial lineage and contribute to new blood vessels blood new to contribute and lineage endothelial the toward differentiate cells ) 3 , A. Biondi h peet td dmntae te vltoay osrain f hCD34 of conservation evolutionary the demonstrates study present The + CellDifferentiationtoward theEndothelialLineagein Zebrafish Embryo 1 4 , G. Iaffaldano G. , , M.C.Capogrossi + + human cells localized in the dorsal artery and in the cardinal vein. The vein. cardinal the in and artery dorsal the in localized cells human cells had incorporated into the vessel wall. One day after transplantation, 5 Istituto Dermopatico dell’Immacolata -IRCCS,Istituto Dermopaticodell’Immacolata Roma, Italy + and hCD14 + - cells were isolated from cord blood and subsequently labelled subsequently and blood cord from isolated were cells cells were used as controls. Flow cytometry (FACS) analysis of analysis (FACS) cytometry Flow controls. as used were cells knock-down. On the contrary,hCD14 the On Vegfcknock-down. + cells may synthesize angiogenic factors. In agreement with agreement In factors. angiogenic synthesize may cells altered blood vessels sprouting in the growing tail vasculature 1 , M. Lacovich M. , 5 , andM.Pesce + - cell injection into the zebrafish blastula rescued the rescued blastula zebrafish the into injection cell cells (500-1000) were injected into the sinus venosus - cell–injectedzebrafish. + cells, as demonstrated by increased expression increased by demonstrated as cells, ., 2002). Furthermore, cell transplantation cell Furthermore, 2002). al., et 1 , P.Devanna , + 2 cells (400-500) were injected into the into injected were (400-500) cells 1 nvrià el Sui i iao Italy; Milano, di Studi degli Università 1 , C. Lora Lamia Lora C. , 242 - cells injected into injected cells 2 , F.Cotelli , + 4 progenitor Università in vivo in in vivo in + in the cells 313 2 + ,

Posters Posters visual morphologyorfunction. affecting without eye the in angiogenesis inhibit that 2 identified and drugs 1500 over screened at the effect of the selected drugs on visual behaviour and retinal morphology. To date we have clips. fin looks adult characterisation in Tertiary angiogenesis regenerative and model mutant a abnormalities. developmental gross inducing in angiogenesis ectopic inhibit to ability their for screened then are stages these pass that Drugs without dpf angiogenesis. hyaloid on 5 response time-dependent and dose- a at confirming Validationinvolves hyaloid the of angiogenesis lead the of some of characterisation drugs uncovered. Primary and drug hits are identified as thosevalidation that induce defects in developmental screening, the on presented be will Data developmental of inhibitors for screen angiogenesis ofthehyaloidvasculatureinzebrafishlarvae. chemical a performed we neovascularisation, ocular inhibit that drugs lead identify to order prematurity.In of retinopathy and retinopathy diabetic Retinal neovascularisation is a pathological hallmark of debilitating forms of blindness including Microbiology Tumor and Cell Biolog, Karolinska Institute, Stockholm, Sweden Y. Cai 314 1 Kennedy B. Chemical InhibitorsofDevelopmental Angiogenesis intheEye UCD SBBS & UCD ConwayUCD & SBBS Institute,UCD College Dublin,University Dublin, Ireland 243 1

1 Y.Alvarez , 1 , N. Waghorne N. , 1 , O. Astudillo-Fernandez O. , Cardiovascular system 1 , L. Jensen L. , 2 , S. McLoughlin S. , 2 Institution for Institution 2 , 3 B. M.Hogan Dll4 suppresses Vegfc/Flt4 signallinginthedeveloping zebrafish arterialsystem Cardiovascular system 1 lymphatic vessels and isolated the mutant the isolated and vessels lymphatic We performed a forward genetic screen in zebrafish towith exquisiteprecision. identify mutants that fail to form embryonic embryo, the function of many reiteratively vessels acting molecular pathways is lymphatic expected to be regulated or regulators. Tomolecular common many orchestrate blood share the that development processes of of linked these intimately two distinct are vascular vessels, development systems pre-existing in from the vertebrate the lymphangiogenesis, and Hemangiogenesis Helsinki, University ofHelsinki, Finland flt4 upon observed phenotype hyper-branching the that found and of downstream acts flt4 that and hemangiogenesis for required To vessels. developing in function Flt4 limiting by potential angiogenic limits Dll4 is whether test signalling Flt4 that demonstrated have data Recent flt4 signalling deficient mutant but arterial hemangiogenesis occurred normally in these mutants. dll4 for mRNA encoding domain Ig inhibitory soluble or nml. e on ta increasing that found We animals. adooy Toacne, rsu Uiest Mdcl etr Rtedm Te Netherlands; The Rotterdam, Center, Medical University Erasmus Thoraxcenter,Cardiology, to ae tgte, hs dt dmntae ht l4 upess h aiiy f developing of ability the suppresses the embryoduringarterialandvenoushemangiogenesis lymphangiogenesis. Dll4 that this mechanism provides for the differential endothelial demonstrate response to a constant source thatof propose Vegfczebrafish. We in signalling in data Flt4 Vegfcdriven to respond to arteries intersegmental these together, Taken induction ofectopicsproutingintersegmentalarteries. Molecular Cancer Biology Laboratory, Department of Pathology, Haartman Institute, Biomedicum Hubrecht Institute-KNAW & University Medical Centre, Utrecht, The Netherlands; vegfc/flt4overexpressed we signalling, signalling mutants. Furthermore, we found that the injection of hyper-branching phenotype. To test whether loss of loss whether Totest phenotype. hyper-branching 1 , R.Herpers dll4 1,2 , M.Witte in zebrafish, we knocked down knocked we zebrafish, in n h cnet f depleted of context the in vegfc 1 , H.Duckers vegfcmRNAby in injection . Positional cloning identified cloning Positional expando. 2 , K. Alitalo ee oh aal o rsun the rescuing of capable both were Flt4 sensitises intersegmental arteries intersegmental sensitises dll4 in the in dll4 3 , S.Schulte-Merker knockdown was absent in absent was knockdown dll4 244 vegfc dll4 dll4 mutant background mutant flt4 targeting morpholinos morpholino-injected e t a synergistic a to led 2 1 as a as expando Experimental Experimental 315

Posters Posters ee epesd n h eiadu is epicardium the in expressed genes species might therefore contribute to the understanding of their pluripotency. One of the earliest other in (EPDCs) s cell derived epicardial of fate the on Data debate. under currently is which reported, been has lineage cardiomyocyte and the to contribution cells additional an whereby blood chick, obtained in results vessels, the confirm coronary partially the mouse the of in performed cells experiments Recent muscle fibroblasts. smooth and endothelial to rise giving myocardium, the invade and transition epithelial-mesenchymal undergo cells epicardial splanchnic some of protrusion chick the In it. covers that layer epithelial an a forming myocardium, the of surface the to adhere proepicardium, the cardiomyocyte and delaminate from cells Proepicardial venosus. sinus promoting the of base the at allocated origin mesoderm by derives and epicardium cells progenitor The of proliferation. source a as development heart during roles important exerts which myocardium, the enclosing layer epithelial an is epicardium The morphogenesis, CABD, Sevilla, Spain analysis oftransgeniclineswillbepresented. and generation the on progress Recent larvae. zebrafish in expression gene epicardial-specific the Conserved of line. elements transgenic epicardial-specific noncoding an generating are we this, For of cells. progeny epicardial the study to and development heart on ablation epicardial of effect the analyze to vivo, in formation epicardial analyze to tools the establishing Weare development. epicardial of processes study to model animal zebrafish the of advantages the using in interested Weare expression ofepicardialmarkergenes. A adults. in re- to leads heart resection apex ventricular that growing finding the by the supported is regeneration to during role contribute cells epicardial that show experiments homeostasis, DiI cardiac during since role a play to suggested been has epicardium the species, this In of function Recently,and expression the buds. limb and system mutants displaydefectiveepicardiumformation. nervous central gonads, pancreas, pronephros, the include 1 316 N. Mercader Generation andanalysisofanepicardial-specifictransgenic lineinzebrafish the proepicardium and switched off upon EPDC differentiation. Other sites of sites Other differentiation. EPDC upon off switched and proepicardium the adoaclr eeomn, NC Mdi, Spain; Madrid, CNIC, Development, Cardiovascular 245 1 , J.M.GonzálezRosa eoi rgo ae en tse fr hi aiiy o drive to ability their for tested being are region genomic WT1 1 , A. Ariza im tmu 1( tumour Wilms has been shown to be conserved in the zebrafish. the in conserved be to shown been has wt1 2 , J-L.Gómez-Skarmeta 2 eatet f ee euain and regulation gene of Department . t epeso i tre o in on turned is expression Its WT1). Cardiovascular system 2 , V. Martín WT1 1 , M.Torres expression null Wt1 1

zygotic mekk3bmaycontributetofunctionalintersegmentalvesselformation. mekk3b may contribute to functional dorsal aorta and posterior cardinal vein formation, and that posterior cardinal vein, but caused defects in the segmental vessels. Thus, we think that maternal In vein). contrast, cardinal posterior injection and of aorta SD-mekk3b (dorsal trunk MO the that of blocks vessels splicing axial major did in not abolished show was defects in dorsal aorta and ATG-morpholino oligonucleotides (MO) into fertilized eggs. Blood flow of the To determine whether knockout ofmouseMekk3causesdefectsinthebrain,heart,trunkandplacenta. gene is a serine-threonine kinase that belongs to the MAP3K family. It has been reported that the 1 is integrated into an intron of the of intron an into integrated is expressed is GFP in line, the SAGFF27C;UAS:GFP heart, the posterior In line. cardinal SAGFF27C vein a and identified intersegmental we vessels. screen, trap The gene gene the trap construct the developed we Previously, formation hasnotbeenfullyelucidated. vessel blood functional of mechanism molecular The vessels. sprouts intersegmental the in begins flow angiogenic posterior primary blood then and the vessels, primary the to from connections make trunk, sprouts emergesecondary The vein. cardinal sprouts angiogenic zebrafish secondary then developing aorta, dorsal the the from emerges In gases, fluids, cells. of transport for and essential are macromolecules that networks complex forms system vascular The Morioka, Japan A. Urasaki of involvement reveals formation approach trap gene The Cardiovascular system National Institute of Genetics,Yata,1111 of Institute Japan;Mishima,National Shizuoka 1 , S.Isogai 2 , E.Kimura mekk3b plays an important role in the blood vessels, we injected -mediated gene trap method in zebrafish. In the course of course the In zebrafish. in method trap gene Tol2-mediated 2 , J.Hitomi gene and traps the transcript of transcript the traps and gene mekk3b 2 andK.Kawakami n h fntoa bod vessel blood functional the in mekk3b 1 246 2 Iwate Medical University,Medical Iwate . The mekk3b. mekk3b morphant mekk3b mekk3 317

Posters Posters 318 A. E.Reyes Funtional interaction betweenHdac9andHif-1alfainheartdevelopment ofzebrafish 1 nrs el, atao Chile; Santiago, Bello, Andrés Supported bySupported FONDECYT1095128to A.E.R. the DNA,activatingcardiacgenes. Hif-1alfa to the DNA, for of the contrary the absence binding of Hdac9 facilitate the the binding of Hif-1alfa to avoiding compactization chromatin induces Hdac9 that propose We Hif-1alfa. knockdown of Hif-1alfa. These results suggest that Hdac9 may have a functional interaction with double knockdown by induces a evaluated down-regulation was of cardiac effect gene, The like results proteins. obtained both by the to single knockdown To answer if Hdac9 regulated the action of Hif-1alfa to activated cardiac genes we did a double Hdac9. of knockdown the of effect opposite the expression, gene cardiac of regulation down a generated factor this of function of lost the that low is this under for evidence active supporting tension, is oxygen that factor transcriptional a (Hif-1alfa), hypoxia-1alpha by induced factor transcriptional a allows Hdac9 of knockdown the factor to bind to that the DNA. One potential propose candidate to be Weregulated by Hdac9 is hypoxia. the transcription to exposed are hand2, and as expression of gene regulation down differentiation by malformation cardiac generate Hdac9 of function of fail the that show the mRNA in the heart, this result were confirmed by RT-PCR.of expression detected have we Previousand hpf, 72 resultthrough hpf 0 fromfrom express is ourhdac9 that found we laboratory our focus we Here DNA. interesting in the action the of Hdac9 in the cardiogenesis of zebrafish. First by to factors transcriptional of binding the avoiding chromatin the process is histone deacetylase (HDAC) enzyme that remove an acetyl group from the histones condensing this of action the is mechanism regulation the development, of underline One unknown. theembryonic still is regulation this of during mechanism the formed however genes, different of organs expression the by temporally first and spatial regulated the of one is Heart Santiago, Chile aoaoi d Booí dl earlo Dpraet d Ceca Boóia, Universidad Biológicas, Ciencias de Departamento Desarrollo, del Biología de Laboratorio 247 1,2 mef2c, andJ. A. Ulloa this over expression is similar to the over expression observed when embryos 1 2 ilnim nttt fr udmna ad ple Biology, Applied and Fundamental for Institute Millennium n oe epeso o cric ee uh as such gene cardiac of expression over and vmhc hybridization, showing that showing hybridization, situ in Cardiovascular system in situ hybridization gata5, site copies the severe revealed a nonsense mutation in the open reading frame encoding for the positional cloning we were able to map the By chambers. heart both of contractility reduced severely to due flow blood of absence display screen for recessive lethal mutations that perturb cardiac contractility. site at amino acid position 195 (cMLC-1 In order to definethe thein rolecMLC-1 of contractility theof cMLC-1carboxy-terminus impaired regulation ofcardiaccontractility.carboxy-terminusthe we severely of mutatedrole a to crucial putativea lead phosphorylation suggesting mechanisms, cMLC-1 different of by truncation and ablation embryos, and mutant morphant cMLC-1 between similarities phenotypic the of despite that, demonstrates This in Interestingly,sarcomeres cardiomyocytes. in analysis showed that morpholino-mediated knock-down of cMLC-1 disrupts sarcomere assembly We recently isolated the zebrafish mutant light chain-1(cmlc-1)isonlypoorlyunderstood. disorder, precise cardiac the HCM, for inherited of One to common individuals. due mutations in generation sarcomeric young proteins. force Although myosin in ineffective light chains (MLCs) is most are death known HCM disease genes cardiac in implicated sudden mechanisms of pathophysiological the the cases the of is half over for (HCM) accounting cardiomyopathy Hypertrophic ofMedicine III,Department Heidelberg, University Hospital Germany C. Laufer,B.Meder, D.Hassel,H. A. Katus,W. Rottbauer Cardiomyocyte Contractility in Vivo Controls Chain-1 Light Myosin Essential Cardiac Carboxyl Terminusof the in Serine Single A Cardiovascular system controls cardiaccontractilityinvivo. terminus of cardiac MLC-1 in force generation and demonstrate that phosphorylation of cMLC-1 contractility in contrast, introduction of a phosphomimetic amino in acid (cMLC-1 cMLC-1 deficient morpholino-modified antisense oligonucleotide directed against the against directed oligonucleotide a antisense of morpholino-modified Injection defect. contractility the rescues cardiomyocytes mutant in cMLC-1 wild-type of in cMLC-1 truncated carboxy-terminally a and translation laz mutants. Our results indicate for the first time an essential role of the carboxy- lazy susan phenotype in 99% of injected wild-type embryos. Ultrastructural uat adoycts os o rsu te otatlt dfc. In defect. contractility the rescue not does cardiomyocytes mutant laz structural and functional role of particularly the particularly of role functional and structural vivo in cmlc-1 cardiac essential myosin light chain-1 (cmlc-1). Sequencing S195A lazy susan (laz gene leading to the premature termination of protein ). Strikingly, overexpression of this phosphorylation- lazy susan locus to a genomic interval including an mutant cardiomyocytes assemble normally.assemble cardiomyocytes mutant laz m647 ) in a large-scale ENU-mutagenesis uat ers Overexpression hearts. mutant laz S195D ) 248

fully restores cardiomyocyte Lazy susan mutant embryos cmlc-1 translational start translational cardiac essential cardiac 319

Posters Posters populations of stem cell-derived atrial cardiomyocytes. This knowledge may also impact our impact also treatment ofdegenerativeatrialdiseases. may knowledge This cardiomyocytes. atrial cell-derived stem of populations pure of differentiation for strategies novel to leading fate atrial promote that pathways signaling of suppressors of atrial differentiation. Our results suggest that Gremlin 2 controls the activity of atrial genes. Gremlin 2 induces atrial-specific thattranscription factors, whereas it blocks expression show also results Our development. treatment of ventriculardifferentiating mouse embryonic stem cells with restricts Gremlin 2 stimulates expression of and territory atrial the expands 2. Gremlin of lack to sensitive more Consistent is with this development idea, our results atrial show that affected, injection of are cells atrial ventricular both Although and development. cardiac in defects severe causes 2 Gremlin of inactivation Weheart. developing archthe pharyngeal to the next in mesenchyme expressed is 2 that found BMPThe antagonist development. cardiac proper for BMPrequired of is zone, activity are antagonists also necessary for cardiac development indicating that fine-tuning the strength, or confining the BMP secreted signaling, and BMP morphogenesis forward cardiac Besides on differentiation. cardiomyocytic effects pleiotropic have (BMP) Proteins Morphogenetic Bone Erkrankungen, Tübingen, Germany 1 320 Hatzopoulos A.K. cells toatrial Bone Morphogenetic Protein antagonist Gremlin 2 promotes differentiation of embryonic stem nvriy Nsvle U.S.A.; Nashville, University, Medicine, Department of Medicine and Department of Cell & Developmental Biology, Vanderbilt eeomna Booy Vnebl Uiest, ahil, ... and U.S.A.; Nashville, University, Vanderbilt Biology, Developmental Knapik iiin f adoaclr eiie Dprmn o Mdcn ad eatet f el & Cell of Department and Medicine of Department Medicine, Cardiovascular of Division 249 2

1 J Hu J. , 1, J Yan J. , 3 bradKrsUiestt üign Kii fr Herz-Kreislauf- für Klinik Tübingen, Eberhard-Karls-Universität 1, II Müller I.I. , 2,3 WH Heaton W.H. , gremlin gremlin 2 mRNA in zebrafish embryos Cardiovascular system 1 WD Wang W-D. , 2 iiin f Genetic of Division 2 ad . W. E. and , gremlin gremlin or whetherinsteadtheymayguidemotoraxonswhicharenecessary forformationofthePAV. HMS just as the PAV does. We are now testing whether Unc5b and DCC act in endothelial cells, PAV,the before shortly develops (RoP) neuron motor the primary along Rostral courses the and several for guidance axon in important neuronal be types. to In fact, known the is signaling HMS homologs is netrin Unc5 contacted and and/or by cells, axons DCC endothelial of through the guide primary might motor axons neurons. that In hypothesized particular, been often has It elsewhere. raising the question of whether these receptors act autonomously in endothelial cells, or perhaps down Knocking PAVduring HMS. the along lie which pioneers, muscle the by expressed is Netrin1a formation, the into the PAV. to rise give laterally and to to posteriorly Prior and grow anteriorly turn then and then muscle, superficial (HMS), myoseptum which horizontal sprouts, the secondary reach the they to until ~32hpf dorsally at grow rise gives PCV The (PCV). vein cardinal posterior Zebrafish trunk vasculature development begins with the formation of the dorsal aorta (DA) and receptors thatreceivethisNetrin1asignal. formation of the zebrafish parachordal vessel (PAV). Here we address the identity of the cellsvascular and in development. role In particular,important we an have shown play (Wilson molecules et al., guidance 2006) that Netrin1a axon is required that for the shown has evidence Recent e r ual t detect to unable are we However,DCC. and Unc5b through mediated likely PAVformation, promote to acts pioneers form sprouts secondary morphants, from the PCV,Netrin1a but they fail to turn at the HMS to form the PAV.as well as Thus, Netrin1a from the muscle morphants these In form. to fails also When we use morpholinos to knock down the known Netrin receptors Unc5b or DCC, the PAV PAV (Sulietal.,submitted). 1 A.H. Lim during formation vascular development vessel parachordal for required are DCC and Unc5b receptors Netrin The Cardiovascular system Dept of Neurobiology and Dept ofNeurobiology Anatomy, 1 , A. Suli or blocking muscle pioneer formation both prevent formation of the of formation prevent both formation pioneer muscle blocking or netrin1a 1 , D.Y. Li or unc5b 2 andC-B.Chien RA n eodr srus by sprouts secondary in mRNA dcc 2 Dept ofOncologicalSciences, University ofUtah, USA 1 250 hybridization, situ in 321

Posters Posters on an on tbx5b zebrafish both markers, valve cardiac Moreover,heart. vertebrate the shaping in step tbx5b anti-sense morpholino-treated embryos do not undergo looping morphogenesis, a critical the fin bud, suggesting a divergence in function between the two genes. Embryonicnot identical to heartthe expression pattern of tubes of transcription factor. highest homology in the DNA-binding T-domain, indicating that tbx5b retained its function as a the retaining while tbx5a, from diverged significantly is tbx5b sequence, acid amino on based expression of the cardiac marker, cardiac the of expression for tbx5a. Indeed, like tbx5a, tbx5b localizes to the cytoskeleton through direct contact with contact direct through cytoskeleton the actin-binding PDZ-LIM protein pdlim7. In summary,the the zebrafish paralogs to localizes tbx5b tbx5a, like Indeed, tbx5a. for laboratory our in demonstrated previously feature a compartment, cytoplasmic and nuclear the between shuttle to protein tbx5b of property the examined also we development, in roles their gene functioninrelationtoembryonicdevelopment. zebrafish of identification the Thus across species, and to the functional consequence of TBX5 mutations in human genetic disease. development in T-boxproteins of role conserved evolutionarily the to insight further lend may has identifiedanalysis similarcomparative andOur divergentproperties. rolesfunctional for and homology sequence conserved have 322 like that orthologs show studies Our disease. heart congenital in implicated heart similarly, two-chambered and zebrafish fins; the pectoral of of absence abnormalities and cardiac severe in results mutation (hst) cloning and characterization of a/b, tail no members, family the T-box Among genome. zebrafish the in reported been have duplication) zebrafish in members 19 T-boxof comprised factors Universitytranscription of family conserved evolutionarily an are (tbx) proteins Northwestern Center, Research Memorial Children’s Chicago, USA Pediatrics, of Department S. Park, T. Camarata,J.Topczewski, H-G.Simon Characterization oftbx5b,anovel paralog oftbx5a,duringzebrafish cardiacdevelopment 251 Tbx5 (mouse)/TBX5 (human), , and tbx2a/b, hst aaos ol rsl i cmon cric eet. y ncig down knocking By defects. cardiac compound in result would paralogs tbx5 -/- background, we found that a linear heart tube does form, but with attenuated with but form, does tube heart linear a that found we background, tbx5b expression is dynamic in the heart tube and dorsal eye, paralleling but and bmp4 represent examples of gene duplication. Here we present the present we Here duplication. gene of examples represent tbx3a/b . Gene paralogs (homologous genes that have evolved by evolved have that genes (homologous paralogs Gene . rerio Danio tbx5b, a novel paralog to zebrafish at 48hpf. In addition to a comparative assessment of assessment comparative a to addition In 48hpf. at cmlc2 . We tested the hypothesis that simultaneous loss of loss simultaneous that hypothesis the tested Weversican . affords an opportunity to examine the evolution of evolution the examine to opportunity an affords tbx5b tbx5a and tbx5b expression is critical for heart development. First, tbx5a. Surprisingly, however, in the mammalian four-chambered heart is heart four-chambered mammalian the in Tbx5 morphants demonstrate misregulation of misregulation demonstrate morphants tbx5b tbx5b in zebrafish development. These studies Cardiovascular system tbx5a. The n is mammalian its and tbx5a tbx5b lacks expression in tbx5a tbx5a and heartstrings heartstrings tbx5b latter dependsonVE-Cadfunction. formation” and “vessel assembly” participate as two distinct morphogenetic events and only the occurs during vessel that assembly. ECs We of behavior propose a migratory revised model for of ISV required formation, in is which “scaffold VE-Cad that suggest findings our Takentogether Cad, performed ECs fail to perform cellular rearrangements, which haveare required for proper vessel formation. we Analysis formation of junctional proteins and in vivo ISV time-lapse analyses indicate that in the during absence of VE- Initial sprouting of the (VE-Cad) ISV appears normal but they become disrupted during subsequent stages. VE-Cadherin of morpholino role knockdown experiments. VE-Cad the morphants labelling display multiple examine cardiovascular mosaic defects. To by Furthermore, outgrowth. tubes containinganextracellularlumen. sprout during multicellular are ISVs that shown have we proteins, junctional extensively of examination and experiments divide (ECs) cells study to paradigm a sprouting angiogenesis as in vivo. serve We embryo have previously zebrafish shown that the during ISV of formation endothelial (ISVs) vessels blood intersegmental The Dept. ofCell Biology, Biozentrum/Uni Basel, Switzerland H-G. Belting, Y. Blum,L.Herwig, A. Krudewig,E.EllertsdottirandM. Affolter VE-Cadherin isrequiredforendothelialrearrangements duringISVFormation Cardiovascular system 252 323

Posters Posters for myocardialmigrationduringcardiacfusion. demonstrate that Hand2, through modulation of Fn1 levels, creates an appropriate environment wild-type environment are able to integrate normally into the wild-type heart. Together, our data contrast, In normally. fuse from our cardiomyocytes wild-type with Consistent midline. the at together fuse hypothesis, we determined to through mosaic able analysis that the are cardiomyocytes of populations for the downmodulating by environment exposure of extracellular cardiomyocytes to the extracellular matrix controlscomponents like Fn1. Hand2 Indeed, that heterozygosity hypothesized we 324 of expression hand2 mutants. Moreover, gene overexpression of Through of upregulation observe Notably,we fusion. fundamental. cardiac for essential be may is in it regulated differentially are that for transcripts several identify we responsible profiling, components downstream the and markers, polarity of misplacement display aberrant fibronectin deposition. Given the dramatic show epithelium, an form not do cardiomyocytes the a polarity.In apicobasal reflect possessing may epithelium defect myocardial a fusion establish mutant to failure hand2 The fuse. never that cells of populations separated midline, the at tube heart a form to populations cardiomyocyte zebrafish the of causes development Mutation a striking process. cardiac heart fusion this phenotype. of for Instead of regulator critical exhibiting their the particularly essential proper migration be of an to withbilateral and appears that factor interact the transcription on they bHLH focus a we how Hand2, Here, together,understood. of poorly role come is regulated is populations fusion their these how and How environment, embryo. early the precursors cardiac of in populations separate two of fusion the from results heart vertebrate The Skirball Institute, NYUSchoolofMedicine, New York, USA Fibronectin of Control Z. Garavito-Aguilar andD. Yelon Through Fusion Cardiac for Levels Environment Proper a Ensures Hand2 fn1 mutants. 253 in fn1 mutants rescues aspects of the of aspects rescues mutants hand2 Based on this apparent reciprocal relationship between hand2 results in a cardiac phenotype similar to that mutant phenotype: notably,phenotype: mutant bilateral hand2 the hand2 hand2 cardiac phenotype, knowing mutant cardiomyocytes in a in cardiomyocytes mutant hand2 Cardiovascular system mutants maintain two maintain mutants hand2 mutant environment prevents hand2 and fibronectin1 (fn1) fibronectin1 mutants and mutants hand2 mutants, hand2 fn1 function, gene hand2 in Cayuela 2 Alcaraz-Pérez F. The roleoftelomerase ininnateimmunityandviral resistanceofzebrafish Hematopoiesis andimmunology 1 r w cmoet o tlmrs wt mrhlns eutd n mard eeomn and developmentanextracurricularroleoftelomeraseisrequiredforimmunity. impaired in resulted morpholinos withfunctions of immune cells, which led to increased viral susceptibility. telomerase These results indicate that of components one of two Knockdown (SVCV). or virus carp of viremia spring to resistance and immunity innate in of life. In this absent. study, we still have used zebrafish embryosis to determine the roleimmunity played by telomerase adaptive when time Therefore, innate a immunity plays an at essential role in environment the survival of aquatic this specie at the the early stages of load microbial fluorescence- of organslymphoid labeled cells. immune and high the to exposed is zebrafish the hatching, After observation easy for allowing thus ex-utero, rapidly develop and transparent are fish thethis of embryos and immunity.The developmental studying cells for model vertebrate immune excellent innate in telomerase an is ) rerio (Danio zebrafish of The understood. poorly remain infection to cells role these of response the However, aging. cellular and stress, the losses of replicative capacity and mitigate function caused to by believed chronic is antigenic cells stimulation, immune oxidative these in Telomeraseactivation challenge. an immunological to response in telomerase up-regulate transiently to able are lymphocytes B and activity,T telomerase no or little show which cells, somatic normal most integrity.to telomere contrast In enzyme can trigger cellular DNA damage responses in both the presence and absence of altered this of loss The aging. cellular retarding and fusion and recombination from ends chromosome protecting chromosomes, linear of ends Telomerasethe DNAonto telomere transcribes reverse Departement ofCellularDepartement Biology, University ofMurcia, Espinardo(Murcia), Spain Spain;Arrixaca’,(Murcia), la Palmar de El ‘Virgen Hospital Surgery,University of Department 1 1,2 M A López-Muñoz A. M. , 2 M Anchelin M. , 1 M Moreno M. , 254 1 V Mulero V. , 2 M L. M. , 325

Posters Posters 326 like differentiation, intrathymic lymphoid of characteristic genes marker other with together thymus the in expressed is it stages their and cell mass (ICM), thymus the first hematopoietic structure detectable in the embryo. By the 54 hpf and at later to key factors is homing these of One factors. transcription different of network the complex a by directed is differentiation comprising development, Lymphocyte and stromalcellsthatisrequiredtoestablishaself-tolerantrepertoireofTcells. mature T cells. The thymus provides a special microenvironment for the interaction of lymphoid first detectable at 48 hpf. It is colonized by lymphocyte progenitor cells, which differentiate into In zebrafish, the thymic anlage develops from the pharyngeal endoderm as a bilateral structure, Max-Planck-Institute ofImmunobiology, Freiburg, Germany I. Hess,M.Schorpp,D.Diekhoff,T. Boehm Watching thymopoiesis instabletransgenic zebrafish embryos rgntr bt hi tyu hmn i ipie, niaig ht hs ceoie pa an play chemokines these that indicating impaired, important roleinthymushoming. is homing thymus their but progenitors cell T cells subsequent and development. lymphoid Wethymus also the show, and into that ccl25a/cxcl12a stromal morphants prothymocytes show normal of numbers of of lymphoid immigration interaction normal for the important is and TECs of differentiation the that indicating antisense against gene-specific directed (MOs) with interference functional Of special interest to us is the process of by thymus homing. We show that injection of morpholinos latter morpholinos. the thymopoiesis, mutations various affecting introducing by investigated is former The vertebrates. in homing cell T and development thymus influencing factors the characterize to lines transgenic these use Wealso anlage. anlage. thymic the the to Using tissue) hematopoietic (caudal CHT the and AGM the mesonephros) ICM, (aorta-gonad the from cells hematopoietic of pattern migratory the of characteristics spatial generated we vivo, the Using imaging. time-lapse confocal long-term in thymopoiesis of development of the thymic epithelial anlage and the homing of prothymocytes to the thymus in the follow process to us allow lines These the BAC-DNA. modified using lines transgenic study foxn1::mCherry to order In (TEC) differentiation.Inzebrafishitsexpressionisdetectablefrom48hpfonwards. 255 ie w dsrb te epr-pta dvlpet f h thymic the of development temporo-spatial the describe we line, foxn1::mCherry . ikaros In zebrafish, . Another important factor is factor important Another rag1. ed t aern mgain f eaooei poeio cells, progenitor hematopoietic of migration aberrant to leads foxn1 ikaros expression is first detected at 24 hpf in the intermediate line, we describe the temporo- the describe we line, ikaros::eGFP Hematopoiesis andimmunology , a regulator of thymic epithelial cell epithelial thymic of regulator a foxn1, and ikaros::eGFP H.B. Taylor dependent kinase MAP p38 is model injury embryo zebrafish a in recruitment neutrophil enhanced LPS Hematopoiesis andimmunology odnEibrh UK; London/Edinburgh, 1 activation. TLR4 of independent process inflammatory acute the and recruitment leukocyte of modulators allows in vivo investigation of the mechanisms of LPS recognition in fish and evaluation of other LPS recognition and TLR4 signalling in fish are different from the mammalian system. This model suggest that findings recent complement study this in presented results The unknown. relatively are challenges PAMP other and LPS to responses transcriptional and functional Fish TNFb cytokine. and TNFa of expression increased with manner dose-dependent a in stimulation LPS to respond did cells derived marrow kidney whole adult but LPS with challenged when activation differentially were zTLR4b p38 and of zTLR4a activation zTLR4b. expressed in embryo the and and adult tissue. Zebrafish myelomonocytes via zTLR4a did incompletely not display morphological of wound independent tail remain was a but to thisprocess. recruitment MAPK recruitment image neutrophil and of study enhance to to mechanisms found model was injury the LPS embryo zebrafish where a use infection we Here and understood. injury local response inflammatory complex to the of component important an is leukocytes of Recruitment Biology, ImperialCollege London, London, UK Division of Cell and Molecular Biology/CIR, Imperial College London/University of Edinburgh, of London/UniversityCollege Biology/CIR,Imperial Molecular and Cell of Division 1 , S.B.Brown 2 , J.R.Lamb 2 I, nvriy f dnug, UK; Edinburgh, of University CIR, 3 , M.J.Dallman 3 3 iiin f el n Molecular and Cell of Division 256 327

Posters Posters also showed that zebrafish embryos use both MyD88-dependent and -independent signalling pathways andidentifiednovelimmuneresponsetargets ofTLR5andMyD88signaling. -independent and MyD88-dependent both use embryos zebrafish that showed also flagellin, which is the first demonstration of a conserved TLR ligand specificity in this model. We knockdown analysis we showed that study zebrafish TLR5 is profiling required for transcriptome the recognition of Our transcription. antisense revealed specific of regulation of level high Toll-like receptor a 5 and and downstream databases, signalling components.EST Using and transcript current the in present not products splice novel of expression switching, isoform insights that microarray analysis could not have achieved, such as infection-dependent transcript provided sequencing deep of nature unbiased the genes, of groups functional similar identified sequencing deep and microarrays Although annotation. genome vertebrate support will which of characterization functional future the of markers, infection set and a large genes response immune of novel identification the to led study our Furthermore, mediators. inflammatory and induction of genes encoding cell surface receptors, clear signalling intermediates, transcription showed factors mutant O-antigen LPS avirulent conservation the with host and responses detected strain in lethal other vertebrate the models and both human cells, with including infection to model sequencing two with deep infection to novel strains that response elicit either a lethal infection or an immune and attenuated response. The transcriptional response innate microarrays embryonic the used of analysis We ontology gene infection. and study profiling transcriptome time-course bacterial detailed a perform to technology to response the of zebrafish embryo represents a useful Due to the clear separation of innate immunity from adaptive responses, the externally developing Research Center, Szeged, Hungary 1 328 during A. H.Meijer signalling receptor zebrafish embryos toll.like and responses Transcriptome Institute of Biology,University,Netherlands;Leidenof Leiden,The Institute 257 1 , O.W. Stockhammer 1 , A. Zakrzewska in vivo model for identification of innate host determinants 1 Hematopoiesis andimmunology , Z.Hegedus 2 2 , H.P. Spaink ZenonBio Ltd. and Biological and Ltd.ZenonBio neto of infection salmonella 1 Salmonella Salmonella the resolution of inflammation. This delay in resolution was not observed in notobserved was inresolution delay delay significantly This to found inflammation. was of DMOG resolution with the pathway signalling hypoxic the of Activation techniques. microscopy fluorescent using observed be can that response inflammatory neutrophilic limiting 1 resolution. of the hypoxic signalling apparatus dissection during molecular our in aid to model zebrafish the of potential the highlight studies These on effects their neutrophil behaviourandtheresolutionofinflammation. of observe to forms neutrophils mutant in them These expressing pathway. specifically hypoxia before characterisation the of readout a as acts line or upregulate PHD3 (prolyl hydroxylase successfully 3) is a known these downstream target of that HIF1α and thus indicated the have downregulate the HIF studies pathway respectively, by microinjection into a Initial cloned. been have hif1α Using molecular biology techniques, dominant active and dominant negative forms of zebrafish TILLING mutants. the not in or DMOG was by pathway hypoxia neutrophils the of upregulation of the after differ index significantly to found meandering and velocity The approach. timelapse microscopy confocal disk spinning novel a using investigated was behaviour Neutrophil embryos. mutant Using zebrafish to investigate the molecular control of hypoxic signalling during signalling ofhypoxic control molecular the P. investigate Elks to inflammation zebrafish Using Hematopoiesis andimmunology UK; (DMOG) or by outcrossing to outcrossing by or (DMOG) dimethyloxaloylglycine of addition the by embryos these in upregulated was pathway hypoxia gene specific neutrophil the of expression the by driven is (GFP) protein fluorescent green which in line transgenic a using zebrafish the in visualised were Neutrophils pathway canbeactivatedbyinflammatorystimuliindependentlyofhypoxia. and inactivated are hydroxylases However,targets. downstream HIF of the activation the and transcription allowing stabilised is HIF hypoxia, In degradation. by for phosphorylated is targeted HIF and normoxia, hydroxylases In manner. dependent oxygen neutrophil an in prolonging protein (HIF) stimulus factor is aprofound the to lead may Hypoxia functional lifespan. The hypoxia mechanismsgenetic pathway regulates the targets. stability of the these hypoxia inducible therapeutic of of knowledge greater identification a and understood, poorly remain regulated is function neutrophil which by mechanisms genetic The untreatable. largely remain and world, developed the in problem increasing an are inflammation neutrophilic of Diseases MRC Centre for Developmental and Biomedical Genetics, University of Sheffield Medical School, 2 Academic Medicine, UnitofRespiratory University ofSheffieldMedicalSchool, UK 1,2 , F. vanEeden 1,2 , M.K.BWhyte or vhl null TILLING mutants. Tail transection induces a self- a induces Tailtransection mutants. TILLING null fih 2 , S.Walmsley in vivo physiological processes such as inflammation 2 , S.Renshaw 1,2 258 phd3:GFP transgenic line. myeloperoxidaseThe . eur further require hif1α Tg(phd3:GFP) vhl -/- or 329 fih -/-

Posters Posters ae f aiuain and manipulation of ease nvriy Hlfx Canada, Halifax, University, stabilizing agent, ketotifen. Similarly, infection with heat-inactivated complete maturation where maturation complete carboxypeptidase A5 (cpa5) an RI signaling. We are characterizing an analogous an characterizing Weare signaling. IgE/FcεRI MCs. Intraperitoneal injection of compound 48/80, a MC 330 responded DNP-BSA of injection by followed IgE anti-DNP mouse with al., et (Dobson first the were environment. These anatomic locations position them to play 1 S. Da’as Zebrafish MastCellsDemonstrate Conserved Innateand Adaptive ImmuneResponses the presence of an IgE-like receptor on zebrafish MCs. Functionally, zebrafish MCs sensitized MCs zebrafish Functionally, MCs. zebrafish on receptor IgE-like an of presence the localization of zebrafish receptor. homologues of IgE IgE receptor affinity subunits with high expression of human co- showed subsequently we embryos, day 7 the on hybridization RNAsitu mount in whole Using of subunit gamma the targets specifically that antibody we have demonstrated positive immunostaining in zebrafish MCs for a rabbit-derived polyclonal Mast cells (MCs) are multifunctional immune cells derived from derived cells immune multifunctional are (MCs) cells Mast Farber CancerInstitute, Boston, USA and elevated plasma tryptase plasma elevated and diseases. Toll-like well-conserved through mediated be may receptors. which pathogens, to responses immune innate in participate MCs suggest results These interestingly,recruitment. and eosinophil levels tryptase plasma increased degranulation, MC zebrafish in resulted zymosan, constituent, wall with a goal of identifying of goal a with and leukotriene production. Our studies reveal conserved MC function in zebrafish innate zebrafish in zebrafish function the establishing MC effectively conserved reponses, immune reveal adaptive and studies Our production. leukotriene and evaluating vertebrate MC activity. Ultimately, we will be able to exploit the zebrafish system zebrafish the exploit activity.to MC able vertebrate Ultimately,be evaluating will we new model system for studying infection and immunity and infection studying for system model new innate both eliciting IWK HealthCentre, of Departments platform for high-throughput screening of potential of screening high-throughput for platform vivo in 259 1 , E.MTeh

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Nova Scotia, Canada, 1 S. Da’as, Notch signalingisrequiredformastcelldevelopment inthezebrafish Hematopoiesis andimmunology expression. Similarly, wild type zebrafish embryos treated with Compound E, (Cpd E),a previously We (Cpd phenotype. E, similar a identified show Compound signaling, with Notch inhibits treated that embryos inhibitor zebrafish γ-secretase type wild Similarly, expression. 5 mutant, signaling Notch zebrafish the that shown now have marker. We MC-specific zebrafish a as (cpa5) 5 carboxypeptidase A regulation of significance transcriptional MC vertebrate examine al., et (Dobson equivalents MC zebrafish decisions. identify to first fate the were We cell MCs. lineage including cells, and hematopoietic of number a induction in expressed are ligands their cell and receptors Notch stem hematopoietic been regulating has and in metazoans all implicated in be conserved highly to is remain pathway vertebrates signaling Notch in The development elucidated. (MC) cell mast regulating pathways molecular The Center, New York, USA inhibition mediated through mediated inhibition and zebrafish embryos treated at 50 μM Cpd E show decreased novel therapeuticstrategiesinMCdiseases. an as zebrafish the use to opportunity the and development MC in line are also being undertaken. These studies promise key insight into the role of Notch signaling to test Notch pathway inhibitors in this condition. Parallel studies in a human mastocytosis cell employed be will mastocytosis, systemic condition, preleukemic the in found mutation D816V MCs. Moreover, a second transgenic line generated in the laboratory expressing the human in the laboratory, which will enable the generated tracking of been Notch signaling in has developing and migrating line zebrafish reporter (GFP) protein fluorescent green Notch A Notch. of bomb and Cpd E treated) are underway to (mind determine the absent specific signaling cascadeis downstream pathway Notch the which in embryos in factors transcription other and gata-2 experiments signaling Reciprocal Notch zebrafish. overexpressing the that in development suggest MC for data necessary Our pathway critical development. a is lineage erythroid and myeloid both in pu.1, W Hat Cnr, eatet of Departments Centre, Health IWK expression, suggesting a particular sensitivity of the MC lineage to Notch pathway Notch to lineage MC the of sensitivity particular a suggesting expression, gata1 , and , gata1 1 L.Rygier, pu.1 and notch mRNA in wild type embryos and rescue experiments overexpressing , suggesting a broader dose dependent role for Notch pathway signaling pathway Notch for role dependent dose broader a suggesting mpo, 2,3 gata2 as essential transcription factors for early MC development. Wild type A. Ferrando 4 Institute for Cancer Genetics, Columbia University Irving Cancer Research . At 75 μM Cpd E, we observed severe reductions in in reductions severe observed we E, Cpd μM 75 At . gata2 4 , J.N.Berman 2 ilg and Biology 1,2,3 mind bomb, mind Tee tde dmntae the demonstrated studies These vivo. in 3 eitis Dlose nvriy Halifax, University, Dalhousie Pediatrics, expression, gata2 but wild type displays profound loss of loss profound displays in vivoidentifying in for model 260 2008) and 2008) Blood , gata2 pu.1, CKIT pu.1 cpa- 331

Posters Posters 332 craniofacial including associated withU6snRNA inthespliceosome.PossiblefunctionsofLsm8willbediscussed. abnormality proteins Sm-like of member a encodes Lsm8 gene. Lsm8 developmental the in mutation nonsense a identified apparent no structures, have but Homozygotic exhibit WW18/10. a mutants, reduction mutants of the immature of T-cellsone WW18/10 in on the report embryonic we thymus. 10 Here Positional identified cloning lines. have we 41 writing, among this mutations of time the At thymus. the in T-cells immature expressing rag1- of accumulation abnormal with lines zebrafish ENU-mutagenized of panel a established of diseases. However, establishment the autoimmune relevant the and molecular syndromes immunodeficiency pathways controlling for are for crucial not is necessary development fully thymus understood. is We have thymus previously the in and lymphocyte T for mechanisms the Understanding vertebrates. in systems immune acquired lymphocytes T of Lsm8 inthymus development Proper for requirement Max-Planck-Institute ofImmunobiology, Freiburg, aspecific Germany N. Iwanami, M.Schorpp,andT. Boehm identifies zebrafish development in ENU-mutagenesis 261 Hematopoiesis andimmunology o J e a (07 Dsoey f utpe eadfni lk hmlge i tlot fish. teleost in homologues like beta-defensin multiple Immunol 44(4) : 638-47 of Discovery (2007) al et J. Zou 318(5855) :1418-23 Candille SI et al (2007) A valuable diseasemodelforthestudyofβ-defensinsinthisresearch group. mammalian and intrinsic both of function the investigate to model zebrafish the of use novel the describes work this Overall, of knockdown Mc1r. morpholino the after and before both dispersal pigment melanophore on have cultured zebrafish melanophores to directly observe the effects that a panel of human between interactions appear darker in colour. This process can be observed under the microscope and is controlled by to background adapt by dispersing the pigment (melanin) within their melanophores in order to Human In addition, the zebrafish system was exploited to further define theanti-microbial assays. reported interactions between RT-PCR,quantitative to and addition hybridisations in in-situ knockdowns, morpholino function expression, their analysing by and inducibility in both embryos and adult fish. This has been achieved using a combination of genes these to functionality assign zebrafish to three aims and of 2007) reporting initial the on builds work This an provides This beta-defensins chemoattractmacrophagesisasyetunknown. cells. which T through receptor The and systems. immune adaptive macrophages and innate the between cells, link important dendritic immature as such cells immune vertebrate the Although innate immunecomponentsinavarietyofspeciesthroughouttheanimalandplantkingdoms. The Western GeneralHospital, Crewe Road South, Edinburgh, UK N. L.Reynolds Characterizing zebrafish defensins Hematopoiesis andimmunology 1 optl Cee od ot, dnug, UK Edinburgh, South, Road Crewe Hospital, London,UK Institute for Genetics and Moecular Medicine, MRC Human Genetics Unit, Western General WesternUnit, Genetics Human MRC Medicine, Moecular and Genetics for Institute -defensins make up one of the largest groups of antimicrobial peptides and are important are and peptides antimicrobial of groups largest the of one up make β-defensins -defensin 3 (HBD3) and the Mc1r receptor (Candille et al, 2007). Zebrafish are able are Zebrafish 2007). al, et (Candille receptor Mc1r the and (HBD3) 3 β-defensin dfnis lo ae eldcmne rls n h ceotrcin f adaptive of chemoattraction the in roles well-documented have also β-defensins 3 Institute for Genetics and Molecular Medicine,Molecular and Genetics Researchfor Cancer Centre,Institute Edinburgh -defensins were first identified for their broad-spectrum anti-microbial activities, anti-microbial broad-spectrum their for identified first were β-defensins 1 , D.Macmillan -MSH and Mc1r. This system has been exploited using freshly prepared, freshly using exploited been Mc1r.has and system α-MSH This β-defensin mutation causes black coat color in domestic dogs. 2 , C. A. Semple -defensins. This also represents the development of a novel, a of development the represents also This β-defensins. 1 , E.E.Patton 2 eatet f hmsr, nvriy College University Chemistry, of Department 3 , J.R.Dorin dfni-ie ee (o e al, et (Zou genes β-defensin-like 262 1 β-defensins Science vitro in 333 Mol

Posters Posters G. Lutfalla University, Denmark; 334 IFNSin induced virus for the D. Aggad receptors constitute zebrafish chains of CRFB combinations Specific 1 a commonshortchain,associatedtospecificlongchain. dimer.include different complexes a receptor on Both rely IFNs II receptor,class two the while described previously the through signal IFNs I class two the receptor; same the to bind IFNs all not that found we experiments, function of loss and gain- CRFB potency.Using marginal only has IFNφ4 while effective most the appearing IFNφ1 with challenge, viral a these to larvae zebrafish that found classImember: we a proteins, recombinant of of resistance the increase and viperin as such genes antiviral of expression the induce IFNs four injection encoding or sequence, studies afourth overexpression By chromosome IFNφ4. adifferent on identified have cluster,we chromosomic single a on sit genes three these While II. class to belong IFNφ3 and of conserved cystein pairs. Zebrafish IFNφ1 belongs to class I; the more recently described IFNφ2 viro-induced interferons has expanded, with two fish of number distinct the then, Since classes CRFB5. and CRFB1 definedof dimer a accordingas IFNL1) or to identified IFN1 as known thealso previously number We have genome. zebrafish the in zebrafish identified interferon viro-induced the first the of in receptor the functionally exist members proteins, 13 transmembrane least CRFB at of which of heterodimers are cytokines) IL-10-related and (interferons but the identity of their receptors is difficult to predict. The receptors of class II helical cytokines The number of fish proteins identified as cytokines by sequence homology has grown tremendously, CNRS, Montpellier, France; 263 1 , M. Mazel M. , 1 , J-P. Levraud 1 , P., Boudinot 5 New Jersey MedicalSchool, Newark, USA 2 2 Institut Pasteur, Paris, France; 3 , O. J. Hamming J. O. , 4 Hematopoiesis andimmunology , R. Hartmann R. , 3 INRA, Jouy-en-Josas, France; 4 , S. Kotenko S. , 5 , P., Herbomel (IFNφ1, 4 Aarhus 2 , in theCHTniche. residing cells progenitor haematopoietic from output differentiation myeloid and erythroid the regulating in TIF1γ for role novel a uncovered thus Wehave niche. haematopoietic this in fate cell myeloid restrict to required is CHT,TIF1γ the suggesting in markers myeloid of expression arising the absence of TIF1γ. Thus, TIF1γ is required for the survival blood cells was observed, suggesting that these cells are unable to complete their maturation in or differentiation of the erythroid cells erythroid markersarealsoexpressedintheCHTofmonmutants,norecoverycirculating red CHT.the Although in found are markers myeloid and erythroid expressing cells 4dpf, At cells. primitive circulating of interference the without studied be can haematopoiesis of waves later Because survival. cell red primitive for essential Trim33),or (TIF1γ factor 1γ a factor intermediate transcriptional ligase, ubiquitin domain RING mutant zebrafish the of use made about known is little how the differentiation present, potential of HSCs in the different niches is regulated. At Toequivalent). address this, we marrow bone (the kidney 2dpf the to and From thymus the aorta. to dorsal finally and progenitors, erythromyeloid the arising locally with co-exist of they where equivalent), wall ventral the with onwards in the association zebrafish, they migrate to the caudal haematopoietic tissue (CHT, in the foetal liver arise wave throughout HSCs definitive haematopoiesis adulthood. maintain a which by (HSCs), cells followed stem haematopoietic is generates that haematopoiesis primitive development, vertebrate During haematopoietic from output MHU, lineage WIMM, University ofOxford, UK myeloid R. MonteiroandPatient and erythroid progenitors the regulates TIF1gamma Hematopoiesis andimmunology de novo in the CHT. Surprisingly, TIF1γ-deficient embryos showed a dramatic increase in moonshine ( mon). mutants are devoid of primitive red cells, red primitive of devoid are mutants mon harbours a mutation in the nuclear the in mutation a harbours mon 264 335

Posters Posters determine howthesepathwaysdiffertothosecontrolling‘steady-state’ haematopoiesis. to and process this drive that pathways(s) genetic the dissect further to potential have also They context. animal whole a within myelopoiesis demand-driven observe directly to means novel a as well as the capacity of different purified TLR ligands to replicate it. These studies will provide contribution of TLR signalling mediated through the different TLR adaptors towards this response the identity. assessing precursor are myeloid We definitive early an suggesting larvae, depleted the dependent on HSC emergence within the of same region, as evidenced are by their absence expression in Runx1- clusters their leukocyte These by vein. highlighted cardinal also posterior and aorta dorsal the by bordered niche phagocytic haematopoietic of the within observed is infiltration leukocytes myeloid of expansion rapid massive a leukocytes, the by evidenced response immune of delivery following GFP-labelled days 2 Approximately challenge. bacterial live a and stimulation ligand TLR to response in myelopoiesis ‘demand-driven’ demonstrate larvae zebrafish that reveal we 336 transgenic within imaging live Using molecules necessaryfortransducingTLR-mediatedsignalling. to reporter lines, we have ability previously demonstrated that zebrafish their immune cells express the for adaptor characterised well are regulate immune responses of and mature immune cell subsets. pathogens Using live imaging within of transgenic components invariant conserved highly recognise that receptor recognition pattern of class ancient an are TLRs leukocytes infection. during myeloid of production the receptor Toll-liketowards for differentiation HSC role skewing in a signalling suggested (TLR) have studies Recent understood. incompletely are this or ‘emergency’ via stocks cell immune depleted ‘demand-driven’ regenerate haematopoietic activity.to However, potential the mechanism(s) through which it the achieves has host the that full the generate for rapid replenishment is to essential. During the stressed state of infection potential it has long been known lineage in are crucial for host protection. These innate immune reduction cells have a finite life span so a mechanism step-wise a that system immune innate the of leukocytes myeloid including lineages, blood of complement undergo (HSCs) haematopoietic haematopoiesis, cells state’ ‘steady stem normal During the host. for the critical is of challenge wellbeing pathogenic ultimate a to system immune innate the of response rapid The of Auckland, New Zealand University Sciences,The Pathology,Medical and of Medicine School Molecular of Department and Crosier K. MacDonald, P.A. T.Purea, Crosier T.Storm, Lam, E. Chien, A. Flores, Vega M. Hall, C. Live imagingdemand-driven haematopoiesisduringinfection 265 Salmonella typhimurium into the hindbrain of 50 hpf embryos, and a robust innate and Tg(lyz:EGFP/DsRED2) rngn ad hi pouto is production their and transgene runx1P2:EGFP Hematopoiesis andimmunology eotr lines reporter Tg(mpx:GFP) lyz:DsRED2-expressing 1 the idea that the TLR4 receptor complex for LPS recognition arose after the divergence of fish of and tetrapods. divergence the after arose recognition LPS for complex receptor TLR4 the that idea the this receptor of in most absence fish species, the explains the resistance with of fish together to endotoxiczebrafish, shock andthe supports in signaling TLR of regulator negative a as TLR4 of negatively regulated the MyD88-dependent signaling pathway. orthologs TLR4 We zebrafish surprisingly, the believe and, that fish in the manner identification MyD88-independent and a TLR4- via signaled LPS that show results Our orthologs. TLR4 two possesses which Cypriniformes) , rerio ( Danio zebrafish the (iii) and ortholog; TLR4 a lacks Tetraodontiformes)that nigroviridis, in which the presence of a TLR4 ortholog is unknown; (ii) the spotted green pufferfish (Tetraodon (i) the gilthead seabream (Sparus , aurata Perciformes), an immunological-tractable teleost model studied: were models fish bony different three and developed was embryos zebrafish whole in the role of fish TLRs in LPS recognition, a dual-luciferase reporter assay to study NF-κB activation mammals. of that from different be may fish in recognition LPS of mechanism on light throw To the that hypothesize to us led available, databases MD-2 EST and i.e. genomes fish TLR4, the all via in CD14, activation and LPS for are molecules co-stimulatory amphibians, essential the and of lack fish the and notably most vertebrates, lower that resistant to the toxic effect of LPS. Furthermore, the lack of a TLR4 ortholog in some established fish species been long has It ofSurgery,Department University Hospital dela “Virgen Arrixaca” Murcia, Spain M. P. Sepulcre New insightsintotheevolution oflipopolyccharide (LPS)recognitionandsignaling Hematopoiesis andimmunology V. Mulero eatet f el ilg, aut o Booy Uiest o Mri, Spain Murcia, of University Biology, of Faculty Biology, Cell of Department 1* 1 , F. Alcaraz-Ruiz 2 , A. López-Muñoz 1 , F. J. Roca 1 , M. L. Cayuela 266 2 , J. Meseguer 2 eerh Unit, Research 1 and 337

Posters Posters 338 the of increase weak a in resultedmRNAof levels with (PAMPs) zebrafish patterns adult of molecular Injection low. pathogen-associated very several was liver the in expression their although tissues, adult in expression ubiquitous an showed transcripts both hand, other the On development. of and md1 transferred, while that of and RT-PCR real-time by MD1 and their ability to In physically RP105 interact. The results revealed zebrafish, that the transcript of of signaling. profiles LPS the expression in the in involved analyzed not present we study, are this TLR4s all zebrafish are that shown orthologs recently MD1 have we and although RP105 TLR4b TLR4a, However, cells. CD14, what may explain the inability and of low MD2 concentration (ng/ml) of LPS orthologs to stimulate lack fishInterestingly,fish immune LPS. for receptor decoy a as acts which complex MD2/CD14 receptor complex. This response is functional negatively regulated by the RP105/MD1 receptor Mammalian and immune cells are able to sense bacterial lipopolysaccharide (LPS) by mean of TLR4/ molecular signaling: (TLR) ofCellDepartment Biology andHistology, Faculty ofBiology, University of Murcia, Spain receptor M. P. Sepulcre,S.Candel,J.Meseguer, Toll-like V. Mulero of characterization ofzebrafish MD1andRP105 regulation Negative that zebrafish MD1 was unable to interact with RP105 and TLR4 orthologs. Studies arein orthologs. and TLR4 TLR4, RP105andMD1inthezebrafish. RP105 by played roles with functional the illuminate to down tointeract knock morpholino-mediated using progress unable was MD1 zebrafish that found However, we zebrafish. the in pathways signaling RP105 and TLR4 between interaction functional PAMPs,a different suggesting to response in level transcriptional the at up-regulated zebrafish that indicate may results These LPS. by 267 may not require each other to perform their functions at least in the first hours first the in least at functions their perform to other each require not may rp105 tlr4a, and tlr4b rp105 was not detected until 9 hpf. These results suggest that zebrafish in the injection site, while that of that while site, injection the in rp105 orthologs and orthologs tlr4 Hematopoiesis andimmunology are simultaneously are rp105 was only induced only was md1 md1 was maternally 2 M. A. López-Muñoz Molecular andfunctionalcharacterization ofzebrafish IFNg1-1 Hematopoiesis andimmunology 1 not atrueIFNγortholog. is IFNγ1-1 zebrafish that propose we PF00714), conserved no. the accession (Pfam of domain lack superfamily IFNγ the and vertebrates higher of IFN γ with IFNγ1-1 sequence zebrafish low of the homology with together results, these account into Taking infection. bacterial or viral either to resistance their affect not did IFNγ1-1 recombinant with fish of importantly,injection More cells. these in IFNγ1-2 by exerted effects powerful the contrasting Mx, of expression the induce to unable was IFNγ1-1 with cells ZF4 of stimulation addition, In IFNγ1-1. of expression the induce to failed infection viral and bacterial that show results The models. infection SVCV) virus, its carp of and viremia (spring IFNγ1-1 viral and zebrafish iniae) (Streptococcus ofrecombinant bacterial using activity the production biological here report We fish. in understood poorly is infections viral and bacterial in IFNγ1-1 of relevance vivo However,in responses. the are expressed in immune cells of teleost genes fish and are potentially implicatedTwo in B-IFNγ and infections. T- lymphocyte bacterial intracellular and viral to responses immune and adaptive innate the of regulation the in involved cytokine important an is (IFNγ) gamma Interferon eatet f el ilg ad itlg, aut o Booy Uiest o Mri, Spain Murcia, of University Biology, of FacultyResearch Unit, Histology, ofSurgery, Department University Hospital dela Virgen Arrixaca, Spain and Biology Cell of Department 1 , M.P. Sepulcre 1 , F. Alcaraz-Pérez 1,2 , J.Meseguer 268 1 , V. Mulero 1 339

Posters Posters F. Cotelli 1 340 Bresciani E. Zebrafish NumbandNumblikeareinvolved inprimitive erythrocytes differentiation f ub n Nmlk i eyhoye dtriain n dfeetain uig primitive during differentiation and determination erythrocytes in hematopoiesis. Numblike and Numb of first the provide Takingresults venosus. together,our sinus the within present is cells red of amount little a only 72hpf and 48hpf at and reduced strongly of fluorescence overall the development, of stage this at that, morpholino in the transgenic the line : Tg(gata1 injected DsRed). Injected we embryos defects at erythropoietic about 28-30hpf the showed into insight gain to Furthermore, detectable. were cells blood red differentiated few only that observed we and morphants 48hpf in staining erythrocytes in nb/nbl time earliest the primitive differentiated of presence the assess to order circulationIn from that detected. point be can starting reduced severely embryos or observedabsent are we cells blood Numblike circulating which and in Numb of knock-down (MO) oligonucleotide antisense morpholino Using embryo. the of half anterior the to restricted become expression their 24hpf by organogenesis and to stages stage cleavage first the from embryo whole the in expressed are differentiation. neuronal maintaining and specifying in functions redundant play (Nbl) Numblike homologue its and (Nb) Numb that suggest evidence of lines several mice, In Numb. protein adaptor conserved evolutionary the is and renewal self HSCs included both differentiation lineages processes blood developmental various regulating in in determination implicated fate system cell regulatory conserved evolutionary an is signaling Notch The blood lineages. In mature definitive to rise give to able while (HSCs) types. Cells Stem Hematopoietic term macrophages long provides hematopoiesis primitive cell some blood only and analogous erythrocytes predominantly of produces formation the drive Vertebrates blood which cells formation consists in two successive waves. The programsprimitive hematopoiesis genetic conserved Zebrafish hematopoiesis has been shown to be very similar to that in mammals with evolutionary Numb its interactionwithNotch. provided evidences that Numb can modulate the specification of primitive erythrocytes through both Nb and Nbl are dispensable for hematopoiesis in adult mice but recent could play a proteins role Nbl and also Nb that in possibility the the rise embryonic findings and these Hence, adult models. human hematopoietic and system. mouse It has been proposed that both in tissues hematopoietic adult and sac yolk the including embryos mouse of tissues the of Dipartimento diBiologia,Dipartimento Università degliStudidiMilano, Italy; 269 and 1 Numblike homologues have been identified and cloned also in zebrafish where they 1 , S. Confalonieri S. morphants we analyzed the erythroid hemoglobin content by o-dianisidine 2 , S. Cimbro S. , and vitro in The expression of expression The 1 , E. Foglia E. , . One known inhibitor of Notch activity Notch of inhibitor known One vivo. in Hematopoiesis andimmunology 1 , S. Carra S. , and Nb in vivo in nb/nbl 1 , C. Lora Lamia Lora C. , has been detected in most in detected been has Nbl 2 IFOM, Milano, Italy evidence of an involvement an of evidence morphants start to appear to start morphants in vitro approaches 1 , P.P. , Fiore Di nb/nbl 2 , erythropoietic hierarchy.erythropoietic Meis1-depleted embryos. Combined, these results place Pbx and Meis1 upstream of 1 L.M. Reaume The HoxcofactorsPbxandMeis1actupstreamofgata1toregulateprimitive erythropoiesis Hematopoiesis andimmunology inhibit myelopoiesis. of upstream Meis1, and Pbx with association in acts Hox whereby model a propose therefore We embryos. Meis1-depleted not Furthermore, cells. myeloid maintain of loss a with Concomitant fate. of expression the in reduction severe a exhibit and cells, blood circulating visible produce to fail Meis1 and Pbx lacking Embryos Meis1. and cofactors Pbx Hox the ablated have we hematopoiesis, primitive zebrafish in function Hox of loss cofactors Hox as play that proteins role TALE-class significant of loss the from results phenotype identical Meis An 1. and rhombomere of identity the on take Homeobox) 2-6 rhombomeres in which in phenotype hindbrain, zebrafish anteriorizing the Leukemia an generates Pbx (Pre-B-Cell of Removal Site). Pbx Integration Ecotropic (Myeloid factors transcription homeodomain class with other DNA-binding cofactors. These include the transcriptional gene networks. The specificity of Hox proteins isposterior achieved erythroid of through of their aspects interaction certain regulating in factors lineage specification.However, theoverlappingexpressionpatternsandfunctionalredundancy transcription these for role a demonstrating Vertebrate embryos bearing deletions in individual Halifax, University, Dalhousie Canada Centre, Health IWK Microbiology/Immunology, and Pediatrics Department of Biological Sciences, University of Alberta, Edmonton, Canada Edmonton, Alberta, of University Sciences, Biological of Department hemangioblast gene expression, and possess increased numbers of numbers increased possess and expression, gene hemangioblast scl genes have prevented us from defining their role in regulating hematopoietic regulating in role their defining from us prevented have genes hox 1 , T. Erickson overexpression rescues hoxb7a-overexpression 1 , A.M. Forrester vrxrsin s be o drive to able is overexpression gata1 gata1, Px n Mi1dpee ebys ntae bt al to fail but initiate, embryos Meis1-depleted and Pbx , gata1 to specify the primitive erythropoietic cell lineage and lineage cell erythropoietic primitive the specify to 2 , J.N.Berman ee rdcs in products gene Hox1 hox genes display a partial loss of blood cells, , the earliest marker of erythroid cell erythroid of marker earliest the , gata1 Three 2 expression in Pbx-depleted, but Pbx-depleted, in expression gata1 , A.J. Waskiewicz Amino acid . In order to model global model to order In vivo. in 270 xrsin n b and Pbx in expression scl idctn the indicating Xenopus, Loop 1 Extension (TALE)- 2 Department of Department pu.1-positive gata1 in the 341

Posters Posters 342 the identificationofnoveltherapeuticentities. for compounds lead useful be may compounds these of Some properties. immunomodulatory Conclusions: mammalian in inflammation of resolution models ofneutrophilicpulmonaryinflammation. of enhancement for assessed be will and function These compounds are under further investigation for their ability to modulate human neutrophil one-way ANOVA withBonnferronipost-testcorrection,n=49,24and9respectively). and for one compound it was 12.33+/-1.85 (mean+/- sem, p<0.05 for both treatments vs control, of injury at 24 hours post-injury was 30.24+/-1.47. In pyocyanin treated fish it was site the at 19.04+/-1.93, present neutrophils of number the fish control in example, For pyocyanin. apoptosis, inflammation to levels below those seen with our positive control, a potent inducer of neutrophil tested and shown to exhibit dose-dependent reproducible effects. Active compounds suppressed neutrophilic have been have number to a these, shownOf agents. anti-inflammatory were known several include 12 These effects. which of tested, were compounds 960 (MSdiscovery), utility of this approach in the identifcation of new immunotherapeutics. From the Spectrum collection the inflammation demonstrating principle”, of “proof as accelerate serve that compounds lead several which identified compounds have and screens, such of practicality the demonstrated identify have experiments Preliminary Results: to Thispermits time. libraries over resolution. compound resolves of spontaneously screening which quantifiable and response reproducible a inflammatory to leads neutrophilic larvae anaesthetised of tailfin the to injury physical quantitated rapidly be can resolution inflammation which in lineage, neutrophil the in GFP toenhanced expressing zebrafish transgenic in model tractable a established Wehave Methods: leading thisapoptosis delay signals inflammation. survival diseases inflammatory during but apoptosis, undergone having macrophages by removed usually are Neutrophils diseases. such treat to ways new identify to need unmet major a is There therapies. current to poorly respond Rationale: Biomedical and Developmental for Centre Genetics, University ofSheffield, MRC Western Bank, Sheffield, and UK Medicine Respiratory of unit Academic C. A. Loynes, A. L.Robertson,M.K.B.Whyte,P. W. Ingham,S. A. Renshaw Using transgenic zebrafish toscreenforsmallmoleculeinducersofinflammationresolution 271 Diseases of neutrophilic inflammation are common, affect many organ systems, and systems, organ many affect common, are inflammation neutrophilic of Diseases These data show the ability of this model to identify novel therapeutics with dramatic Hematopoiesis andimmunology . Sterile vivo . in subset markersisinvestigatedinthedevelopingandadultzebrafish. vertebrates over hundreds of millions of years of evolution. Using quantitative PCR (Q-PCR), PCR quantitative Using the Whole evolution. among of years order of gene millions of maintained hundreds has over striking vertebrates selection a that exhibit suggesting genes order, these gene vertebrates, of other conservation with compared when level, sequence the at STAT6, Th-POK, CD8, T-bet, CD4, identified and have FoxP3 we in zebrafish. approach, Despite synteny high a levels of and divergence genome withzebrafish somethe ofUsing these genes of lowervertebrates. specific and time specific mannertissue would begina to revealin important detailsdistribution of thetheir immune system at looking and T-cell play subsets of development that during factors role critical transcription a these Using mammals. of those parallel fish in subsets T-cell of properties functional and development that conclude firmly cannot we T-bet)means & FoxP3 absence of other cell markers, which include T-cell cells specific and Treg the transcriptionteleosts, Th17 in characterised been recently have factorsCD8 and Th2, CD4 mammals. Although in as exist, (Th-POK, Th1, STAT6, as such subsets cell whether and fish in development cell (Th) understand to model vertebrate useful normal white blood cell development. Despite this, a however, very little is known as about T-helper recognized now and immunity is ) rerio (Danio zebrafish The Aberdeen, UK Sciences,Biological of Centre(SFIRC),ResearchUniversityof Fish Immunology School Scottish S. Mitra, A. Alnabulsi, C.SecombesandS.Bird rerio) Discovery and expression analysis of important markers of T-cell subsets in the zebrafish (Danio Hematopoiesis andimmunology In Situ In Hybridisation (WISH) and a FoxP3 polyclonal antibody, the expression of T-cellof expression antibody,the polyclonal FoxP3 a and (WISH) Hybridisation considerable progress has been made in understanding the molecular basis of basis molecular the understanding in made been has progress considerable 272 343

Posters Posters 344 as such genes of up-regulation through vessel, the of induction wall ventral the to in cells in programme leads HSC the of then This aorta. dorsal the in activity notch induces turn in which somites, neighbouring in Vegfsignalling induces notochord the from signalling Hedgehog that is thought it definitive zebrafish, the of In embryos. formation zebrafish the and mouse in both in role (HSCs) cells important stem an haematopoietic play to shown been has pathway Notch The embryonic in Institute ofGenetics, targets NottinghamUniversity, UK downstream and receptorsK. A. McMaahon,J.Rowlinson,M.Gering ligands, notch of haematopoiesis andangiogenesis roles the Elucidating genes inregulatingangiogenesis. search for other putative Notch targets in the dorsal aorta has revealed a role for two Notch targettargetnotch putative the pathway.the of regulator upstream an fact in is Gridlock a addition, In that suggesting data present Wealso programme. HSC the of induction for responsible ligands morpholino knockdown and over-expression studies, we Using have haematopoiesis. identifieddefinitive theinitiating notchin receptorsNotch and of role the of dissection detailed a present Weelucidated. be to yet have process the regulates pathway this which by mechanisms exact the HSCs, zebrafish of formation the in identified been has Notch for need general a Although embryos leadstoectopicexpressionofrunx1intheaorticroofandvein. zebrafish in domain intracellular notch of Conversely,overexpression aorta. dorsal the in cells notch signalling in the E3 ubiquitin ligase mutant 273 and runx1 . In support of this, previous work has shown that down regulation of regulation down that shown has work previous this, of support In c-myb. mindbomb, leads to a loss of Hematopoiesis andimmunology runx1 expressing Cellular Medicine, CMVB, Katholieke Universiteit Leuven, Belgium;Leuven, Universiteit Katholieke CMVB,Medicine, Cellular 1 ope rcpos GCs. ncie eeormrc -rtis r cmoe of of composed are differentiation G-proteins heterotrimeric the Inactive (GPCRs). receptors involves coupled hematopoietic that by controlled process are growth lymphopoiesis factors, developmental and including Myelopoiesis cytokines, cells. life-long and stem chemokines that hematopoietic a bind to seven-transmembrane is G-protein- Hematopoiesis Katholieke CMVB, Medicine, Cellular and Universiteit Leuven, Molecular Belgium of Department and Child section Child, S. Louwette The roleofRGS18inhematopoiesisandmegakaryopoiesis Hematopoiesis andimmunology and thrombocyteformation. In conclusion, this study showed for the first time a role for an RGS protein in megakaryopoiesis The secondaimofthestudyistounravelpathwaybywhich RGS18isworking. ofhematopoiesis/ phase in which out find megakaryopoiesis RGS18isinvolved. to needed are hematopoietic experiments early to additional related not but were defects KD the of effects the that showed results situ In The blot. formation. somite and vessel- and Western analysis cytometry flow blood by confirmed was thrombocytes GFP-labeled in in decrease defects secondary and thrombocytes total of in decrease number dependent dose a caused MOs Both immunoblot. by confirmed was KDs in RGS 18, were injected of in one region ATGcell the stage in eggs of knock sequences CD41 different (MO) two morpholino against developed the Twotechnology.MOs, down used andisalso we megakaryopoiesis, in RGS18 of role the in understanding step initial an inmegakaryocytes, As receptors. Gq-coupled from signals attenuate and Gαi, butits of activity GTPase abundant cells, enhance vitro, ishighly in Gαq lineage and Gαi both andmyeloerythroid bind can RGS18 megakaryopoïesesknown. not in is role progenitors RGS18 hematopoietic amodel. in specifically detected as zebrafish using Our lab mainly focuses on studying the role of RGS 18 in hematopoiesis and megakaryopoiesis, lymphoid cells. or myeloid mature of function the in implicated been have for proteins RGS proteins Several subunits. GTPase-activating Gα as signaling act can G-protein that family of protein Regulators discovered recently subunit. a are Gα (RGSs) the of activity GTPase intrinsic the increasing by pathways G-protein heterotrimeric regulate can of proteins reformation GTPase-activating G-αβγ. to inactive GDP,leading to GTP bound hydrolyses Gα-subunit the when terminated is active of formation the in GTP,bind to GDPand release to Gα stimulate resulting GPCRs binding, ligand Upon subunits. redeecnrm ahlee nvriet evn Belgium; Leuven, Universiteit Katholieke Proefdierencentrum 1,2 , C.Wittewrongel wih euae ontem fetr. Signaling effectors. downstream regulate which G-βγ, and α-GTP 2 , C.

Vangeet 2,3 , J. Arnout + -GFP zebrafish. Downregulation of RGS18 1 , K.Freson 2 eatet f oeua and Molecular of Department 274 3 2 Department of Woman & Woman of Department α, and β 345 γ

Posters Posters u t ter at ifso ad estl booia atvte, ecie xgn pce (ROS) species oxygen reactive activities, biological versatile and including hydrogen peroxide diffusion (H fast their to Due response areunknown. tissue immediate this direct that signals spatial the of characteristics biophysical and molecular to leukocytes The wounding. of minutes within of micrometers of hundreds of distances across injury of site recruitment the barrier include restore steps initial to animals, executed In homeostasis. rapidly tissue be and integrity can that program morphogenetic are dormant cells) a around represent membranes plasma tissues, around required for internal homeostasis and protection from pathogens. Wound detection and healing epithelia (e.g. structures barrier Eggenstein-Leopoldshafen, Intact GmbH, Karlsruhe Forschungszentrum Technology,Germany of Institute of leukocytes to the wound. This is the first observation of a tissue-scale H tissue-scale a of observation first the is This wound. the to leukocytes of that this gradient is created by dual oxidase (DUOX), and that it is required for rapid recruitment as a decreasing concentration gradient. Using pharmacological and genetic inhibition, we show after wounding and peaking at ~20 min that extended ~ 100-200 μm into the tail fin epithelium We measured H knockdown, morpholino using perturbed tissue thetransgenicexpressionandpharmacology. of dynamics molecular the and imaged, and regenerative responses to wounds. Rapid leukocyte recruitment to the wound can be easily The zebrafish larval tail fin has become a popular vertebrate model system to study inflammatory together with leukocyte motility, in an intact vertebrate tissue subjected to mechanical wounding. nooy Dn-abr acr nttt, avr Mdcl col Bso, USA; Boston, School, Medical Harvard Institute, Cancer Dana-Farber Oncology, selective for H for selective 346 C. Grabher A tissue-scalegradient ofhydrogen peroxidemediatesrapid wound detectioninzebrafish 1 first evidencethatH excited at 420 nm (YFP nm 420 at excited This reporter revealed a sustained rise in H HyPer bymRNA injectionintozebrafishembryostoinduceglobalcytoplasmic expression. environment, allowing dynamic monitoring of intracellular H conformational change that increases emission excited at a induces 500 part OxyR YFP.the permuted nm circularlyof a oxidation to Cysteine (YFP fused OxyR factor To investigate possible paracrine signalling by H by signalling paracrine possible Toinvestigate Departement Departement of Systems Biology, Harvard Medical School, Boston, USA; 275 2,3 , P. Niethammer 2 O 2 O 2 over other ROS. HyPer consists of the bacterial H bacterial the of consists HyPer ROS. other over 2 by expressing HyPer, a genetically encoded ratiometric sensor that is highly 2 O 2 signalstoleukocytesintissues,additionitsknownantiseptic role. 420 ). This change is rapidly reversible within the reducing cytoplasmic reducing the within reversible rapidly is change This ). 1 , A. T. Look 2 O 2 ) are interesting candidates for wound-to-leukocyte signaling. 2 2 O , T. J.Mitchison 2 concentration at the wound margin, starting ~3 min 2 O Hematopoiesis andimmunology 2 , we imaged its spatiotemporal dynamics, spatiotemporal its imaged we , 1 2 O 2 concentration. We introduced 500 2 O ) and decreases emission 2 Department Department of Pediatric 2 -sensitive transcription -sensitive 2 O 2 pattern, and the and pattern, 3 Karlsruhe A.T. Look Department of Molecular Genetics, University of Toronto,Canada; of University Genetics, Molecular of Department Zebrafish miR-126 and miR-150 coordinately determine hematopoietic cell fatethrough cell Grabher C. hematopoietic determine coordinately c-Myb miR-150 and miR-126 Zebrafish Hematopoiesis andimmunology otn USA; Boston, during hematopoiesis. but also are essential determinants in the control of variable potential functions of a single gene support the notion that microRNAs not only act to provide precision to developmental programs between decision fate erythroid cell the and regulate precisely megakaryocytic to individually lineages sufficient not during but required sequentially definitive hematopoiesis. Our observations is adjusted by the combined activity of the miRNAs miR-126 and miR-150. Both microRNAs are lineages cell erythrocyte versus thrombocyte of specification that evidence provide Wefurther of c-myb alleviatestheseeffects. promotes erythropoiesis at the expense of and thrombopoiesis levels protein c-Myb increased in results miR-126 of Knockdown region. coding upstream specifically binds to a predicted target site within the miR-126 zebrafish. that the show in We hematopoiesis definitive during 126/c-mybinteraction Here, we demonstrate the direct regulation of c-Myb by miR-126 and the importance of the must bepreciseandtightlymaintained. activity c-Myb of levels and timing the controlling mechanisms the roles, developmental its of diversity spatio-temporal the Given lineages. megakaryocyte and erythroid myeloid, lymphoid, and progenitor cells to fully differentiated cell types, and can affect the definitive specification of proto- oncogene of The development. levels cell blood cellular during decisions the fate cell modulate controlling factors appropriately transcription to required are mechanisms regulatory Precise Oncology, ofPediatrics, Department Children’s Hospital, MedicalSchool, Harvard Boston, USA­ 1 eatet f eiti Oclg, aaFre Cne Isiue Hrad eia School, Medical Harvard Institute, Cancer Dana-Farber Oncology, Pediatric of Department 1, 3 functions at multiple stages during hematopoiesis, from hematopoietic stem- hematopoietic from hematopoiesis, during stages multiple at functions c-myb 1 EM Payne E.M. , 2 iiin f el n Mlclr ilg, nvriy elh ewr, and Network, Health University Biology, Molecular and Cell of Division 1 AB Johnston A.B. , 1 N Bolli N. , c-myb 3’UTR to attenuate translation of its 1 E Lechman E. , in vivo, while concomitant knockdown 276 3 2 Division of Hematology/ of Division JE Dick J.E. , 2 JP Kanki J.P. , miR- 347 1 ,

Posters Posters 348 the at report progress a present will and further lines meeting. the characterizing currently are We of theDA. reporter gene expression in primitive erythrocytes, in myeloid cells as well as in the ventral wall interest. One transgenic line has expression in primitive erythrocytes while a second line shows label and non- fluorescently endothelial tissues, we have also found two lines that show to GFP expression patterns of b) haematopoietic non- various in expression gene tissue-specific with and lines transgenic a of number identifying Besides haematopoiesis development. of stages and different at cells haematopoietic vasculogenesis and endothelial in involved genes novel identify to a) aims the with screen trap gene/enhancer based transposon a final conducted Wehave their seeding before tissue haematopoietic caudal destination, thekidney, the thesiteofadulthaematopoiesisinfish. in settle vein, the via circulation association close with the in ventral wall form of the dorsal HSCs aorta (DA). vertebrates, The cells other subsequently enter in the blood As embryo. the of trunk the in mass cell intermediate the in occur haematopoiesis of wave definitive the and erythrocytes primitive the zebrafish, In that differentiateintomaturecellsofallbloodlineages. progenitors to rise give to and self-renew to able are HSCs adulthood. throughout system blood and respectively. theanterior The definitive wave from generates haematopoietic arises stem cells wave (HSCs), first whichposterior maintain The lateral the plate waves. mesoderm two which give in rise occurs to haematopoiesis primitive Vertebratemyeloid and to primitive erythrocytes Centre, Medical Queen’s Nottingham, of University United Kingdom Biology, of School Genetics, of Institute R. Thambyrajah andM.Gering in haematopoieticandendothelialCells Gene/ Enhancer trap based screen identifies novel transgenic lines with reporter gene expression 277 Hematopoiesis andimmunology hybridization with gene markers of hematopoietic cells. At st32, the cells. st32, hematopoietic At of markers gene with hybridization situ in of onset the for gene marker a as definitive hematopoiesis. used widely To analyze is the phenotypes and of hematopoiesis definitive for essential is decreased expression of erythrocyte marker revealing alinkbetweenprimitiveanddefinitivehematopoiesis. arises. This result suggests that complete loss of the definitive hematopoiesis is malignant. Surprisingly, in the early stages of st22, we observed a that indicate which myeloid other in also but erythrocytes in only not defects have mutants bef marker,neutrophil of expression the the that revealed cloning Positional-candidate blood. the in mutation nonsense a have mutants peripheral the in erythrocytes of all generate that number reduced apparently has that mutant medaka a cell as isolated originally was (bef), fuji Beni stem hematopoietic of emergence hematopoietic lineage. the with accompanied the stages in place later takes that hematopoiesis definitive and macrophages, generate and that erythrocytes embryos primitive early the in phenomenon transient a is which hematopoiesis primitive Vertebrate hematopoiesis is characterized by two evolutionary conserved phase of development; Radiation Protection, National InstituteofRadiologicalSciences A. Moriyama progenitor differentiation;aninsightintohematopoieticontogenesisinmedaka Characterization of medaka Hematopoiesis andimmunology 1 eatet f ilgcl nomto, oy Isiue f Technology of Institute Tokyo Information, Biological of Department 1 , K.Maruyama l-plastin expression in the anterior region from where the primitive macrophage 2 C-MYB mutant , A. Kudo c-myb may also function in the primitive hematopoiesis, potentially was completely lost. These results suggest that the that suggest results These lost. completely was mpo1 1 gene. Previous studies have shown that shown have studies Previous c-mybgene. α-globin, and myelomonocyte marker BENI FUJI, bef mutants, we performed whole-mount which displays defective hematopoietic 278 2 eerh etr for Center Research bef mutants showed mutants l-plastin, and c-myb 349 bef

Posters Posters ees Uiest, eh, api Tia 14 R.O.C.; 114, Taiwan Taipei, Neihu, University, Defense 1 350 S-P.L. Hwang β-lapachone treatmentcausesapoptosisofredbloodcellsinzebrafish embryos ROS mediatedapoptoticpathway. produced in of doze lethal sub detectable possessing tissues these on effects toxicity no were There stages. developmental different from embryos of bulb intestinal and line, lateral posterior of neuromasts otoliths, arches, pharyngeal bulbs, olfactory epidermis, the in detected mRNAis development of T lymphocytes were also affected in in phenotype deficiency cell blood red rescue could BAPTA-AM,chelator, calcium or dicumarol, inhibitor, in red blood cells of hpf of (2μm) doze lethal pericardial edema were detected in 48-52 hours post fertilization (hpf) embryos treated Linear with sub unknown. is development arrangement of embryonic ventricle and atrium, no circulation or circulation on with few red blood cells, and effect its while adenocarcinoma, pancreatic of treatment the for trials clinical II phase in currently is agent, antitumor novel a β-lapachone, Biology (formerlyInstituteofZoology), Academia Sinica, Nankang, Taipei, Taiwan 115, R.O.C. 2, R.O.C.; 221, cdma iia Nnag api awn 1, R.O.C.;Taipei, 115,Taiwan Nankang, Sinica, Academia Institute of Bioscience and Biotechnology,and TaiwanBioscience National University,of Ocean Institute Keelung, Taiwan 279 β-lapachone-treated embryos. There were abundant ROS production and DNA fragmentation β-lapachone-treated zebrafish embryos is also mediated through NQO1-dependent, 1,4 2 lpcoetetd mro. el ubr f etohl rnlcts n the and granulocytes neutrophil of number Cell embryos. β-lapachone-treated rdae nttt o Lf Sine, ainl ees Mdcl Center, National Medical Defense National Sciences, Life of Institute Graduate , Y-T. Wu -lapachone. These results indicate that red blood cell deficiency phenotype deficiency cell blood red that indicate results These β-lapachone. β-lapachone-treated embryos. Co-treatment -lapachone. Severe yolk and pericardial edema was then detected in 96 in detected then was edema pericardial and yolk Severe β-lapachone. 1 , C.Y. Lin 1 , M-Y. Tsai 2 , Y-H. Chen β-lapachone-treated embryos. Visible Hematopoiesis andimmunology 4 4 , Y-F. Lu Institute of Cellular and Organismic Organismic and Cellular of Institute 3 nttt o Booia Chemistry, Biological of Institute mRNANQO1 with treated when β-lapachone with either NQO1 4 , C-J.Huang 3

NQO1 are crucialfornevi/melanomatransformationandmelanoma progression. pathways signaling which and interact, spectrum UVR and pigment how melanoma, initiating for responsible are repair) DNA and photoproducts (e.g. events molecular which addressing of capable be will we system, model zebrafish our With exposure. UVR after melanoma develop tumor- pigmentation, for to altered likely are lines mutant these Weof oncogenes. some or that genes reasoned are suppressor have lines These UVR lab. the after in susceptibility available lines melanoma mutant and different in ability repair damage DNA the investigating are We are alsoimportantfortheresponsetoUVRinzebrafish. to similar experiments in mouse knockouts. This is further evidence that cancer genes in humans had only 31% of (6-4) PDs repaired. (AB The p53 fish zebrafish showed reduced NER of wildtype (6-4)PDs, comparable (hpr), mutant p53 whereas NER, DNAby epidermal radiation their from removed PDs (6-4) of post 63% had strain) hours 24 At irradiation. UVB after skin zebrafish adult DNAof the in induced PDs) ((6-4) dimers pyrimidine-pyrimidone 6-4 and (CPDs) dimers pyrimidine cyclobutane photoproducts, major two the measured we (RIA), radioimmunoassay irradiation, indicating that genetic differences can affect the response to UVR in zebrafish. Using to the one used in the Using as welltheabilitytousesmallmoleculesmodulateresponseUVR. elucidate the genetic changes that occur after UVR treatment and UVR-induced melanoma. Advantages of a zebrafish UV-melanomaduring model include the ability to melanoma development, that of a from translated are melanoma UVR-induced of technology and knowledge our of Part model. melanoma UVR-induced zebrafish a generate to aim the with backgrounds, genetic different in of melanoma. We are studying the pathological effects of Ultraviolet irradiation (UVR) UVR-treatment is the on most important melanocytes environmental risk factor and for the nevi development Anderson CancerCenter, Smithville, Texas, USA 1 Z. Zeng Development ofzebrafish modelsofUVinducedmelanoma Diseases models of Edinburgh, United Kingdom; United Edinburgh, of and p53 mutant zebrafish is near 480J/M near wildtype is zebrafish mutant p53 and 6-day-old for genetic irradiation UVB different of with LD50 zebrafish the showed of results ability Preliminary backgrounds. (NER) repair excision nuclear and dosage UVR the Institute for Genetics and Molecular Medicine, MRC Human Genetics Unit and The University and The Unit Medicine,Genetics Molecular Human and MRC Genetics for Institute Xiphophorus as a foundation for our work, we have built a UV irradiation chamber similar 1 , E.Patton Xiphophorus hybrid ( 1 Xiphophorus D. L.Mitchell X. maculates Jp163B × 2 Department of Carcinogenesis, The University of Texas M.D.Texas of University The Carcinogenesis, of Department melanoma models. Experiments were carried out to determine 2

2 , but p53 mutant fish had a delayed response to UVB to response delayed a had fish mutant p53 but , X.helleri ), a well established fish model for 280 351

Posters Posters 352 ERGs function. Our data suggest that zebrafish are a useful animal model for research into blood-barrier diabetic retinopathy. retinal the of confirm defectsinconephotoreceptorfunctiontheglucosetreatedfish. breakdown a a displaying indicating thicker, are is abnormal, basement the membranes and open, more significantly are histology junctions interendothelial the retinal animals, treated fish, glucose-treated of pronounced degeneration of cone photoreceptors. In regards to the retinal capillaries of glucose ~60% In retinopathy. diabetic stage early of hallmarks vascular and neurodegenerative display fish Glucose-treated assess treatment, to conducted electrophysiological functioninthetreatedretinas. of were days (ERG) electroretinograms 30 Ex-vivo microscopy. After electron control. transmitted osmotic an as onalternative used retinal morphology and retinal vascular network were analyzed by immuno-histochemistry and was 2%glucose treatment with mannitol treating 2% by days. zebrafish adult in induced was Hyperglycaemia vasculature. is nourished by an intricate vascular network which shares many features with the human retinal zebrafish adult in retina inner The patients. diabetic of retinas the in observed microangiopathy vascular and damage neuronal the recapitulating changes functional and/or morphological of induction the through retinopathy diabetic of model zebrafish a develop to was objective Our animal modelsofNPDRareessential. thus, patients for asymptomatic mostly are retinopathy diabetic of stages Early damage. retinal non-proliferative phase of the disease (NPDR) leading may reveal interventions the that more effectively and limit progression of proliferative diabetic diabetes retinopathy (PDR) but a better comprehension of the initial, 2 type of complication palliate can treatments Some countries. developed in people working-age in blindness of cause common most the is retinopathy Diabetic Conway Institute, UCD, Dublin, Ireland Y. Alvarez, K.Chen, A. Reynolds,N.Waghorne, J.O’ConnorandB.Kennedy Recapitulating EarlyStagesofDiabeticRetinopathy inHyperglycaemicZebrafish 281 Diseases models to change in ageing cells, therefore, aberrant splicing of particular genes may play a role in role a play may genes particular of sporadic formsof AD andotherneurodegenerativediseases. splicing aberrant therefore, cells, ageing in change to Ccng2 with interfere Ccng1 of forms Potentially,truncated Ccng1. the truncating as development embryo on effects similar in (CCNG2), normal of level the decrease not does it of Ccng1 appears to cause developmental defects in the brain, notochord and somites, however, aberrant splicing of defects and also cannot rescue the hydrocephalus or pigmentation phenotypes of embryos with developmental any to lead not does expression Ccng1 Decreased somites. and tectum eye, the zebrafish of synteny analysis confirmed the orthology of zebrafish of orthology the confirmed analysis expression. synteny increased showed and immunoreactivity function in the cytoplasm of neurons in known human AD brains. Phylogenetic and have conserved these of Cyclin Five treatments. both to point mutations and in splicing the APP.Aberrant human of presenilin domain genes, intramembrane the of cleavage gamma-secretase amyloid of result a precursor as protein brain, (APP) cleavage the by in secretase enzymes. peptides The presenilin beta proteins amyloid are central accumulating to by the caused is AD that Alzheimer’s disease (AD) is the most prevalent form of dementia. There is considerable evidence Molecular andBiomedicalScience, The University of Adelaide, Australia M. Newman,S.Nornes,B.Tucker andM.Lardelli Using zebrafish embryos toinvestigate genesimplicatedin Alzheimer’s diseasepathology Diseases models changes to illuminate the molecular aetiology of these phenotypic effects. Of the 100 genes 100 the Of effects. phenotypic after dysregulation greatest showed that these of aetiology molecular the illuminate to changes 8 exon of splicing with interference in low-level after observed also are effects Similar defects. developmental other and pigmentation decreased phenotype, hydrocephalus dramatic a cause zebrafish embryos produces potent dominant negative effects that increased embryos. We have demonstrated that low-level aberrant splicing the of have exon understand 8 to genes in aim these We disease. role Alzheimer’s in occur that pathways complex the of some human of orthologues AD, through aberrant APP cleavage resulting in irregular amyloid beta formation. Zebrafish have psen2 a o priua itrs a te ua CN1 rti sos increased shows protein CCNG1 human the as interest particular of was (ccng1) G1 transcripts. A microarray analysis was performed to analyse global gene expression gene global analyse to performed was analysis microarray A transcripts. ). Decreasing the expression of Ccng2 results Ccng2 of expression the Decreasing (ccng2). zebrafiish in expressed also is in developing embryos at 24 hours post fertilization (hpf) was observed in observed was (hpf) fertilization post hours 24 at embryos developing in ccng1 psen1 exon 8. Analysis of zebrafish and 1 PSEN nA ptooy y netgtn ter oeua booy n zebrafish in biology molecular their investigating by pathology AD in function in a dominant negative manner. Splicing is observed is Splicing manner. negative dominant a in function and 2 or psen1 PSEN1 and transcript. The transcript. ccng1 , therefore they are a useful tool for investigating for tool useful a are they therefore APP, manipulation, 12 genes were common were genes 12 manipulation, psen2 PSEN2, have been linked to familial forms of with human with ccng1 ccng1 function indicates that truncation paralogue, CCNG1 282 . Expression CCNG1. psen1 transcription, psen1 transcripts in Cyclin G2, Cyclin 353

Posters Posters brain inrelationtoNotchand APP signalingpathways. adult in and embryogenesis during compartment cell stem neural the FAD-PSEN1on of effects human FAD-PSEN1 under the zebrafish FAD-PSEN1transgenic tagged novel mCherry-fluorescently generated expressing have lines we and PSEN1L286V, neutral towards Notch Psen1-null in mRNA FAD-PSEN1 human of embryos, we aim to dissect the misexpressioneffects of the PSEN1M146V, By decreasing Notch signaling brain. adult the in of-function a As loss- Psen1 FAD-PSEN1to life. of respect effects with study to opportunity with unique a have we result, compatible PSEN1 of knockout gene mutant targeted with Psen1-null model vertebrate zebrafish first The the is impaired. is signaling Delta/Notch addition, In cerebellum. and proliferation cell decreased show 354 Psen1. functional lacking mutant These Psen1-null mutants are viable zebrafish and fertile, in contrast to characterized recently our utilize we end, this To model a as zebrafish system. using brain adult and embryonic the in mutations FAD-PSEN1 different such of expression of effects the reveal to Weaim neutral. are others whereas signaling Notch neural stem cell niche and neurogenesis in adult. Some FAD-PSEN1 mutations lead to impaired a protein essential for correct neuronal differentiation during embryogenesis, maintenance of the PSEN1- by mediated formed proteolytic is cleavage of A β the accumulation. amyloid (Aβ) precursor protein amyloid-β (APP). by PSEN1 also characterized cleaves neuropathologically Notch, Autosomal dominant mutations in presenilin 1 (PSEN1) lead to familial Alzheimer’s disease (FAD), Utrecht, Utrecht, The Netherlands Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences & University P.Medical Centre van Tijn, J.T. Paridaen,C.vanRooijenandD.Zivkovic Zebrafish modelsforfamilial Alzheimer’s disease 283 neurogenesis, which is most pronounced in the in pronounced most is which novoneurogenesis, de psen1 promoter region. In these lines we are investigating , mutations in during vitro embryogenesis. In addition, PSEN1 -/-

mice. Psen1-null mutants Diseases models in , vitro a gooddiseasemodelforPD. zLRRK2 is a true ortholog of hLRRK2 and that the deletion of WD40 domain of zLRRK2 provides Takenneurodegeneration. not but defects, locomotion together,that indicate clearly results our over-expressing zLRRK2 or hLRRK2. Interestingly, administration of L-dopa could also rescue the midbrain in by rescued be could defects locomotion and neurodegenerative These neurons defects. locomotion and dopaminergic of loss including phenotypes, Parkinsonism-like caused it rather defects; developmental embryonic severe induce not did splicing targeting morpholinos growth retardation and loss of neurons. In contrast, deletion of the WD40 domain of zLRRK2 by expression by morpholinos caused are embryonic lethality and functions severe developmental biological defects such its as expression its and characterized investigated but its (zLRRK2), biological LRRK2 functions in human (PD), zebrafish.of The blockagehomolog disease ofthe zLRRK2cloned protein Parkinson’swe Here, unknown. in largely role important an plays LRRK2 Singapore S. Jesuthasan . Sheng D. Deletion ofthe WD40 domainofLRRK2inzebrafish provides amodelfor Parkinson’s disease Diseases models 1 Duke-NUS Graduate Medical School.Medical Singapore;Graduate Duke-NUS eoe nttt o Snaoe ASA, Singapore; A*STAR, Singapore, of Institute Genome ua Genetics, Human 1 D Qu D. , 5 , M.Sinnakaruppan 1 2 SS Ng S.S. , eeomna Booy and Biology Developmental 1 YMA Lim Y.M.A. , 2 , J.J.Liu 1 2 WHC Lee W.H.C. , 5 Neuroscience Research Partnership,Research Neuroscience A*STAR, 3 opttoa ad ahmtcl Biology, Mathematical and Computational 4 ainl ersine nttt and Institute Neuroscience National 3 SW Kin S.W. , 284 3 EK Tan E.K. , 4 T Lufkin T. , 355 2 ,

Posters Posters some oftheaspectspontocerebellarhypoplasia. phenocopy may morphants that suggest data These development. brain affecting prominently and splice site morpholino oligonucleotides and observed neurodevelopmental phenotypes, hypoplasia. As most a first step we performed knock down of TSEN54 or TSEN2, employing both ATG mechanisms underlying PCH we are currently establishing zebrafish models for pontocerebellar highlyThe molecular pathways behind is pontocerebellar hypoplasia remain TSEN54 unclear. In that order to study revealedoftRNA-Tyrosinethe fromfibroblastsof3patientsdidnotshowunsplicedproducts. probes LNA/2OME analysis blot Northern nuclei. olivary and dentate using cerebellar pons, the of neurons in TSEN54expressed for hybridization situ In identified missenseandnonsensemutationsinTSEN54,TSEN34TSEN2subunits. EuropeanPCH2 also plays a role of in pre-mRNA 3’end formation. majority In PCH patients without the inthe A307S mutation, we and tRNAs containing intron of splicing the for responsible is complex TSEN The TSEN15). and (A307S) inTSEN54 patients. TSEN54 is one mutation of the four subunits of the tRNAa common splicing endonuclease (TSEN34, TSEN2 identified We reach adulthood. and variable neocortical atrophy. The disease microcephaly is progressive and usually pons, patients die before they ventral the of hypoplasia cerebellum, the of hypoplasia or atrophy to due disorders with prenatal onset (PCH1-6). Children suffer from Pontocerebellar hypoplasia (PCH) represents a group of neurodegenerative autosomal recessive severe mental and motor impairments Academy of andSciences &University MedicalCentreArts Utrecht, The Netherlands A.J. Grierson Germany; and Biomedical Sciences, University of Sheffield, UK; Sheffield, of University Sciences, Biomedical and 1 356 Kasher P. tRNA SPLICINGENDONUCLEASEMUTATIONS CAUSE PONTOCEREBELLAR HYPOPLASIA Netherlands; University of Amsterdam, The Netherlands; The Amsterdam,of University etr Asedm Te Netherlands; The Amsterdam,Center, eatet f ergntc, cdmc eia Cne, nvriy f mtra, The Amsterdam, of University Center, Medical Academic Neurogenetics, of Department 285 1 . Namavar Y. , 3 iiin f adarc erlg, ma hlrns optl Aaei Medical Academic Hospital/ Children’s Emma Neurology, Paediatric of Division 5 , P. vanTijn 2 oon Cne o Gnmc ad nttt o Gntc, nvriy f Cologne, of University Genetics, of Institute and Genomics of Center Cologne 1 BS Budde B.S. , 6 , F. vanRuissen 2 PG Barth P.G. , 4 eatet f ahlg, cdmc eia Center, Medical Academic Pathology, of Department 1 , M.Weterman 5 Academic Unit of Neurology, School of Medicine of Neurology,School of Unit Academic 3 B Te Poll-The Tien B. , 6 urct nttt, oa Netherlands Royal Institute, Hubrecht 1 , D.Zivkovic 3 K Fluiter K. , 6 , P. Nürnberg Diseases models 1 , E. Aronica E. , 2 , F. Baas 1 4

, 1 T. van Boxtel Dithiocarbamates areteratogenic tozebrafish throughinhibitionoflysyloxidases Diseases models of DTC. caused by direct inhibition of LOX activity. These findings have implications for risk assessment of knockdown Taken partial together, these results that exposure. indicate that teratogenic effects caused by DTC DTC are, at least in part, to demonstrate embryo developing the and sensitizes loxl5b and vitro loxl1 lox, in oxidases, lysyl three LOX inhibit directly DTC that show we bydirect Additionally,(LOX). oxidases lysyl termed enzymes observed containing copper of effects group a of teratogenic inhibition resemble highly that zebrafish in malformations cause chelators but this does not fully explain the observed teratogenic effects. We show here that DTC distorted notochord development and shortened anterior to posterior axis. DTC teratogenic to developing vertebrates. In zebrafish these teratogenic effectsare are characterized by known copper they that established well is it lives half short their to due safe relatively be to thought are DTC pesticides. As such humans and wildlife are undoubtedly exposed to these chemicals. Although as agriculture in used extensively are which compounds of class a are (DTC) Dithiocarbamates Institute forEnvironmental Studies, Vrije Universiteit Amsterdam, The Netherlands 1 , J.Kamstra 1 , P. Cenijn 1 , J.Legler 1 286 357

Posters Posters 358 bge genephenocopiesthesemicircular canalprojectiondefectsseeninthemutant. the of function down knock to designed morpholino site splice A protein. the in residues acid genotyping assays. Both are missense mutations that predict the substitution of conserved amino were for mutations DNA genomic Point and cDNA system. in canal found semicircular developing the of projections epithelial the of Expression unknown. still is function their a candidate gene approach. Orthologous genes have been cloned in the human and mouse, but the Weidentified mutant. have the of in fuse to failure projections the the of result secondary a is swelling the that suggest data to correctly.Preliminary fuse fail and abnormalities expression gene and morphological show system canal semicircular become swollen at the end of the third day of embryogenesis. Epithelial projections forming the lauscher (lau) mutant is allelic to (bge) is an adult viable zebrafish mutation isolated in theWe Tübingen areusingzebrafishasamodelsystemforhumandeafnessandvestibulardisorders.bigears 1996 mutagenesis screen. The University ofSheffield, Sheffield, UK MRC Centre for Developmental and Biomedical Genetics and Department of F.Biomedical Geng,L. Science,Abbas, andT. T. Whitfield Development ofsemicircular canalsinthezebrafish innerear 287 bge by complementation assay. The inner ears of these mutants and bge appears to be ear-specific, and is restricted to restricted ear-specific, is be and to appears bge lau, and have been confirmed by PCR-based by confirmed been have and gene by genetic mapping and mapping genetic by gene bge Diseases models 1 C. Brusegan gene forParkinson’s disease Zebrafish as a model for neurodegenerative diseases: functional analysis of Diseases models gene at the PARK11 locus, however, further studies did not confirm its association with the with association its of confirm role causative the not elucidate the to did Therefore, disease. be studies further to however, locus, proposed PARK11 the been at recently gene has Protein-2) GYF (Grb10-Interacting GIGYF2 have dominant orrecessivePD.However, scans forsomeoftheseloci,thespecificgeneremainselusive. linkage whole-genome Family-based already rigidity.mapped 12 chromosomal loci (PARK1 bradykinesia, –PARK13 tremor, muscular ) responsible for rare forms of resting autosomal and to instability, leading neurons, postural (DA) dopaminergic of loss progressive and disease. It is characterized by depigmentation of Alzheimer’s disorder,after the neurodegenerative substantia common most nigra second (SN), the caused is by disease Parkinson’sthe selective University ofMilan–Italy recent geneticdatathatdonotconfirmGIGYF2asaPDgene. supporting zebrafish, in system DA the of differentiation proper the for necessary brain not is gigyf2 morphological drastic cause not did injection, alterations or loss of DA neurons during development. Therefore, our morpholino of means by experiments, Moreover, territories. non-neuroectodermal other in as well as system, that found We technique. hybridization situ mount whole by development during expression spatio-temporal its determined and gigyf2 akno Isiue Mln Italy; Milan, Institute, Parkinson 1 , A. Pistocchi 1 , A. Ghilardi 2 eatet f ilg ad eeis o Mdcl Sciences, Medical for Genetics and Biology of Department 1 , S.Goldwurm GIGYF2 is expressed in the embryo central nervous central embryo the in expressed is gigyf2 , we cloned the zebrafish homolog zebrafish the cloned we vivo, in 1 , S.Duga 2 , andL.DelGiacco 288 in vivo results suggest that loss of function of loss gigyf2 GIGYF2, a candidate 1

359 in

Posters Posters as avaluablecontributiontoclinicaldiagnosticsforRP predisposition. new candidates, so far not associated with RP, to broaden the spectrum of disease causing genes human RP and to elucidate the underlying pathomechanism. This model can be used to identify of aspects recapitulate to model excellent an is zebrafish that shows Takenstudy together,our zebrafish. of theanalyzedtranscripts. in phenotype RP-related a induce Using semi-quantitative RT-PCR, can we furthermore detected inefficiently spliced introns in several knock-down their that novel selected suggests of candidates characterization Preliminary pathomechanism. RP an with associated been not have far so which genes, mediator new potential identified we RP addition, In known genes. already disease represent in transcripts isdown-regulated identified the genes of Several morphants. expressed PRP31 of retina and PRP8 number significant a that suggest results Our retinae. deficient (OKR) PRP31 and PRP8 from orderderived cDNAs using assays microarray performed we In response deficits. opto-kinetic visual morphants, these in pre-mRNAs severe target the degraded ultimately exhibit and spliced inefficiently morphants identify measure to PRP31 to and PRP8 tests that suggest Behavioural furthermore layer. photoreceptor the of organization irregular and defects retina to leads embryos by zebrafish PRP31 into and injection morpholino PRP8 of knock-down Gene effect. specific tissue a mediating thereby mRNAs, cell-type to pre- lead expressed retina of subpopulation distinct factors a of processing affects primarily splicing that machinery general in splicing defects the of weakening general how a proposes hypothesis working elucidate, Our phenotypes. RP specific to is project our of Aim are corecomponentsofthetri-snRNP compleximplicatedinpre-mRNA processing. which PRP31, and PRP8 PRP3, in mutations include a and cases for RP of account percentage considerable These processes. cellular general encodes for required been genes genes also housekeeping RP have in knownidentified mutations RP of however, Surprisingly, majority photoreception. in The involved unknown. proteins remainsfor number still large genes a RP, but causing for cause disease as eye of identified been genetic have genes a 100 than (RP), more in pigmentosa Mutations Retinitis from suffers individuals disease characterized by the progressive 3500 degeneration of photoreceptor cells in in the neural retina. 1 Approximately Singapore 1 360 J. Brocher pigmentosa retinitis for model zebrafish a in degeneration photoreceptor to deficiency leads factor Splice f iceity Uiest o Wezug Germany; Wuerzburg, of University Biochemistry, of eatet f ilgcl cecs DS, ainl nvriy f Singapore: of University National (DBS), Sciences Biological of Department 289 1 , M.Graf 1 , B.Linder 2 , H.Dill 2 , U.Fischer 2 , G.P. Lee 3 eoe nttt o Snaoe (GIS), Singapore of Institute Genome 3 , S.Mathavan Diseases models 3 , C.Winkler 2 Department Department 1 ofGenetics, Japan;Shizuoka, Mishima active H-Rasexpression. constitutively widespread of presence the in transformation to pathway alternative an provides oncogene-induced Thus, arrest. cycle senescence of adult proliferating cells contributes cell to the development of Costello syndrome and p53-dependent and response of damage DNA Overexpression the of responses. damage DNA activated compartments with cell together H-RAS progenitor heart, adult and in brain markers the senescence in in increase an and proliferation oncogenic of levels low fish, Costello-like gene the in mutations activating by caused disease genetic rare a syndrome, Costello of hallmarks several display germline their in express high levels, of oncogenic to induced be can or levels, low express constitutively that zebrafish transgenic generated have pathway owing to germline mutations can cause severe Ras the developmental of Moreover,activation disorders. senescence. aberrant cellular of In inducer potent this a cancer,be study or human we in transformation and proliferation drive can H-RAS ‘oncogenic’ active, Constitutively Oncology, Milan, Italy for Genomic Technologies, Milan, Italy; Fagagna . Santoriello C. in adultproliferating cells Expression of H-RASV12 in a zebrafish model of Costello syndrome causes cellular senescence Diseases models 1 IFOM, the FIRC Institute for Molecular Oncology Foundation, Milan, Italy; through a heat-shock-inducible promoter in larvae led to hyperproliferation, activation hyperproliferation, to led larvae in promoter heat-shock-inducible a through 1 ,M.Mione 1 G Deflorian G. , 1 1 F Pezzimenti F. , H-RAS. We observed that fish carrying the integrated transgene 3 Division of Molecular and Developmental Biology National Institute H-RAS, 4 Department on Experimental Oncology, European Institute of of Institute Oncology, European Experimental on Department H-RAS 1,2 n cn e sd s mdl o te ies. In disease. the for model a as used be can and K Kawakami K. , expression are associated with both reduced both with associated are expression 3 L Lanfrancone L. , 290 2 Cogentech – Consortium 4 F dAd di d’Adda F. , 361

Posters Posters (MEMOSAD)) andHans-and-Ilse-Breuer-Foundation. Bayern, BMBF, EU Seventh Framework Programme (FP7/2007-2013,Fundedby: Deutsche-Forschungsgemeinschaft596), (SFB Bayern UniversitätElitenetzwerk and grant agreement no. 200611 model. for validated and design which inhibitor,drug rational GSK3β by active developed highly we novel a be identified We to progression. believed disease for is Tautrigger of a phosphorylation since GSK3β, Tau-Kinase the targeting compounds be can death cell neuronal that microscopy time-lapse by imaged time first the for demonstrate already could We approaches. treatment develop and of Tauopathies pathology transgenic the understand the better tools Therefore, important are size. fish small and transparency optical their to due development both drug for suited ideally are larvae zebrafish models contrast vertebrate In death. existing cell to and disturbances behavioral and neuronal as well as formation, tangle Tauprotein, human of changes conformational and phosphorylation Tauopathiesincluding of the have We tounderstand compounds. generated the firstdisease-modifying Tau-transgenicfor zebrafish, which rapidly models recapitulatescreen key pathological features and invaluable pathology are of mechanisms animals molecular modified Genetically available. are cures and hyperphosphorylation including lesions disorders subsequent aggregation of Tau neuropathological These protein and neuronal cell death. Currently, typical no mechanism-based Dementias. by Frontotemporal characterized certain are and disease Alzheimer’s include which Our ageing society is confronted with a dramatic increase of patients suffering from Tauopathies, Munich, Germany Molecular Biology,Germany;Max-Planck-Unit,Hamburg,Molecular 1 362 D. Paquet A zebrafish modelof Alzheimer’s Disease Biochemistry, LMU, Munich, Germany;Biochemistry,Munich,LMU, 291 1 , R.Bhat 2 , E.M.Mandelkow . Furthermore, we used our fish-model to identify new identify to fish-model our used vivowe in Furthermore, . 3 , RW. Koester 2 AstraZeneca R&D, Soedertalje, Sweden Soedertalje, R&D,AstraZeneca 4 , C.Haass activity in the transgenic fish transgenic the in activity vivo in 4 Neurogenetics, HelmholtzZentrum,Neurogenetics, 1 , B.Schmid Diseases models imaging and imaging vivo in 1 3 Structural & Structural Whitehead Instituteand G. DeRienzoandH.Sive DISC-1 isanessentialmodulatorofthewntpathway Diseases models implicated inhumanmentalhealthdisorders. of genes function the to explore atool as zebrafish using of efficacy the confirm pathway,and In conclusion, our data identify DISC-1 as an essential positive modulator of the canonical Wnt zebrafish development. during GSKβ modulating in DISC-1 of role fundamental the demonstrating DISC-1 human with the DISC-1 loss of function phenotype. Finally we were able to rescue the morphant phenotype increasing Fourth, function. of loss DISC-1 after expression gene endogenous and reporter rescue to able is inhibitor GSK3β a function DISC-1 of loss Second, function. decreased Wnt8b reporter gene of indicate expression loss in a or Wnt-responsive evidence DISC-1 reporter zebrafishof line,loss TOPdGFP. of after observed Third, expression are lines gene forebrain in changes additional similar function First, pathway. Wnt Several the of in functions phenotype, genes. DISC-1 loss that strong a these Wnt8b in between resulted and each synergy of DISC-1 function demonstrating of from loss partial resulting that showed phenotypes and similar, gross are the that noticed We DISC-1 somitogenesis, normal characterized by a massive deficit inall neuronal outgrowth and poor fasciculation. ablates and 1/2, intron and early by apparent phenotype, strong very a show (“morphants”) embryos injected D1MO RNA. 1 exon D1MO wasdirected between site donor The splice sites. the against splice targeting (MOs) oligos antisense morpholino-modified order to test the embryonic function of DISC-1, we decreased zygotic gene function In by injecting system. nervous central developing the maternal in expressed shows strongly is and DISC-1 expression zygotic Zebrafish and mammals. and zebrafish in domains gene DISC-1 conserved the highly in similarity,present acid are amino overall low Despite phenotype. human the animal an as “tool” a define system Wethat offers GSK3β. useful with insight into interacting function of by a disorder schizophrenia risk of gene, but etiology that may the not in recapitulate We have begun to use the zebrafish as a tool to study the function of DISC-1, a gene implicated 1 Massachusetts Instiuteof Technology, Cambridge, USA 1 ctnn ees truh n nuil fso poen rescues protein, fusion inducible an through levels, β-catenin 292 363

Posters Posters 364 by differentmutations. mutations and have implications for future discriminatory therapy of this human disease caused a in decay (NMD). Our zebrafish possibly studies thus provide novel insight into degeneration the pathomechanism mRNA of nonsense-mediated both to due haploinsufficiency rod by degeneration rod causes and unstable in results protein mutant dominant-negative fashion, whereas mutant SP117 protein is AD5 mislocalized to the cytoplasm and of Expression mechanisms: expressing rhodopsin of number the our Hence, change cells. not did PRP31 while wildtype rodphotoreceptors, or SP117 expressing of of rhodopsin expression reduction significant a in AD5 resulted the of mutant expression transient that found we addition, In protein. mutant the of localization majority of mCherry signal was detected in the cytoplasm of rods indicating aberrant subcellular situation in AD5:mCherry injected embryos. In contrast, in SP117:mCherry injected embryos, the wildtype with mutant or wildtype encoding plasmids PRP31 fused in with frame to mCherry rods. injected under the control of of then the rhodopsin cytoplasm promoter. were Embryos the injected embryos in transgenic GFP rho-GFP expresses which line, transgenic (rho-GFP) rhodopsin-EGFP a expression at 14hpf. For an variant had a similar stability as injected wildtype AD5 the 11hpf,whereas at detectable longer After no was and degeneration. quickly degraded cell SP117protein the photoreceptor to lead aa), that found we embryos, zebrafish into variants 499 PRP31 mutant both encoding RNAs of injection wildtype aa; 469 and (277 proteins cause a frameshift at amino acids 256 and 371 respectively which and might result in AD5), truncated PRP31 (SP117and mutations human two mechanisms which by investigate to diseases, vitro in mechanisms the study underlying to RP caused used by PRP31 mutations been and general conflictinghave results have beenmodels obtained from the culture such cell far,mostly in So humans. mutations in RP of Different form autosomal-dominant cones. the for cause of major a as loss identified been have the PRP31 factor splicing in results secondarily and rods firstly which affects retina, the in cells photoreceptor of degeneration progressive a by characterized Retinitis pigmentosa (RP) is an inherited eye disease with a prevalence of approximately 1/3500, ofBiological Sciences(DBS),Department NationalUniversity ofSingapore J. Yin, C.Winkler in azebrafish modelforRetinitispigmentosa PRP31-AD5, and PRP31-SP117 mutations, disease human two of characterization Functional 293 approaches. We have used transgenic zebrafish as a powerful animal model for human for model animal powerful a as zebrafish Wetransgenic approaches. used have PRP31:mCherry showed transient expression in the nuclei of rods, similar to the to similar rods, of nuclei the in expression transient showed PRP31:mCherry data suggest that the two analyzed PRP31 mutations act by different by act mutations PRP31 analyzed two the that suggest data vivo in in vivo assay of protein localization and function, we next generated PRP31 and showed continuous high levels of Diseases models a scientific basis for clinical use of the selected CM and the identification ofnewpotential provide and theidentification disorders. to CM modelfor expected are theselected ofzebrafish of research use compounds leading this of application to clinical safe of and for effective basis study results drug scientific development first The for a the the treatment CM. of is neurodegenerative from this screening knowledge, neuroprotectants our of best the To AOE neededtobeelucidated. the in compounds different of interaction synergistic and metabolites study,active future the In zebrafish. in activity for properties pharmacokinetic favorable and conservation functional with flavonoid (a chrysin neuroprotective in mild zebrafish namely showed model,AOE, butfrom not inisolated compounds, PC12acid, cellprotocatechuic model. Itand pure mayquercetin) beto duestructure-similar towith these two chemicals However, manner. dose-dependent zebrafish by in apoptosis neuronal LGA-induced prevent notably could AOE and quercetin both Moreover,zebrafish. in loss neuron dopaminergic induced MPTP and 6-OHDA and death cell Oxyphyllae (AOE), exhibited significant neuroprotective activity against 6-OHDA induced PC12 of extract ethanol activity, and anti-oxidative with flavonoid well-known a quercetin, damage in both cell culture and zebrafish models. Among these compounds and extracts tested, and non-specific oxidative agents (L-2neuron hydroxyglutaric dopaminergicacid, LGA) were used to induce against neuronal cell MPTP) and (6-OHDA neurotoxin selected study, present In andzebrafishinvivo.activities byacombinedmodelofPC12cellinvitro neurodegenerative of treatment for 14 known pure societies compounds from CM and Asia 2 herbal extracts were in investigated for neuroprotective application of disorders. In history an effort long to find effective a neuroprotective ingredients has from Chinese(CM) medicine, herein, increasing severe public represent health sclerosis issues, multiple and which alzheimer’s parkinson’s, is as such lack disorders of Neurodegenerative effective treatment so far. Chinese medicine Institute ofChineseMedicalSciences, University ofMacau, Taipa, MacaoSAR, China Z. Zhang,S.M-Y. Lee,,M.Wang, A. Deepa Fishing forneurorpotectantsfromChinesemedicine Diseases models 294 Allpinae 365

Posters Posters (TBH). The two ruthenium compounds were less toxic than SDS and CuSO activity and the toxicological parameters studied in zebrafish embryos that are related to the to related are that same timetotheirstructures. embryos zebrafish in studied parameters toxicological the and activity model,the animal the the between correlation in an and reliable a complexes present here ruthenium evaluated precursor the ruthenium of behavior complexes the Ru(II)-NHC that showed testing results The study embryo. zebrafish first the is this conclusion, In the highestconcentration,whereaminordecreaseinheart ratewasdetected. The results. for except alteration viability no showing precursor ruthenium the again was substance evaluated confirming safest sulfate, copper the by followed SDS, toxic the most was the compound that indicate results The compounds. reference the to comparison in analyzed was rate heart the in treatments compound ruthenium the of influence the and well, as studied treated with the ruthenium precursor that remained inside of the chorion. The cardiotoxicity was the totality of incubated embryos with remained the compounds died, except the controls and compounds the animals ruthenium concentration, highest the the At development. egg the in delay with a suggesting chorion, the of inside treated embryos the However, dead. were sulfate copper or SDS with incubated animals the while controls, to comparison in differences present not did TBH and precursor ruthenium organs.the with Withtreated embryos the dose, higher a their of coagulation by coloration opaque presenting dead result SDS with treated animals the (RuCl pro-oxidant dose-dependent activity at the highest concentration, whereas a ruthenium precursor the In model. zebrafish the vitro antioxidant/pro-oxidant activity of a series of Ru(II)-NHC complexes and their toxicity using improved and no and data are available in the new literature on animal models. In of this work, we have searchstudied the the metallodrugs. in However, chemists only little organometallic data with and regard gathering to biomolecular interest, their toxicity great biomedical, have attract been to available beginning until are now, complexes metal NHC-based of applications have demonstrated their capability to support a wide variety of metals. Recently, the biomedical ligands (NHC) carbene N-heterocyclic sense, this In complex. the of features final the defining role, important an play ligands ancillary the centre, metal the Besides drugs. metal-based new of design the in potential great a demonstrated have complexes ruthenium years, last the Over 366 a Poyatos M. Oxidant VitroIn Activities andIn on VivoBased Evaluation ofBiosafetyinZebrafish EmbryosComplexes Ruthenium-NHC of Series a of Properties Biomedical Ripoll-Gómez the same concentration the embryos treated with CuSO with treated embryos the concentration same the controls, However,at hpf. the 72 at chorions with their of out being differences and body developed present well a clearly showing not did TBH with or precursor ruthenium compounds, ruthenium with incubated to embryos the dose, due studied lowest the embryos At studied. were treatment in the changes morphological Additionally, series. the of compound safest the is precursor did not present loss of viability at the maximum dose used, indicating that the precursor b n oprsn ih he rfrne opud: D, CuSO SDS, compounds: reference three with comparison in embryotoxicity test, the two ruthenium compounds and the ruthenium precursor were analyzed Drug Discovery Unit, Neuron BPh, Parque Tecnológico de Ciencias de la Salud, Granada, Spain Dpto. yOrgánica, deQuímicaInorgánica Universitat JaumeI, Castellón, Spain 295 2 (dmso) b , J. S. Burgos S. J. , 4 dd o so ay r-xdn bhvo a ay tde ds. n h zebrafish the In dose. studied any at behavior pro-oxidant any show not did ) a

a , J. M. Alfaro, M. J. , assay, vitro in significant showed compounds Ru(II)-NHC the of two a A. Prades, A. b M. del Carmen Ramos, Carmen del M. 4 remained inside of the chorion, while chorion, the of inside remained 4 n tr-uy hydroperoxide tert-Butyl and Diseases models 4 , while the ruthenium a E. Peris, E. oxidant vitro in b and J. and in

that we are validating in zebrafish. Zebrafish thus provide a useful model for validating the forvalidating model useful a provide thus pathogenic natureofhumanbraindisease-relatedalleles. Zebrafish zebrafish. in validating are we as that aswell inherited several identified have we project rescued be genomics could our For approach. this alleles, of phenotype usefulness disease-related the establishing the thus the mutations, of novel including some tested, by not cases but mRNA human the native the In of overexpression projections. by axon neuralpopulation in spinal changes in led to defects or motility, abnormal included which phenotypes specific yielded genes homologues zebrafish the of knockdown that Wefound paraplegia. have spastic sclerosis, include tested genes The diseases. related cord genes spinal in human mutations to several studied successfully we approach experimental this Using the disease-relatedsequencevariants. of effectiveness the compare we observed, is rescue If morpholino. the with co-injection upon mRNA human native the expressing by phenotype knockdown the rescue to attempt next We database and designing a morpholino antisense oligonucleotide to selectively target translation. disease-related possible to comparison in variants. and For each human gene, gene we start by identifying human the zebrafish homolog fromnative then the genomic we the which with phenotype rescue a to yield attempt to gene zebrafish homologous the of expression initial the down the However, their of validation schizophrenia. biological pathogenic lacks nature. often We and disease are human therefore with using we to autism zebrafish related patients to screen mutations Montréal, potential of in gene including identification variants de by genes knocking disorders, synaptic Université of brain the mutations at developmental for groups screening research genomics large-scale genetics performed human with collaboration In ofPathologyDepartment andCell Biology, Université deMontréal, Canada P. Drapeau, E.KabashiN.Champagne,Brustein,M.Liao,C.Maios,Lapointe Validation ofmutationsrelatedtodevelopmental humanbrain diseasesusingzebrafish Diseases models linked to skin and spinal cord disorders and disorders cord spinal and skin to linked AP1S1 uain o snpi genes synaptic of mutations novo de related to amyotrophic lateral amyotrophic to related ALS2 associated with hereditary with associated SPG8 296 367

Posters Posters 368 and theProgram Research forNeurology &Discovery. MN of (SAS:NIH the by course NS007222-26), Supported the A. Taubman Alfred Institute, the Research Medical throughout intervention therapeutic for degeneration. points identifying by ALS treat to the from obtained Knowledge understand ALS. to with characterization of order stable transgenic zebrafish as associated a model of in ALS will greatly degeneration advance our ability survival axonal and of morphology mechanisms axonal the integrity, NMJ analyzing by ALS of model ability.this swimming characterizing of Wecurrently loss are progressive a demonstrate stable ALS, for treatments G93A-SOD1 novel transgenic zebrafishidentify have beenand generated. neurodegeneration Theseof G93A-SOD1 transgenicmechanisms zebrafish the study To mutation associated with familial ALS, results in defects in MN outgrowth and axonal branching. and underlie that neuroprotection. Transientmechanisms as well genetic axons manipulation as of zebrafishprogression, to expressand G93A-SOD1, a onset MN ALS in events of early study to system observations vivo in Detailed excellent MNs. an provide they therefore, of zebrafish; in possible are loss (NMJs) junctions neuromuscular irreversible the to prior therapeutic intervention for target appealing an it making disease), MN (ALS; sclerosis lateral amyotrophic Axonal degeneration precedes symptom onset and motor neuron (MN) cell death associated with University ofMichigan, Ann Arbor, USA S. A. Sakowski, A. S.Busta,J.Dowling,E.L.Feldman G93A-SOD1 transgenic zebrafish asamodelofamyotrophic lateral sclerosis 297 Diseases models ameliorating amyloidtoxicity. model to display an earlier phenotype. This could prove useful for analysis of drugs intended to transgenic this modifying currently are We melanocytes. of loss progressive showed fish adult Aβ human of form the residue using acid melanocytes amino in 42 specifically the expressing by zebrafish in toxicity amyloid of in changes observe to used be can that substrate APP modified a produce that zebrafish transgenic of development begun no is there Currently invading by manner of splicing aberrant the that hypothesise We effect. negative dominant a have truncations particular embryos, into encoding mRNA synthesized have of splicing aberrant that dominant negative effects on the function of of disruption that demonstrated Wehave 1) Results: orthologues ofhumanPSEN1and2,APP. possesses it as of pathology bases AD molecular the investigating for model vertebrate effective PRESENILIN 1 catalytic cores of clearance that appears to promote neuronal dysfunction and death. Presenilin proteins form the Aberrant (APP). protein precursor lesions Aβ the proteolytic processing of of APP observed by fragment cleavage have a is peptide patients Aβ (NTF). tangles AD of analyses neurofibrillary intracellular and (Aβ) peptides Histological amyloid-beta of deposits extracellular containing elusive. remains (AD) disease Introduction: An integrated understanding of the exact molecular and cellular basis of Alzheimer’s Molecular &BiomedicalScience, The University of Adelaide, Adelaide, Australia L. Wilson, S.Nornes,M.NewmanandLardelli presenilin, investigate research to zebrafish Using Diseases models and 2 γ-secretase complexes. Genetic studies have discovered mutations in that contribute to familial autosomal dominant AD. (above) produces truncated proteins that act in a dominant negative dominant a in act that proteins truncated produces (above) PSEN sceae opee t peet h frain f h ctltc ie 3) site. catalytic the of formation the prevent to complexes γ-secretase assay appropriate for investigating for appropriate assay vivo in in ageing neural cells may contribute to sporadic AD. 2) We 2) AD. sporadic to contribute may cells neural ageing in PSEN1 -secretase activity.γ-secretase model Wetransgenic 4) a develop to attempted γ-secretase results in an imbalance between Aβ production and rnain atr xn ucin. hn injected When junctions. exon after truncations PSEN1 rmtr Fo 1 mnh o ae oe f these of some age of months 16 From promoter. mitfa PSEN1 and the related gene sceae ad P fr lhie’ disease Alzheimer’s for APP and γ-secretase, transcript splicing can have potent have can splicing transcript PSEN1 -secretase activity. We have Weactivity. γ-secretase 298 PSEN2. We hypothesise Danio rerio provides an APP and 369

Posters Posters 370 kidney malformed causes consequently and expression, wt1b of phenotypes. regulation down to leads APAPwith treatment that suggest Takentogether,we domains. expression wt1b reductive had and durations dose, exposure APAPafter embryos revealed treatment hybridization situ in whole-mount Furthermore, onsets. the on depended nephrotoxicity APAP-induced that indicated is longer then 24h 6800ppm APAP, no surviving embryos were observed by 72hpf. These results higher displayed embryos duration exposure the When 60-72hpf). (ex: whereas onset exposure later of at defects percentagesmild of 12-24hpf), percentages (ex: exposure higher of displayed onset earlier embryos at defects (3400ppm), severe dose and 12h) (ex: same duration the under exposure Specially, increased. APAP of dosages kidney exposure malformed the with as embryos increased of morderate phenotypes percentages to The similar midline. by defects: separated severe are (3) defects gl position; mild but normal defects defects: at gl morderate atrophic (2) and (gl); cystic a glomerulus with normal but the a and with (pt) tubule but pronephric (pd) cystic duct and pronephric curved defects: mild (1) groups: three as classified dose (3400, 6800ppm) treatment, embryos displayed malformed kidney phenotypes which were the in changes morphological vehicle-control (0ppm) and and 340ppm rates APAP-exposure (12-72hpf) survival groups. In in constrast, after higher differences evident no exhibited embryos We dynamically. durations (12, 24, 36, 48, 60h) nephrotoxicity and onsets (12, 24, 36, 48, 60hpf). Results showed APAP-inducedthat zebrafish the of observe effects carried to out a toxic series model of exposure experiments the line, a with kidney different concentrations but fluorescenct as (340, green 3400, 6800ppm), damages a Tg(wt1b:eGFP), used kidney we Here, and embryogenesis. during In liver known little decades. to is APAP for leads drug APAP antipyretic of and over-dosed analgesic an mammals, as used widely is (APAP) Acetaminophen Institute ofLifeScience, Tamkang University, Taipei County, Taiwan, R.O.C. H-C. Peng, Y-H. Chen Acetaminophen-Induced NephrotoxicityinZebrafish 299 Diseases models compromise neuromuscular transmission in CMS patients. Furthermore, zebrafish may be a may zebrafish Furthermore, suitable modelorganism fortestingnoveltreatmentspatientswithDOK7mutations. patients. CMS in transmission neuromuscular compromise which through mechanisms molecular the determine to help may results Our role ofDok-7instabilisingandbranchingNMJs. a imply might results These embryos. deficient Dok-7 in affected be to seem not do formation extended have embryos wild-type branches in into and formed axons synapses with laterally motor located muscle fibres.time, this Very early stagesBy of NMJ (hpf). post-fertilization 48h at into fertilised zebrafish eggs revealed first abnormalities of NMJ patterning in zebrafish embryos oligonucleotide morpholino antisense an of injection by expression Dok-7 of Downregulation the in studied be endplate maintenance inazebrafishmodel. can and development development endplate in of Dok-7 of stages role the early investigated therefore at We zebrafish. mammals in lethal are that mutations vertebrate development and is also receiving increasing attention as a model of human disease; mice were immobile at birth and died shortly thereafter. The zebrafish is an established model of However,vivo. in role deficient its Dok-7 explore to Dok-7 lacking mice generated co-workers and Okada yet. elucidated, been not has NMJ the at protein Dok-7 the of function precise The patients withDOK7mutationsdonotbenefitfromlong-termtherapyesteraseinhibitors. accounting for about 10% of genetically diagnosed CMS cases. In contrast myasthenia. to of other CMS subtypes, gene, CMS novel synaptic a in Recently, mutations terminal, identified. been have nerve proteins apparatus postsynaptic presynaptic and cleft, in mutations Causal (NMJ). junction neuromuscular at the transmission affect that defects genetic from arise (CMS) syndromes myasthenic Congenital Institute ofHumanGenetics, Newcastle University, Newcastle upon Tyne, UK J. S.Müller,H.Laval,K.Bushby, V. StraubandH.Lochmüller Neuromuscular junctionformationinDok-7deficientzebrafish embryos Diseases models (“downstream of kinase”) 7, were found to cause an autosomal recessive form recessive autosomal an cause to found were 7, kinase”) of DOK(“downstream uain hv eegd s n o te ao gntc eet i CMS, in defects genetic major the of one as emerged have mutations DOK7 300 mutations DOK7 371

Posters Posters 372 L. Araujo-Bazan model photoprotective for a zebrafishscreening vertebrate as using New compounds 1 of photoxidativestressandphotodamage. Our results confirm the use of willbe zebrafish model to determine the effects of conditions UVB radiation in terms theexposure presented. underlying the mechanisms and Arrays’) Genome Zebrafish ‘Genechip (Affymetrix: microarrays cDNA using by studying being are pathways biochemical Vitaminwith treated embryos irradiated in C. observed Affected was effect photoprotective less apoptotic cells percentage of the PL and Trolox treated irradiated embryos were observed. 48 Much to up Acridine Orange vital fluorescent viability dye. A significant and increase inassessed the viability abnormalities was and aphotodamage decreaseDNA in morphological the of Evaluation the hpf. evaluating by compared untreated were compounds those different with the with treated embryos Irradiated concentrations compounds. different the with of incubation previous without or with were radiation, hpf) UVB (24 to embryos exposed Zebrafish C). (Vitamin acid L-Ascorbic and (Trolox) acid 2-carboxylic (±)-6-Hydroxy-2,5,7,8-tetramethylchromane- like properties, photoprotective and other antioxidant and extracts toperform easily natural Polypodium and leucotomos of (PL) has been investigated besides fast another compounds, properties known for their new photoprotective a fern the of extract hydrophilic a of effect and photoprotective The developed. been has compounds antioxidant model, embryo detect zebrafish to the method using study, present the In and easinesstohandleinmultiwellplates. toxicology assays and screening of therapeutic agents, due to its transparency, fast development In recent years, zebrafish (Danio ) rerio has increased its importance as a organism. vertebrate model both for the reflect not does conditions culture as under cells of response the such effects deleterious to models have leads been mostly used to perform (UVR) quantitative studies of radiation cytotoxicity; radiation however, cancer.skin ultraviolet and photoaging suppression, culture immune Cells sunburn, stress, photooxidative solar to overexposure Skin Sloan-Kettering CancerCenter, New York, USA ZFBiolabs, Tres Cantos, Madrid, Spain. Madrid, Cantos, TresZFBiolabs, 301 1 , J.Guinea 1 , E.Reyes 2 , S.Gonzalez 2 IFC, Madrid, Spain. Madrid, IFC, 3 andI.Rodriguez-Martin by quantification of apoptotic cells using cells apoptotic of quantification by 3 Dermatology Service, Memorial Service, Dermatology response of a whole a of response vivo in Diseases models 1 in vitro/in vivovitro/in in 1 1 The roleofthetumoursuppressorLKB1indevelopment andcancer Diseases models lkb1/+ fishtoestablishwhetherLKB1actsasatumorsuppressoralsoinfish. aged adult of analysis the started recently we addition, the In at level. transcriptional mutants and lkb1 molecular in regulated aberrant are pathways which investigating are we Currently, that byhistochemistry of thelarvaeat8dpf. shown have we finding, mutants display defects in this energy post metabolism control days with resulting 11in premature around starvation Consistent and at death larvae (dpf). wild-type starved fertilization of those resembles rather but affected not is cells epithelial intestinal of polarization cell surprisingly, Perhaps epithelium. the of flattening observe we completed, is the absorption of yolk emaciation when onwards, 5 day From development. of days ina Homozygous result Both mutations byTILLING. ortholog premature stopcodonwithinthekinasedomain. LKB1 zebrafish single the in mutations two identified have cancer,we and gain development Toduring polarity.effects its cell mediates LKB1 how with into insight associated including is pathways LKB1 Additionally, several pathway. mTOR through the suppression of growth inhibition to leads that and conditions energy low under AMPK activates LKB1 tract. gastro-intestinal the of a particular in origin, syndrome, epithelial of Peutz-Jeghers causes LKB1 kinase gastrointestinal serine-threonine hamartomatous polyposis disorder with the increased predisposition to in malignancies mutations Germline Institute, Utrecht, The Netherlands oeua Gntc, ehrad Cne Isiue Asedm Te Netherlands The Amsterdam, Institute, Cancer Netherlands Genetics, Molecular YU. van dervelden mutants are morphologically indistinguishable from wt siblings the first 5 first the siblings wt from indistinguishable morphologically are mutants lkb1 mutants concurrent with an abrupt collapse of the intestinal villi and villi intestinal the of collapse abrupt an with concurrent mutants lkb1 1 M. vanLohuizen 2 H. Clevers 1 AP. Haramis 302 2 Hubrecht Hubrecht lkb1 373

Posters Posters asrct Te Netherlands; The Maastricht, Belgique; (GIGA),appliquée Génoprotéomique Liège,de de Universite (LBMGG); Interfacultaire Grappe H.J.M. Smeets 374 Bosch den van B.J.C. Zebrafish asanewmodeltostudy mitochondrial disease is critical. to common and age-related diseases in which mtDNA copy number and mitochondrial function suited for this. Moreover, this does not only apply to the severe neurological can syndromes, but also that compounds of reduce or identification delay exists, pathology is disorders of major OXPHOS importance. The for established zebrafishtreatment models areeffective perfectly no As largely the unravel to phenotypes. associated the value and defects mtDNAreplication in processes great pathological unknown of are models these established, mtDNA When with defects. patients in replication affected often most are and production energy their for OXPHOS on In patients. in manifestation rely mainly organs disease these because investigated, be will liver on and muscle particular,brain, heart, based determined be and Phenotypes will mice. parameters and yeast mitochondrial in damage catalysis, oxidative to polymerase leads and lowers errors Toreplication drastically induces organism. mutation expressed be This whole will transgenesis. the mutation transposon-mediated Y955C for dominant-negative by the approach model, dependent POLG transgenic dosage a gene create a established allowing be will morpholinos deficiency POLG by of model embryonic An deficiencies. OXPHOS to lead patients. Our aim is to develop a zebrafish model to investigate how mtDNA ophthalmoplegia replication defects OXPHOS all of external 25% about for progressive account possibly mutations thrive, POLG failure. liver and to epilepsy to failure from ranging humans, in phenotypes mitochondrial of range in broad a cause mutations Defects POLG and crucial is (POLG) diabetes. gamma polymerase replicating 2 type and mtDNAThe conditions. diseases these in factors key are mtDNAreplication particularly and biogenesis cardiac like disorders, common in involved alos are but syndromes, neurological onset early rare tomany to lead fundamental Deficiencies life. is (OXPHOS) phosphorylation oxidative by production energy Mitochondrial Netherlands Netherlands; 1 eatet f lncl eois Matih Uiest Mdcl ete Matih, The Maastricht, Centre, Medical University Maastricht Genomics, Clinical of Department 303 4 Department of Neurology, Erasmus MC University Medical Center,The Rotterdam,Medical University MC Neurology,Erasmus of Department 1,2 2 eatet f lncl eeis Matih Uiest Mdcl Centre, Medical University Maastricht Genetics, Clinical of Department 1,2 M Müller M. , 3 aoaor d Booi Mlclie t e ei Genetique Genie de et Moleculaire Biologie de Laboratoire 3 E Jongen E. , 2 ATM Hendrickx A.T.M. , Diseases models 2 IFM d Coo de I.F.M. , 4 , es tre osiuie RA pie factors, splice mRNA constitutive three least at in mutations However, photoreception. in involved proteins affect mutations RP-causing of majority The cells. photoreceptor cone of loss secondary with degeneration photoreceptor rod Retinitis Pigmentosa (RP), an inherited disease leading to blindness, is characterized by progressive etr abr-pedr, Germany; Hamburg-Eppendorf, Center C. Winkler 1 . Dill H. Analysis ofdiseaseassociatedsplicefactorsinazebrafish modelforretinitispigmentosa Diseases models Sciences, National University of Singapore; Germany; activity causeRP byinfluencingsplicingofretinaspecifictranscripts. spliceosomal the affecting mutations that idea the Togetherwith consistent genes. are data our housekeeping to compared down-regulated, significantly be to out turned genes retina-specific of analysis microarray In larvae. zebrafish that indicate tests RP of in features main the mimic could we Thus, cognition. visual in deficits exhibit morphants Behavioural normally. develops apparently embryo or of knockdown gene mild system, zebrafish the In degeneration. specific photoreceptor the mechanism, by which defects in ubiquitously expressed pre-mRNA processing factors cause tri-snRNP defective revealed hPrp4 mutant association. Furthermore we have established a ofzebrafish knockdown model to gain insight into analysis Biochemical gene. candidate RP new identified we factors, splice in mutations new carrying patients RP for screening machinery.By splicing pre-mRNA the of part essential an tri-snRNPparticle, U4/U6.U5 RP.so-called to the lead of that components are factors These Deptartment of Biochemistry, University of Wuerzburg, Germany;Wuerzburg, of University Biochemistry, of Deptartment leads to severely reduced expression levels of rhodopsin, even though the remaining the though even rhodopsin, of levels expression reduced severely to leads zfPrp4 1 B Linder B. , 5 2 Genome InstituteofSingapore, Singapore , B.Laggerbauer 1 A Hirmer A. , 1 and U.Fischer 1 J Brocher J. , 4 nttt f ua Gntc, nvriy f Cologne, of University Genetics, Human of Institut -morphant retinae, a considerable number of number considerable a zfPrp31retinae, -morphant , another well known tri-snRNP protein as a as protein tri-snRNP known well another hPrpf4, 3 1 Department Department of Human Genetics, University Medical 2

S G P Lee P. G. S. , hPrp3, and hPrp8 5 S Mathavan S. , ae en described been have hPrp31 304 2 Department of Biological of Department 5 and zfPrp31 A Gal A. , 3 H Bolz H. , zfPrp31 zfPrp4 375 4 ,

Posters Posters n re t caatrz i mr dti te fet o MC2 nc-on uig zebrafish during knock-down MECP2 the nervoussystemandneuromuscularsynapseformation. of effects the detail more embryonic development we are currently analyzing the cytological and histological structure in of characterize to order In pathfinding, suggestingafunctionforzMECP2interminalneuronaldifferentiation. zMECP2, in neuronalmaturationandmaintenanceasdescribedmammals. After thelossoffunction pax2a) wint1, (otx3, suggests that marker zMECP2 is not very essential to neural early brain development, but it could be with necessary morphants of analysis otic preliminary A and inflated. appear system nervous cephalic both of vesicles. reduction Midbrain-hindbrain boundary strong (MHB) is a thin, gene and showed both midbrain morphants and knockdown Moreover, hindbrain ventricles defects. motility to touch-provoked and spontaneous display (MECP2MO) embryos injected zMECP2MO fact, oligonucleotide RTT.In in phenotypes morpholino resembling defects behavioral and morphological antisense observed activity,we specific a Using and inadultbraineyes. high levels apowerful of its expression were found in embryos at 1 to 4 h as postfertilization (hpf), after 24 hpf, zebrafish 8 group propose linkage to mapped previously we obtained, were vertebrate models system to investigate the RTTrole of MeCP2 during embryonic murine development. zMECP2 was several Although RNA splicing. 2 (MeCP2), a transcriptional repressor involved in chromatin remodeling and the modulation of protein binding methyl-CpG encoding gene the in RTTmutation classic a of have 80% patients and purposeful hand speech movements and reduced in brain growth. disability,reduction It is mental now known profound that approximately a produces disorder this age), of foetal months 6-18 causing and births female live 1/10,000 to demise in up males. After an of early period of frequency apparently normal a or almost normal with development (until occurring females, Rett syndrome (RTT) is a X-linked dominant neurological disorder that affects almost exclusively Structural andFunctional Biology, Italy 1 376 G. Gaudenzi Zebrafish, anewmodeltostudy rettsyndrome nvriy f ia, eatet f ilg, Italy; Biology, of Department Milan, of University 305 we detected that neuromuscular axons of morphants can display slight problem in the 1 , A. Ghilardi 1 , A. Guarda 2 and it is highly similar to mammalian MECP2. Relatively MECP2. mammalian to similar highly is it and , F. Cotelli 1 , G.Badaracco 2 nvriy f nura Dprmn of Department Insubria, of University 2 Diseases models currently nootheranimalmodelsavailable. in understanding the pathomechanism leading to FTLD-U in humans, especially since there are gain- and loss-of-function zebrafish will be presented. These zebrafish models are valuable tools TDP-43 the of Characterization TDP-43. of mutants different or TDP-43 wildtype human either expressing lines zebrafish transgenic several generated we death cell neuronal and aggregation induce to sufficient is TDP-43 Toof gain-of-function gripNAinjections. if antisense investigate widely to are get further insight proteins into its physiological function, Both we knocked (TARDBPL).down TARDBP Like and TARDBPLexpressed during Protein by early development. To Binding Tar-DNA test and the hypothesis (TARDBP)if TDP-43 loss-of-function Protein protein, Tar-DNAis Binding TDP-43 toxic human and the to homologues close two are there the zebrafish In chose we possibilities these between distinguish zebrafish togenerateananimalmodelrecapitulatingaspectsof‘TDP-43Proteinopathies’. To toxicity. in results aggregates the to intermediates confer oligomeric toxicity or aggregates to the Either FTLD-U. the of pathogenesis neurons, the to contributes or aggregation 43 alternatively the loss of TDP-43 due to Lobar Degeneration sequestration with ubiquitin positive inclusions (FTLD-U). So far it Frontotemporal of is cases sporadic not and familial in known aggregated be how to species TDP- protein major the be under the age of 65. The patients Tar-DNAin dementia of binding protein form of 43 common kDa (TDP-43) was most recently identifiedsecond to the is Degeneration Lobar Frontotemporal Ludwig-Maximilians-University, Munich, Germany Butenandt-Institute, Adolf & (DZNE) Erkrankungen Neurodegenerative für Zentrum Deutsches B. Schmid, A. Hruscha,M.Teucke, D.Dormann,Paquet,F. vanBebber, C.Haass Modeling Frontotemporal LobarDegeneration inzebrafish Diseases models 306 377

Posters Posters 378 that activity.transport glucose in involved motifs RT-PCR revealed tissues zebrafish adult in analysis 12 typical zebrafish The acids. amino 513 of polypeptide a encodes bp) (1.542 frame reading open consensus The comparable to that of the human counterpart, consisting of at five exons separated by four introns. embryos and fishes his entire coding adult sequence. The gene, located from of on organization chromosome genomic 11, cDNA the define has to an exons/introns us of structure allowed stages, developmental sequencing different and analyses Computational . rerio the for searched have We unknown. still ATSis of facilitative glucose transporter type 10 (GLUT10), the role of this transporter in the pathogenesis recently andreported to be due to loss-of function mutations in the skin wrinkled hyperextensible soft, i.e., manifestations, tissue connective and features dysmorphyc characteristic Other include stenosis. signs arteries typical pulmonary and tortuosity,arteries, middle-sized elongation, and large with the of involvement aneurysms and arterial widespread by connective characterized recessive disease, autosomal tissue rare a is (OMIM#208050) (ATS) syndrome tortuosity Arterial Biotechnology, and Sciences Biomedical of Department Genetics, University ofBrescia, Italy and Biology of Division N. Chiarelli,M.Ritelli,Zoppi, A. Benini,G.Borsani,S.Barlati,M.Colombi in arterialtortuositysyndrome Characterization and expression of suggest thatglut10shouldplayaroleinvasculardevelopmentzebrafish. vesicle; in 4-day-old in vesicle; to 1-day-old embryos. In 48 hpf embryos, a distinct hybridization signal was detected in the otic up remained and notochord caudal the in observed was expression the somites) (20-25 hpf 19 at whereas notochord, the to and placode otic the to restrict be to appeared signal the somites) the stages. larval early the to onset segmentation the from decreases and stages embryonic early the in and zygote the in already present is transcript the fin. caudal 307 expression is ubiquitous until mid-somitogenesis. However, at 13.5-14 hpf (8-10 hpf 13.5-14 at However, mid-somitogenesis. until ubiquitous is expression slc2a10 gene is expressed in liver,in expressed is gene slc2a10 protein shares a 45% sequence identity with the human protein, human the with identity sequence 45% a shares protein glut10 domains structure and shares the signature sequence signature the shares and structure domains hydrophobic transmembrane Quantitative real-time PCR analysis showed analysis PCR real-time Quantitative the larvae transcript was present in the cardiac region. These data These region. cardiac the in present was transcript slc2a10 slc2a10, the zebrafish ortholog of the human gene involved heart, gills, eyes, brain, spleen, swim bladder,swim spleen, brain, eyes, gills, heart, and ovary Atog AS a been has ATS Although abnormalities. skeletal ) in slc2a10) ( gene ortholog SLC2A10 hybridization we observed that observed we hybridization situ in By slc2a10 that SLC2A10 gene, encoding for the is a maternal gene, since gene, maternal a is Diseases models maintains the maintains and slc2a10 Danio the same time, we generated a quadrouple transgenic fish overexpressing hAPP at overexpressing fish pathology transgenic tau quadrouple induce a to generated we and time, burden same Aβ the the increase further To HuC. promotor neuronal hTau or hTau its proteases, Presenilin1 or 2 (hPS1/hPS2) lead to an increase of Aβ of increase an to lead (hPS1/hPS2) 2 or Presenilin1 proteases, its the in found be can which mutations, (FAD) Disease Alzheimer’s Familial degeneration. neuronal and protein, tau microtubule-associated Aβ by characterized is and dementia protein of aggregates, intracellular form neurofibrillary tanglescommon and neuropilmost threads, whichthe consist ofis the (AD) Disease Alzheimer’s Butenandt-Institute, Adolf Ludwig-Maximilians-University, (DZNE), Munich, Erkrankungen Germany Neurodegenerative für Zentrum Deutsches F. van Bebber, A. Hruscha,M.Teucke, D.Paquet,C.Haass,B.Schmid Analysis of Alzheimer’s diseaseinducedneurotoxicityusingzebrafish Diseases models e hs te erfs a a aia mdl o vrxrs hg aons f hAPP orhTau of amounts high tooverexpress model an animal respectively to study aspects as of AD zebrafish the chose We aggregates isnotfullyunderstood. Aβ if and death cell Tauthe of role the Moreover species. neurotoxic the are aggregates or oligomers neuronal its and protein induces burden amyloid the how unclear still is It aggregation. high amountsof APP andTau proteininvivo. In summary, these transgenic zebrafish lines will serve as a valuable tool to monitor the effect of Progress intheidentificationof AβorTau inducedneurotoxicitywillbepresented. the transgenic fish to promote either aggregation of Aβ or to enhance aggregation of Tau protein. P301L P301L , and DsRed. To distinguish between toxicity induced by Aβ and Tau we will challenge incombinationwithDsRedasafluorescentmarkerproteinunderthecontrolof in vivo . We generated zebrafish stably overexpressing hAPP -Amyloid Precursor Protein (hAPP) or in one of one in or (hAPP) Protein Precursor β-Amyloid 308 42 , which is more prone to prone more is which , swe , hPS1 , 379 L166P swe ,

Posters Posters 380 will leadtoagreaterunderstandingofthecellulardysfunctionthatunderliesthisdisease. myopathy, centronuclear of study and the for model promising highly a offers muscle zebrafish in proteins associated and dynamin-2 of myopathy.analysis centronuclear The with associated mutations dynamin-2 express that zebrafish transgenic generating myopathy.currently Weare centronuclear with patients in seen phenotype the to leads function this cells, of muscle disruption that in and a network tubuloreticular plays the of dynamin-2 maintenance and formation that the in hypothesize role Wecritical tubules. (T-) transverse and reticulum the sarcoplasmic both of disorganization significant showed larval analysis individual ultrastructural Notably, fibers. of muscle hypotrophy and disorganization general revealed Immunohistochemistry embryos. At 48 hpf, morphant embryos show reduced response to mechanosensory stimulation. and larvae. At 24 hpf, spontaneous coiling is severely reduced or absent in dynamin-2 morphant larval stages. Behavioral analysis revealed early severe and embryonic motor during development defects muscle in skeletal on dynamin-2 focused analysis morphant subsequent edema Our embryos severe in resulted also It eyes. within thefirst48hoursoflife,despitepresencecardiaccontractions. and head underdeveloped and compartments Overall, muscle embryos. zebrafish developing on deficiency dynamin-2 dynamin-2 knockdown caused global of abnormalities, including a effect shortened body structure, the small determine dynamin-2 to of knockdown morpholino-mediated used we study, this In dynamin-2. human of development is unclear. In order to examine this question, we looked has been extensively studied in at intracellular processes, its specific functionthe in nerve and muscle zebrafish homologue centronuclear of form myopathy and a dominant form of hereditary motor and sensory neuropathy.dominant While a dynamin-2 disorders, neuromuscular human two cause to found were dynamin-2 in mutations Recently, where endosomes. newly-formed endocytosis, of release clathrin-mediated the to of contribute they components critical are dynamins Classical events. Dynamins are mechanochemical GTPases involved in a wide variety of cell and organelle fission University ofMichigan, Ann Arbor, USA E. Gibbs, A. S.Busta,J.Dowling,andE.L.Feldman Dynamin-2 FunctioninEmbryogenesis andSkeletalMuscleFormation 309 Diseases models f drnt slaae ucin Or eut poie motn isgt it te mechanisms the into insights important downregulation provide with resultsunderlyinglysosomalstoragedisorderpathogenesis. associated Our tightly function. is sulfatase signalling iduronate TGFβ of provide increased also that we assay, proof reporter molecular transactivation a a of of application the depauperation Through by precursors. cell mediated is patterning the cartilage that show Wealtered cartilages. craniofacial abnormal and trunk misshapen a including defects, developmental severe for early responsible be during may role downregulation its critical and a development withantisense vertebrate plays sulfatase function iduronate its that down demonstrate we Byknocking characterization oligos, morpholino and orthologue. identification sulfatase the iduronate of associated on zebrafish is report a broadspectrum of we glycosaminoglycans Here, sulfated a disorders. of metabolic with catabolism severe membrane-tethered with impaired congenital cell a byinteracting humans, and In functions matrix cellular extracellular morphogens andgrowthfactors. specific of exert chains proteoglycans glycosaminoglycan Sulfated Tübingen, Germany . Moro E. A novel functionalroleofiduronate-2-sulfataseinzebrafish earlydevelopment Diseases models 1 ae iess Uiest o Pdv, Italy; Padova, of University Diseases, Rare F. Argenton Department of Biology, University of Padova,Italy; of Biology,University of Department 1 R Tomanin R. , 1 2 A Friso A. , 2 A Mongera A. , 3 a Pac Isiue o Dvlpetl Biology, Developmental for Institute Planck Max 3 N Modena N. , 2 Department of Pediatrics and Center for Center and Pediatrics of Department epesn nua crest neural sox10-expressing 1 N Tiso N. , 310 1 M Scarpa M. , 2 and 381

Posters Posters 382 (1) HaftekZetal.,GeneExprPatterns.2003Jun;3(3):351-4. HSPG andFgfinthedevelopmentoforgans suchastheeye. interaction between its heparan sulfate chain and thus confirms the importancethe through ofpossibly pathway signalling aFgf control linkmay XVIII between collagen that suggests study Our was alsosignificantlyreduced. confirmeda study eye the of vascularization Finally, Ahistological the injected-larvae. hpf 72 in SU5402. cells retinal of FGFR inhibitor disorganization the to exposed larvae zebrafish in in disturbed in24 also lens was cells the ganglion retinal collagen around of XVIII MOs anarea differentiation injected-larvae. This The in phenotype morphants. was similar hpf to reduced 48 the retina specifically and phenotype hpf observed was signalling, Fgf8 of target a pea3, The lack of all 3 isoforms did not affect the expression of Fgf8. However, the transcription factor signalling. Fgf with link potential the and development eye the on MHB morpholino-knockdown XVIII collagen the as such development eye the for and the optic stalk. The aim of our study was thus to investigate the functional consequences of important structures some in XVIII collagen with colocalizes Fgf8 hpf, 24 At retina. the of differentiation the in role central a play to shown also was activity for binding proteoglycan) sulfate (heparan HSPG requires which signalling Fgf The for role a suggesting embryos the of collagen XVIIIinzebrafisheyedevelopment. size eye decreased in resulted variants three the against developing eye at 24 hpf and 72 hpf. In preliminary knockdown experiments, injection of MOs mRNAXVIII collagen express to found Wewere (1). the in expression transcript showed further ducts pronephric notochord, system, nervous central as such tissues Several variants. three the to hatching period using a probe complementary to the endostatin domain which is common to We previously reported on the expression pattern of collagen also inocularabnormalities. XVIII in zebrafish from segmentation results mice col18a1-null in XVIII type of lack The degeneration. vitreoretinal and with myopia high disorder recessive a syndrome, Knobloch cause XVIII collagen human of variant short in long) differing by their N-terminal domains are obtained from two distinct promotors. Mutations wide in occurs variety that of chains tissues side sulfate in heparan association with with collagen basement non-fibrillar membrane. a is Three XVIII isoforms Collagen (short, intermediate and IBCP, UMRCNRS5086, IFR128BiosciencesGerland, Lyon, France S. Bretaud,M.Malbouyres,F. RuggieroandD.LeGuellec Role ofCollagenXVIIIineye development 311 Diseases models 2 polb Diseases models 1 ee e ecie nvl N plmrs bt mtn (polb mutant beta polymerase DNA novel a describe we Here earlier and/or more affected during thedegenerationprocessifDNAare repairmachineryisnotfunctional. neurons some why explain therefore can and damage, DNA of levels higher induce turn, may,in properties intrinsic These explanation. one provide could more neurons these of of stress, oxidative for buffer production to capacity different and/or species to oxygen reactive leading activity metabolic of levels higher content, transmitter as such properties, intrinsic sensitivity,but increased this causing is what known not Todayis others. it than extent larger a to degeneration undergo and vulnerable more are neurons, of types some particular in machinery.types, DNArepair cell the Certain of components important in defects to linked been have diseases neurodegenerative and ageing premature cancer, of forms Some DNA polymerasesandDNA ligases. two are repair(NER) excision major nucleotide pathways. Key enzymes in repair these pathways and include DNA robust glycosylases, (BER) AP endonucleases, a repair with excision provided base are the which cells of of the machinery, activity groups base metabolic and normal sugar during of damages species repair To oxygen cell. reactive of production to due stress of the genome exogenous and cell death. The major endogenous and cause of damages to the DNAendogenous is oxidative to exposed constantly is events leading to DNAbody damages, which, if not repaired, can cause genetic the mutations, instability of cells all in DNA Genomic of Neurobiology Fessard,Alfred CNRS, Gif-sur-Yvette, France mechanism behindthisgroupofdiseases. this Thus, unchanged. can be used as a novel model for selective other neurodegeneration to elucidate some of the possible are for markers, patterning markers brain while it identities. whether neuronal exploring some currently of are earlyloss We specific a shows mutant reduced, as well is as markers populations, serotonergic neuronal and dopaminergic and week. of morphological one about abnormal Expression after die they show and response larvae escape weak Homozygous including phenotypes phenotype. behavioural null a generating thus triggers mutation this that show We frame-shift. a and 2 exon of out splicing- the to leading mutation point a carries mutant This screen. ENU hybridization-based C. Lillesaar Laboratory of Laboratory Development, Evolution and Plasticity of the System,Nervous UPR2197, Institute Department of Zebrafish Neurogenetics, Helmholtz Center Munich, Neuherberg, Germany and ne2385 –anovel zebrafish modelforneurodegeneration? 1 , G.Hausladen 1 , B.Hesl 1 , P. Vernier 2 andL.Bally-Cuif ne2385 1 312 mRNA degradation, mRNA polb ) identified in an in identified ) n situ in 383

Posters Posters 384 may underlieCdLSarepossible. are that while both complexes are subunits involved in cell division, alternative developmental suggest functions results that condensin These roles. non-proliferative of existence the supporting proliferation, of and with zones overlap completely not does Cohesin subunits cohesin of pattern cells. expression The complexes. two proliferating of distribution dynamically expressed in a similar, ofcondensin but not identical pattern, the indicating a diverse function of the withexpression it and compared subunits and embryo zebrafish developing the in cohesin subunits of expression the analyzed we cohesin, of roles non-canonical understand Tofurther hematopoiesis. and proteins maybeinvolvedintranscriptionalregulationofdevelopmentalgenes. patterning, skeletal cohesin that suggested been has pruning, it but understood, poorly far and so tissue-specific is underlying in mechanism The guidance roles key axon has as also but such division, cell development, in function only not does cohesin that recently shown been has It retardation. mental and anomalies, limb upper retardation, growth humans. CdLS patients have severe in developmental (CdLS) Syndrome abnormalities, Lange including de facial Cornelia dysmorphism, disorder genetic the cause to shown been have NIPBL, important have condensin subunits cohesin in and Mutations the respectively. condensation, cohesin chromosome and cohesion complexes chromatid sister multi-subunit in roles the division, cell During University ofOtago, Dunedin, New Zealand M. Moennich, S.Banks,andJ.Horsfield Analysis ofcohesinandcondensingenesduringzebrafish development 313 and SMC1A a wl a mttos n h chsn odn factor loading cohesin the in mutations as well as SMC3, Diseases models loss of Laminin function together with hypoglycosylation of alpha-Dystroglycan underlies the wide clinicalspectrumofsomeformsmusculardystrophies. underlies alpha-Dystroglycan of hypoglycosylation with together function Laminin of loss that and mechanism, glycosylation novel a through possibly Laminin, of secretion the on role processing. Taken together, our of results strongly suggest loss that Fukutin family to proteins play in a leads pivotal Dystroglycan of localisation Laminin absence of absence The function. Laminin of complete loss not but localisation, Dystrophin a Importantly, function. Laminin of loss with associated defects notochord elicits also but alpha-Dystroglycan of glycosylation reduced causes only not FKRP or Fukutin zebrafish of removal that show We (MOs). oligonucleotides allele and inhibited Fukutin family protein novel function a by recovered low-to-high have doses we zebrafish, of in antisense dystroglycanopathy morpholino Tomodel leading tothewiderangeofclinicalseveritystillremainsunclear. modification. The function of Fukutin and FKRP,sugar independent an stimulate may LARGE that however,suggesting patients, dystroglycanopathy some as well as the pathological mechanisms the from cells catalyze in defects glycosylation circumventsOver-expressing to LARGE alpha-Dystroglycan. of shown been have POMGnT1 and POMT2 POMT1, date, To brain with dystrophy muscular severe malformation of to onset a late congenital onset a of from milder limb-girdle ranging severity,muscular dystrophy clinical without of mental retardation. protein eerd o s ytolcnptis i wih uain afc a lat i kon r putative orincluding known six genes, least at affect glycosyltransferase mutations which in dystroglycanopathies, as to referred commonly are alpha-Dystroglycan of hypoglycosylation with associated dystrophies Muscular Kingdom Wellcome TrustInstitute,Sanger Wellcome TrustCampus,Genome Hinxton, Cambridge, United Y-Y. LinandD.L.Stemple dystroglycanopathy tolamininopathy from spectrum phenotypic activities the protein extends family zebrafish fukutin of Depletion Diseases models or fukutin (fkrp) and MO-injected embryos is mainly a consequence of defective post-translational defective of consequence a mainly is embryos MO-injected fkrp LARGE. Allelic mutations in each of these genes can lead to a wide spectrum POMT1, POMT2, POMGnT1, 314 fukutin, nonsense dystroglycan O-glycosylation fukutin-related 385

Posters Posters 386 fibers innervatedorsaltelencephalon,anareainvolvedincognition. histamine that show we as and those with accordance learning in are that affects results present system we memory.Here histaminergic the manipulating that shown have studies Previous stimuli. different to hypothalamus of response the modulate can systems neurotransmitter these histamine with together that suggesting hypothalamus, of functions physiological the mediating published in involved be previously to known also are with met-enkephalin and co-localized TRH GABA, accordance Galanin, rodents. from data in is is which histamine zebrafish in that TRH and show galanin was GABA, we with Nomet-enkephalin together, Taken neurons. met-enkephalin. histaminergic peptide, in observed opioid endogenous the containing fibers the and neurons histaminergic the between observed was pattern innervation Similar neurons. of the co-transmitters histamine in some neurons. TRH-immunoreactive fibers made identified close contacts with histaminergic we adult functions, zebrafish GABA, galanin and thyrotropin releasing hormone (TRH) were co-localized with physiological immunohistochemistry. by In zebrafish different of hypothalamus posterior the in of neurons histaminergic role these the and of in mechanisms underlying histamine the understand to order In rhythm. thirst circadianhunger, temperature, and body regulate to known is which hypothalamus the some in exclusively where telencephalon, found of are neurons part Histamine-producing the fibers. central ipsilateral the the with together telecephalic innervate to dorsal to side hypothalamus contralateral to anterior periventricular commissura the at of crossed fibers zone caudal the from with immunohistochemistry of larval zebrafish brains showed that histaminergic fibers projected combined mammalian microscopy confocal the behavior.3D anxiety-like to e.g. learning emotions, homologous processing amygdala spatialconsidered and is in pallium involved telecephalic selectively medial disorders The hippocampus, memory. and mammalian psychiatric the to homologous e.g. be proposed is to pallium in telencephalic lateral zebrafish, In by involved neurotransmitters. regulated other and functions histamine are neurotransmitter sleep and anxiety memory, modulatory Learning, diseases. neurodegenerative a is Histamine of Helsinki, Finland Neuroscience center and Institute of Biomedicine/Anatomy, Haartmaninkatu 8, 00014 University M. Sundvik andP. Panula andafferent co-transmitters innervation projections, telencephalic system: histaminergic zebrafish The 315 Diseases models 1 2 N. Malissovas A zebrafish modelof charcot-marie-tooth 2D Diseases models ubr f uat i dsrt sae o vle eeomn. n n o ti lns w named we lines, this of one In a development. weidentified valve metronome, of stages screen, discrete ENUmutagenesis in mutants scale of a large vertebrates number of In flow. life blood the throughout retrograde function prevent and to cells endocardial from derive valves Cardiac We will present our latest data on the mechanism of GARS function in this in vivo model of model vivo in this in function GARS CMT2D. of mechanism the on data latest our present will We during thedevelopingbrainoutliningventricles,besides aubiquitousbasalexpression. CMT.of that pathophysiology shown Moreover,the have in we insight an providing GARS, of dimerization proper the with interferes T130K that indications have we parallel, In hpf. 60 to up phenotype obvious no is there why explains which embryos, mutant (GARS) antibody we have shown that anti-human there is a maternally provided protein in the homozygous mutations an Using human NIH). NHGRI, severe Green, most Eric and the Antonellis Antony to with similar collaboration (in vitro, in cells neurons motor mouse in granules embryos. the phenocopied have We an as cellsignalling. bylinking synthesis ofprotein well as regulation translational step and transcriptional including tRNA), respective its to aminoacyl committed first functions the of catalyze repertoire (they broad translation a beyond have synthetases Aminoacyl-tRNA (L129P). mutation human atrophy type V. The mutation in our animal model (T130K) resides next to a previously described neuropathy,distal onset adult ToothCharcotmuscular Marie spinal and 2D type (CMT) disease glycyl t-RNA synthetase ). (gars Mutations in the human orthologue of that found have We 24. group linkage in siblings. We mapped the mutation with positional cloning and we located it in of clone myofibrils the DKEY-276I5 We present, hpf. is 96 actin by filamentous immotile although progressively that become found mutants these parallel, In development. valve cardiac impaired an of result a as onwards, (hpf) Developmental Biology, Biomedical Research Foundation, Academy of Athens, Greece Greece Athens, of Academy Foundation, Research aandBiophysics,Biochemistry University ofCalifornia, SanFrancisco, USA Biomedical Biology, Developmental gars T130K does not rescue GRS1 mutant yeast and is not associated with cytoplasmic homozygous 1 , D.Stainier mutants lack the organised fibrillar arrangement seen in wild type wild in seen arrangement fibrillar organised the lack mutants metronome mutants 2 andD.Beis metronome metronome show retrograde blood flow from 72 hours post fertilization post hours 72 from flow blood retrograde show 1 carries a recessive point mutation in the in mutation point recessive a carries metronome hntp b mrhln ijcin i wl type wild in injections morpholino by phenotype 316 gars are responsible for an is highly expressed highly is gars 387

Posters Posters Pediatrics, UCSF Helen Diller Family Comprehensive Cancer Center, San Francisco, USA. currently availablecancerdrugstotreattheprogressivephenotypes ofCFCinchildren. to propel forward the clinical discussion and foundation scientific strategy for assessing the suitability ofa using establishes and potential, therapeutic research and and activity medical allele for explore tool to geneticists tractable a as system zebrafish the shows work Our birth. after many of the phenotypic effects develop postnatally, patients may be helped by systemic unwanted developmental therapies effects of the drug. CFC syndrome has a progressive phenotype, and as a additional, the without embryo, the of with development early normal the restore can inhibitor MEK treatment which in window developmental a find we Importantly, development. early Treatment of CFC-zebrafish embryos with inhibitors of theof-function mutationsduringdevelopment. FGF-MAPK pathway can restore normal consistent with both the kinase-active and kinase-impaired BRAF CFC mutations acting as gain- during early development. BRAF CFC mutations promote an additive expressed effect when during development, outcome developmental equivalent the promote alleles mutant CFC impaired kinase- and kinase-activating both that find Wepathway. this of inhibitors chemical available and alleles disease human of function the assess to embryos zebrafish in alleles MEK and BRAF To study the developmental effects of CFC syndrome. CFC mutant of alleles treatment in effective therapeutically be hair might inhibitors such whether to as and skin question important the prompts pathway MAPK the of the inhibitors defects, active clinically of and availability phenotype, progressive a heart has syndrome CFC appearance, variants. kinase-impaired and activating facial dominant autosomal distinctive an kinase- a both promote BRAF in mutations syndrome CFC retardation. mental and abnormalities, syndrome, by (CFC) characterized cardio-facio-cutaneous syndrome develop the MEK2 in role or BRAF, central in mutations MEK1 a germ-line with plays born Children and cancers. most development, of progression human and formation for critical is pathway Ras/MAPK The Cancer Research, London,UK Biology,TransductionMolecular of Team,Signal and Institute Cell The for Centre UK Research 1 388 C. Anastasaki are and development zebrafish during sensitive tosmallmoleculeinhibitors active are alleles syndrome Cardio-facio-cutaneous Edinburgh, Western General Hospital, UK. Institute for Genetics & Molecular Medicine,Molecular & Genetics for Institute and Unit Genetics Human MRC UniversityThe of 317 1 , A. L.Estep 2 , R.Marais 3 , K. A. Rauen 2 University of California San Francisco, Department of 2 andE.Patton in vivo we have expressed a panel of 28 1* Diseases models 3 Cancer and affectedpatients. morphants in observed abnormalities eye the and inactivation Hccs between link pathogenetic these of role mechanisms in possible the pathogenesis a of MLS syndrome suggest and further data studies are this ongoing to explain Altogether the death. cell programmed and cell proliferation for important be to shown recently proliferation connexin43, and apoptosis. for Tunel expression and pHH3 of assays on morphants, domain revealed abnormalities abnormal in cell an RNAretina. neural ventral the of differentiation in defects and (RPE) epithelium retinal-pigmented the of formation abnormal an showed Syntaxin) Ath5, observed. also was microphthalmia, pigmentation blood displayed of morphants absence unexpectedly, addition, expected, In microcephaly. and As coloboma, condition. human the resembles which the protein (dominant-negative) and two mutated of the different both morpholinos of resulted injection in Instead expression phenotype. a aberrant pathological an phenotype, in result the not did studies function downregulate effectively morpholinos the that against the graded using directed lethality morpholinos Specific morpholinos. the embryonic mRNAor mutated the either possible of concentrations the (Oryzia overcome fish to medaka us in allow disease will this model for This model latipes). a generate to decided thus We cells. still stem out are knock- Hccs disease embryonic mouse this of lethality in early the anomalies demonstrated studies Previous developmental unknown. brain and eye the underlying mechanisms c and c1 necessary for proper functioning of the mitochondrial respiratory chain. The molecular apocytochrome both to heme of attachment the catalyzes that protein mitochondrial a codifies c-type linear skin defects, additional holocytochrome features include CNS malformation and mental retardation. the in mutations synthetase (HCCS) to transcript. Female patients display unilateral or bilateral microphthalmia and associated male- dominant disorder X-linked an neuro-developmental is lethal syndrome (MLS) defects skin linear with Microphthalmia The Genetics, of Pediatrics, Department Federico IIUniversity, Naples, Italy 1 A. Indrieri with linearskinlesions(MLS)syndrome Microphthalmia in observed phenotype the recapitulates medaka in Hccs of regulation Down Diseases models oeua Mdcn (EM, als Italy; Naples; (SEMM), Medicine Molecular eehn nttt o Gntc ad eiie TGM, als Italy; Naples, (TIGEM), Medicine and Genetics of Institute Telethon 1,2 HCCS transcript have been designed and injected. Our experiments have determined , I. Conte Analysis with specific markers (e.g, Pax6, Six3.2, Otx2, Rhodopsin, Crx, Chx10, Crx, Rhodopsin, Otx2, Six3.2, Pax6, (e.g, markers specific with Analysis 1 , G.Chesi 1 , P. Bovolenta 3 3 , B.Franco nttt Cjl CI, ard Spain; Madrid, CSIC, Cajal, Instituto 1,4 hybridization studies revealed studies hybridization situ in 318 ee Gi of Gain gene. olhccs 2 uoen col of School European 4 Medical HCCS 389

Posters Posters 390 that slc7a6osisrequiredforearlyneuraldevelopment. suggest embryos injected in observed the phenotypes of The knockdown. function gene morpholino-induced the investigated have We period. hatching the during line, lateral of neuromast and thymus the in also and embryogenesis, during system nervous central identified the putative of role biological the study to decided recently,we More of geneexpression. polymerase II subunit 3). This finding suggests that SLC7A6OS may play a role in the regulation Iwr1 Iwr1. probably significant level of sequence identity has been detected with or domains but low a polypeptide, this of role biological the about clues provide protein might that sites functional relevant identify to failed we Although vertebrates. in functions known of evolution, the encoded protein shows no significant sequence identities with other polypeptides soluble protein mainly localized a in is SLC7A6OS the that cytoplasm. demonstrated cells While HeLa Transientin the models. expression cell in pair gene We failed to obtain experimental evidence for a co-regulation of the RT-PCR studiesdemonstratedthatSLC7A6OS isexpressedinalltissuesandcelllineswetested. SLC7A6of (NAT) (SoLuteCarrierfamily7member A6) inthehumangenome. transcript antisense natural a representing gene coding protein novel a is SLC7A6OS ofBiomedical SciencesandBiotechnology,Department University ofBrescia, Italy A. Benini,L.Calvarini, A. Bozzato,F. Cignarella,S.Barlati,R.Bresciani,G.Borsani Functional characterization oftheSLC7A6OS geneindaniorerio mount that RT-PCRdemonstrated time studies cytoplasmic Real a cells. HeLa shows in expressed also transiently it when localization and counterpart human the to identical 46% teleost. is the protein of zebrafish genome the NATsin are genes two these that evidences no are there 319 is a maternal gene, expressed from the zygote stage to the adult fish. Whole fish. adult the to stage zygote the from expressed gene, maternal a is slc7a6os in situ is hybridization experiments showed that a factor that associates to associates that factor a slc7a6os zebrafish ortholog on chromosome 7 adjacent to RNA polymerase II through the binding to RBP3 (RNA RBP3 to binding the through II polymerase RNA slc7a6os is expressed predominantly in the SLC7A6OS SLC7A6OS gene is conserved during SLC7A6/SLC7A6OS cis-NAT Saccharomyces cerevisiae in Diseases models . We have We . rerio Danio gene by gene slc7a6os slc7a6, although Slc7a6os enable detailedstudiesonindividualgenesandnovelmechanisms. signalling, will which pathways, TGF-beta important mitochondrial several alters thus zebrafish in knockdown and PINK1 dysfunction. signalling stress, p53 and oxidative apoptosis pathway, signalling biosynthesis, factor inducible cholesterol hypoxia were program IPA different to available links have pathways commercially the on based these (p>0.04) altered significantly were of that pathways The PD. Most of forms migration. of cell specification amoeboidal and process, polarity biosynthetic axis glutathione morphogenesis, cerebellum development, are: specification of organ axis polarity, cerebellum formation, hindbrain formation, cerebellum pathways that were identified as significantly affected (cut off:the terms, GO their moreby pathway process biological organizedin were categories functional into than 50% of the genes altered) (level of confidence: p>0.01 and log fold controls change the values to from arraysfrom compared -1.6 morphants to PINK1 +0.9). When in the altered 237significantly genes being geneexpression genes 237 Wefound ontwo-colour and hybridised four Agilent. DatawasanalysedusingtheRandIPA from made programprograms. replicates were four fish of of set sets A different experiment. microarray a performed and oligonucleotides the understand to mechanism and order different pathways In that are involved. altered after PINK1 genes knockdown with and the morpholino pathways can of RNA information of unbiased amounts useful small give on Microarrays Pink1. for targets novel finding at aims study This and increasedsensitivitytocellularstressthroughadefectintheapoptosispathway. dysfunction mitochondrial to lead may mutations PINK1 death. cell apoptotic to threshold the lowers and inhibitors system proteasome ubiquitin to sensitivity enhances which of mutations mitochondria and cytoplasm. The function of the to localises product protein Pink1 The described. been have PD recessive autosomal cause by genetic alterations in some key genes causing hereditary PD. Mutations in the One of the challenges in Parkinson’s disease (PD) is identifying the biochemical pathways affected science center, CSC-ITCenter forScienceLtd, Espoo, Finland M. Priyadarshini Understanding PINK1mechanism by geneexpressionarrays Diseases models 1 Neuroscience Center, Institute of Biomedicine/Anatomy, University of Helsinki, Finland; Helsinki, of Biomedicine/Anatomy,University of Center,Institute Neuroscience 1 , J.Tuimala 2 , Y-C Chen 1 , P. Panula PINK1 1 is not clearly known but it is a protein kinase, 320 PINK1 gene that 391 2 Life

Posters Posters being investigated. is currently underway. Morphological effects due to the ablation of ablation the to due underway.effects currently Morphological is n ua HL cls eosrtd ht fish that demonstrated cells HeLa human in in vascular development. A more detailed phenotypic characterization of characterization phenotypic detailed Amore development. vascular in in iron metabolism. Morpholino-mediated knockdown of zebrafish of knockdown Morpholino-mediated metabolism. iron in role a play may counterpart, TRPML1 human the like gene, this that suggests (ICM), mass cell zebrafish genes during development. In particular, the expression of hybridization experiments show interesting peculiarities in terms of expression pattern of the two mount Whole compartments. endosomal/lysosomal late to localize mucolipin-1, human and of characterization functional the on polypeptides efforts our concentrated MCOLN MLIV,we on the focused of sequences the revealed using the presence of five different search fish genes related to human mucolipins. genome-wideBeing our interest and transcriptome- recently, we decided to study the biological role of the recently,of role biological the study to decided we for MLIV and subsequently we started to study the pathogenetic mechanism of the disease. More 1 392 A. Benini in mucolipidosistypeIV Identification and characterization of the putative co-orthologs of includes other two members: mucolipin-2 (TRPML2) and mucolipin-3 (TRPML3) encoded by encoded MCOLN2 (TRPML3) mucolipin-3 and (TRPML2) mucolipin-2 members: two other includes family mucolipin the storage mammals, In (TRP). channels cation potential receptor transient of lysosomal family recessive autosomal an mutations in the is 252650) MIM disorder that (MLIV,causes mental and motor retardation as well visual impairment. MLIV is caused by IV type Mucolipidosis of BiomolecularSciencesandBiotechnology, University ofMilan, Italy Department Department of Biomedical Sciences and Biotechnology, University of Brescia, Italy; 321 , the putative co-orthologs of human of co-orthologs putative the mcoln1.2, 1 , L.Calvarini and MCOLN3 MCOLN1 1 , S.Moleri genes, respectively. We have identified in the past the gene responsible gene, which codes for mucolipin-1 (TRPML1), a member of the large 2 , A. Bozzato 1 mcoln1.1 , S.Barlati MCOLN1Transient-expression. experiments MCOLN1 1 and , M.Beltrame mcoln1.2 gene in zebrafish. in gene zebrafish A MCOLN1, the gene mutated mcoln1.1 mcoln1.1 mcoln1.2 2 , G.Borsani Diseases models rtis smlry to similarly proteins, mcoln1.1 in the intermediate causes alterations causes function are also are function 2 Department Department morphants 1 mcoln1.1 in situ in lee. h mttos f h tre novel three the of mutations The alleles. novel of identification the to led that screen non-complementation a performed We Our laboratory identified the identified laboratory Our not reflectthehumancondition. Dystrophy,the Muscular of phenotype the though even for Muscular Dystrophy, one of the most common, lethal gene disorders. muscle Currently,structural the within the mutations mouse humans, null In mutant Australia rmtr so cdn cuig os f h gn pout To f hs mtns (dmd mutants these of Two product. gene the of loss causing codons stop premature broader variation of muscle fibre cross sectional areas. Due to its profound phenotype, the phenotype, itsprofound to Due and areas. sectional precursors dmd cross muscle fibre exhausting muscle of progressively interstitial variation by broader infiltrates, addition, characterised In are bymono-nucleated inflammation. mutants for homozygous indicative neutrophils, as replaced well as are tissue, connective myofibres Necrotic homozygotes. strategies fortreatmentofDuchenneMuscularDystrophy. the using thereby and rescue of the of severity the J. Berger A Fish ModelforDuchenne MuscularDystrophy Diseases models 1 os o la t a hntp rsu, n aohr n ta idcs tog kpig f exon of skipping the strong of induces rescue that a one to another leading and 32, rescue, phenotype a to lead that not 32 exon does of skipping mild a induces that one presented: are protocols skipping Twoexon us the unique opportunity to perform exon skipping experiments in live giving mRNAframe, mature reading the open from the exons disrupt these not of does either of repetitive rod domain, which is partially disposable for the function of Dmd. In addition, exclusion dmd of Medical Research, Melbourne, Australia; We documented extensive skeletal muscle wasting throughout the larvae lifespan of lifespan larvae the throughout wasting muscle skeletal extensive documented We humans. in observed that to comparable is atrophy muscle the of progression the that revealed orthologous the Australian Regenerative Medicine Institute, Melbourne, Australia; , the dmd, pc3 ta222a ) carry a null mutation in exons (32 and 34, respectively) that are located in the highly the in located are that respectively) 34, and (32 exons in mutation null a carry ) fishcloselyresemblessymptomsofthehumandisease. 1 , S.Berger mouse (Hoffman et al., Cell 1987), is the mainly used model for Duchenne for model used mainly the is 1987), Cell al., et (Hoffman mouse mdx gene (Bassett et al., Development 2003). Further survey of the phenotype the of survey Further 2003). Development al., et (Bassett gene dmd phenotype, we are in the unique position of comparing different levels different comparing of position unique the in are we phenotype, dmd 1 , A. Jacoby sapje ( 1 dmd , G.Lieschke zebrafish as a model to systematically test therapeutic tosystematically amodel as zebrafish dmd dmd pc2 homozygous phenotype. The data suggest that, due to due that, suggest data The phenotype. homozygous ta222a 3 lee (dmd alleles dmd Australian ResearchNeuromuscular Institute, Perth, ) zebrafish mutant as a carrier of a null mutation in mutation null a of carrier a as mutant zebrafish ) 2 , S.Wilton mouse is extremely mild and does and mild extremely is mouse mdx 3 , P. Currie Dystrophin ( pc1 , 2 dmd Walter and Eliza Hall Institute 1 322 ) lead to Duchenne to lead DMD) pc2 dmd zebrafish mutants. dmd and , pc3 null dmd encode ) dmd dmd pc2 and 393 ta222a ta222a

Posters Posters 394 aggregation statesandpotentialdegradationproducts. of the aggregates. re- Additionally, and we are dissolve performing biochemical astrocytes, and EM in analysis mobile to show are aggregate different during aggregates cytokinesis. Currently,process: dynamic we are a investigating effects is of various aggregates formation drugs on the that clearance of found We expression. GFAP of onset with detectable were aggregates positive established Rosenthal transgenic we zebrafish called lines mechanisms, expressing wildtype clearance and aggregates mutant possible GFAP intracellular and fused to dynamics eGFP of as aggregation a reporter. monitor component GFPfibers. To main the is GFAP AXD mutated In matter. white patients, of loss extensive causes astrocytes in defect primary a how unknown is for It gene disease. the the for causal are in protein, specific astrocyte mutations (GFAP), an protein acidic level, fibrillary glial molecular a rapid On by degeneration. characterized matter is white AXD disease. widespread genetic and human severe but rare very a (AXD), disease In a second approach, we use the zebrafish as model organism for the leukodystrophy Alexander to investigateeffectsonneuronsandgliaafterDTAin inductioninastrocytes. death body cell the of level of increased deformation an as deformed in zebrafish.well Currently, resulted as we arephenotypes, investigating astrocytes thetail” ablated ofan larvae“curly via of immunohistochemistry characteristic Ablation control and axis inducible. the heat is under turn (DTA) in system which cre/loxP A the via induced is protein subunit toxic the of toxin Expression promoter. specific astrocyte diphtheria expressing zebrafish transgenic points. time specific at astrocytes ablate genetically to aim we approach, first the In created We to theformationandmaintenanceofmyelin. astrocytes of contribution the investigate to strategies Wetwo we experiments. follow imaging live processes, extra-maternally and fast communication transparent, its neurons,to these developing larva, this Due species is particularly organism. suitable between to visualize illuminate model complex cellular interactions as in To communication zebrafish the with requires glia. work that non-myelinating process and regulated myelinating tightly a is Myelination Dept. ofBiochemistry, University ofMunich, Germany C. BrösamleandK.Stahl Glia-Neuronal Interactions andMyelination inZebrafish 323 Diseases models progression inall ALS patients. number large a opening avenues for the development of of therapies that could delay or prevent screening disease onset and the directly for thus TDP-43, mutant tested expressing zebrafish transgenic in be compounds pharmaceutical of then can lines These disorders. neuron motor model properly they whether determine to characterized functionally be will lines These lines. We are also generating of understanding specificbetter mechanisms of disease involved in mutant TDP-43 caused motor neurona degeneration. allow to as well as TDP-43 mutant us of allow partners molecular will unravel models to these vulnerability of selective characterization further the A TDP-43. in mutant involved to be neurons motor also of may function of loss specific a thus zebrafish; in phenotype similar a yielded TDP-43 of (AMO) oligonucleotide morpholino anti-sense using in toxicity neuron and motor motor neuron axonal selective defects phenotype in motor zebrafishspecific embryos. causes a Oncauses WT, themutant other of hand,Further,expression decreased not cultures. expression cord spinal but primary TDP-43, mutant of function of expression gain toxic since a through disorder neuron motor specific a causes TDP-43 mutant demonstrate that we Here embryos. zebrafish and neurons motor primary lines, cell in expression TDP-43 WT and mutant determine to use will I which N-terminus and C-terminus the both at ALS of patients. For cord this project, spinal we have the developed a a from series of bodies human TDP-43 inclusion cDNA of in constructs tagged aggregates that discovery protein the TDP-43, breakthrough encoding the made we Recently, (FALS). familial of number considerable are cases ALS of 10% Approximately movement. muscle control which cells neurons, motor of loss by characterized disorder neurological progressing rapidly adult-onset an is (ALS) sclerosis lateral Amyotrophic Université deMontréal, Canada ofPathologyDepartment andCell BiologyandCentre ofExcellence inNeuromics, E. Kabashi,L.Lin,P. Dion,N.Champagne,H.Durham,G.A.Rouleau,P. Drapeau Functional characterization ofmutant TDP-43 inzebrafish Diseases models Tardbp knock-outs in zebrafish as well as mutant TDP-43 transgenic fish TARDBPFALSgene in the mutations patients, (SALS) sporadic ALS and 324 395

Posters Posters 396 are morphants underlying CMT2A. MFN2 still is CMT2A in for model observed animal reliable mechanisms pathogenetic the dissect to tool useful and new a as zebrafish propose we lacking, abnormalities a that the Given pathology. human that the with consistent suggest results preliminary Our alterations arelikelyrelatedtothemovementdeficitsobservedinmorphantsatthisstage. these that suggesting larvae, morphant hpf 48 on observed were axons neuron motor abnormal an and eyes underdeveloped central nervous system the abnormalities and resembles optic and atrophy observed phenotype in some CMT2Athis Interestingly, patients. small Shorter ventricles. and brain the somites, of dilatation disorganized extensive tails, curved displayed they to antisense oligonucleotide usedamorpholino have knockdown and mitofusin function. we Morphant embryos showed motor CMT2A, impairment. Morphologically MFN2 ortholog of pathogenesis the zebrafish in the role identified its and gene this of function the investigate To by distalmuscleweaknessandatrophy. characterized neuropathy Charcot-Marie-Toothaxonal hereditary an 2A(CMT2A), type disease calcium of exchanges between modulation these the two cellular in compartments. MFN2 role gene a mutations maintain are play associated with may to it and essential cells is in mitochondria Mfn2 of organization reticulum. the endoplasmic the of surface the at and mitochondria of membrane outer the in located protein, GTPase dynamin-like large a is (MFN2) 2 Mitofusin ofBiology,Department University ofPadua, Italy G. Bergamin, A. Vettori, E.Moro,G.Polo,Vazza, N.Tiso, F. Argenton, M.L.Mostacciuolo Zebrafish mitofusin-2knockdown: anewmodelforCMT2Aneuropathy? 325 Diseases models two independent morpholinos independent two function nephrocystin-4 investigated we Here in thepronephrosforeffectiveurinaryexcretion. structure and motility are required in the Kuppfer’s vesicle to establish left-right asymmetry, and cytoplasmic protein Dishevelled (Dvl) is the central activator of both Wnt pathways. Proper cilia Wnt/PCP is required for polarized in convergent extension (CE) role movement during gastrulation. The pivotal a play cilia Primary regulating canonical Wnt/β-catenin cilium. and non canonical primary Wnt/PCP (planar cell the polarity) pathways. at localisation subcellular common a in mutations by Nephronophthisis, a heterogeneous group of autosomal recessive cystic kidney disease, is caused biology, UMR7622,Paris,France 1 ehoytn4 s clay rti rqie fr oto a wtbctnn ess wnt/pcp versus wnt/b-catenin 1 ao control for required balance andciliogenesisinzebrafish embryos protein ciliary a is Nephrocyatin-4 Diseases models interaction of interaction The CE phenotype suggests that abnormal CEduringgastrulation,left-rightasymmetrydefects,andpronephriccysts. nephrocystin-4 isrequiredforciliogenesisintheKuppfer’s vesicleandthepronephricduct. Moreover,Dvl. with interaction its via pathway,possibly Wnt/β-catenin the of repressor a and of Inmost was flow. Altogether our cells data fluid support a multiciliated role cilia-driven for nephocystin-4 as altered of both an activator an of motility the Wnt/PCP against nphp4-morphants, pronephriccystsarerelatedtoobstructionofthe cloaca. pathway cilia arguing However, in cysts, pronephric ciliogenesis. reduced in in preserved significantly nephrocystin-4 was of length role Cilia a motility. and length monociliated cilia cells of the Kuppfer’s vesicle and measuredpronephric duct of we defects, ciliogenesis to linked were formation cyst pronephric and defects laterality Tothe if investigate part ofthesameproteincomplex. β-catenin activation. We also demonstrated that nephrocystin-4, inversin (NPHP2) and Dvl2 are in HEK293 cells, we showed that nephrocystin-4 overexpression led to a significant decrease in the Using pathway. Wnt canonical the regulates nephrocysin-4 whether checked we Conversely, phenotype. CE the rescued Dvl2, myristilated-palmitoylated Hereditary nephropathies and kidney development, Inserm U547, Paris, France C. Burckly 1 H. Gaude with a core Wnt/PCP gene, Wnt/PCP core a with nphp4 , noig ehoytn. h nprcsis share nephrocystins The nephrocystins. encoding (NPHP1-11), genes NPHP 2 C. Vesque ® against nphp4 interacts with the Wnt/PCP pathway. We showed genetic 1 R. Salomon in zebrafish embryos. Both morpholinos Both embryos. zebrafish in nphp4 1 y os f ucin xeiet using experiments function of loss by vivo, in S. Saunier A specific activator of the Wnt/PCP, the of activator specific A vangl2. 2 S. Schneider-Maunoury -catenin reporter TOPFLASH reporter β-catenin nphp4-morphants, supporting 326 2 Developmental ® led to led 397

Posters Posters congenital muscular dystrophy, MDC1A. We have recently identified a zebrafish model of model a zebrafish identified recently MDC1A have called We MDC1A. dystrophy, muscular congenital (LAMA2) Laminin-alpha2 human the in Mutations Cardiology andBiophysics, Victor ChangCardiacResearch Institute, Sydney, Australia use a combination of pharmacological and genetic approaches to investigate the potential of potential the manipulating apoptosisforthetreatmentofcafdystrophicphenotype. investigate to approaches genetic and pharmacological of combination a use mortality.earlier with phenotype Lastly, in loss fibre since death, fibre the overall number muscle of affected fibresdelayed in this Despite apoptosis. undergoing before detachment after days several in that sarcolemma, which itself maintains its integrity. Using single fibre labelling techniques we show in retraction fibre contrast, Glycoprotein Complex (DGC), which leads to sarcolemmalin the rupture and subsequent necrosis. In model in zebrafish loss a Fibre detachment dystrophy. (sap), muscular fibre Duchenne`s sapje for dystrophin-deficient muscle characterized to previously the leading with model pathomechanisms the compared have We a cellularlevelcloselyresemblesthatofhumanMDC1A patients. Homozygous gene. 398 1 1 Modelling musculardystrophy andexploringtherapeutic strategies inzebrafish Australian Regenerative Medicine Institute, Monash University, Melbourne, Australia TE. Hall 327 mutants, the majority of retracted muscle cells remain viable but non-functional for non-functional but viable remain cells muscle retracted of majority the mutants, caf 2 IG. Huttner candyfloss ( caf 1 C. Sonntag mutants display a progressive muscle detachment phenotype, which on which phenotype, detachment muscle progressive a display mutants caf), which carries a loss-of-function mutation in the zebrafish is caused by attachment failure of the extracellular matrix to the to matrix extracellular the of failure attachment by caused is caf 1 PD. Currie caf is greater than in s u t dsuto o te Dystrophin the of disruption to due is sap ee eut n h ms cmo fr of form common most the in result gene is ultimately due to apoptosis, we apoptosis, to due ultimately is caf sap, leading to a more severe Diseases models 2 Molecular lama2 caf 1 A. Magnabosco The zebrafish modelforthestudy ofneurotoxinsmechanisms Diseases models h esml o tee prahd il ho nw ih o te oeua mcaim of mechanism molecular the on light information ontheprocessofregenerationneuromuscular junctionafterdegeneration. novel new provide will time same throw the at and neurotoxins will presynaptic diverse these by approached intoxication these of ensemble The at theneuromuscularjunctioncanbefollowed. resolved spatially and time a in mode. In addition behaviour fluorescent tagged toxinstheir can befollow used and theirto redistribution and localization and junction neuromuscular the of of process junction and ensuing this should allow the us to in incorporate fluorescent tags and/or on selected action protein components toxin neuromuscular the imaging of ofpossibility unique the offers animal this addition In regeneration. mode toxin the in of junction neuromuscular base the theof at components selected of role events the test to one allows which molecular tools genetic of range a offers andmodel cellular the of zebrafish The regeneration. junction neuromuscular of process the of and action of understandingmechanism the to aimed are In any case, these paralysis are followed by a complete recovery of function and our experiments and paralysisbyallowingamassiveentryofcalciumintothenerveterminal. PLA2 with The third group includes endowed spider neurotoxins, mainly latrotoxins, which cause are neurodegeneration toxins neuroexocytosis bycausingdegenerationofthenerveterminalandparalysis. presynaptic These family. target Elapid physiological their and activity the Asiatic and to Australian of belonging venom the from snakes isolated neurotoxins of consists group second The VAMP, SNAP-25 andsyntaxin(1). activity zinc-endopeptidase proteins: SNARE three the neuroexocytosis, of apparatus the of components key their the for specific following periods time long for junction neuromuscular the the genus of (TeNT);bacteria neurotoxin anaerobic tetanus by one produced and are (BoNT/A/G) they to The first group of neurotoxins includes the clostridial neurotoxins: seven botulinum neurotoxins on andtheirmodeofbinding,membraneinteractiontarget modification. information novel gather to zebrafish the of junction neuromuscular the on neurotoxins protein This project aims at studying the molecular mechanisms of action of three groups of presynaptic University ofPadua, Italy eatet f imdcl cecs Uiest o Pda Italy; Padua, of University Sciences, Biomedical of Department Clostridium, and cause botulism and tetanus, respectively. These neurotoxins paralyze 1 , M.Petron 1 , M.Rigoni 1 is the neuromuscular junction, where they block they where junction, neuromuscular the is vivo in , N.Tiso 2 , O.Rossetto 1 , F. Argenton 328 2 eatet f Biology, of Department 2 andC.Monteccucco 399 1

Posters Posters 400 knowledge ofpigmentvariationaswellthefunctionmelanosomesthemselves. our expanding greatly of capable system model a present we Takentogether melanosomes. to localize specifically to shown been has and genes melanophore-related for screen expression of gene global a morphology in identified was Melanoregulin a melanoregulin, of expressing protein melanosome line fusion inducible transgenic UAS a created have the we tool additional an analyze As genes. these of to products analysis micrograph wildtype, albinoandgoldenfishaswellimmunofluorescencetoshowthelocalizationof electron used have We understanding ofSNPsleadingtocolorvariationinzebrafish. better a achieve to hope we alleles multiple of analysis the Through exists. data functional little this, Beyond variation. color skin human in role a playing as described been recently also has same the of member a is It color.transmembrane family skin as SLC24A5, which human is affected in of the zebrafish mutant variation natural the with is associated and tightly 4, type albinism oculocutaneous cause to shown been MATPhas ortholog, human the while phenotype color coat underwhite the in implicated been has AIM1 mouse, in SLC45A2 of ortholog The (SLC45A2). 2 member 45, family carrier solute the in stop early an the containing isolated and mapped have We aszebrafish to studythebiologicalfunctionandtraffickingofmelanosomesinvivo . Also, thecell. system throughout model powerful a offer they cells, the dynamically in remain and visualized easily are them melanosomes traffic and bodies cell their within melanosomes retain melanophores zebrafish hair, and epidermis as example species for tissues, of other to evolution the in role melanosomes their substantial export melanocytes mammalian While disease. a in implicated being as well play genes these of Many kingdom. animal the across phenotypes pigment various with associated been have genes hundred a than More III, Max-Planck-Institute forDevelopmental Biology, Tuebingen, Germany C. Dooley,M.Konantz,H.Schwarz, C.Nüsslein-Volhard andR.Geisler human pigmentvariation zebrafish the of analysis functional and cloning Positional 329 uat o rgo lctd n hoooe 21 chromosome on located region a to mutant albino mutant, a novel model for model novel a mutant, albino Diseases models golden, and which identified in this screen to explore the hypothesis that bacterial virulence factors, bacterial factors, vivo essentialgenes,andhostgeneproductsareviabletargets forantibioticdevelopment. virulence bacterial that hypothesis the explore to screen this in identified compounds that rescue embryos from lethal currently are Wefor activity biological known infection. 1120with of compounds composed library attenuate chemical a screening that molecules small for screen of chemical feasibility a the conducting demonstrating challenge, lethal ciprofloxacin from antibiotics embryos anti-Pseudomonalinfected rescues also the imipenem have in or We embryos models. infected rodent of to immersion similar that infection, found combating for and required (lasR are sensing cells quorum myeloid and (pscD) secretion three type in involved that example, virulence factors required for infection are conserved from rodents to zebrafish (genes found have We countries. infection developed in infections nosocomial resistant, infection embryo zebrafish bacterium the a in screen chemical model. This will allow a us to identify compounds that rescue embryos from lethal challenge conducting with by properties antibiotic-like act (novel targets). We are exploring an alternative way of identifying small molecules that have could molecules antibiotic-like ways alternative as well as molecules antibiotic-like identify to and the growing demand for such drugs, there is a need to reconsider both novel methodologies traditional by antibiotics novel develop to ability our in gap current the non-essential but host the in viability for essential genes bacterial factors, virulence bacterial as such targets druggable potentially of range wide growth inhibit or kill that molecules small for bacteria Importantly,screening linezolid. and daptomycin drugs spectrum narrow the of development recent the of exception no novel classes of clinically relevant antibiotics have been discovered effective, in highly over been has 40 approach years, this with historically the While synthesis. protein and transcription, thegrowth growth, for inhibit essential or processes bacterial kill(bacteriocidal) to Historically, strains. ability bacteria resistant of their (bacteriostatic) antibiotic by of identified emergence been growing have antibiotics the combat to antibiotics of classes novel develop and identify to need clear a is There health. human to threat growing a is The increasing prevalence of antibiotic resistance among clinically important human pathogens General Hospital, Boston, USA 1 1 Identification of small molecules that modulate infection of infection that modulate zebrafish embryos molecules small of Identification Diseases models oeua Booy Ms. eea Hsia, otn USA Boston, Hospital, General Mass. Biology, Molecular AE. Clatworthy of zebrafish embryos resembles acute resembles embryos zebrafish of , one the one of the most common causes of antibiotic of causes common most the of one the one aeruginosa, Pseudomonas 1 J. Lee 1 EC. Hett Ti mtoooy eet fr ml mlcls ht target that molecules small for selects methodology This . vitro in 2 M. Mia 1 uh s el al ytei, N rpiain RNA replication, DNA synthesis, wall cell as such K. Mark P. aeruginosa challenge. We plan to use compounds neto i rdn hss For hosts. rodent in infection P. aeruginosa , and host proteins themselves. Given themselves. proteins host and , vitro in 2 S. Shaw 2 1 etr o Sses ilg, Mass. Biology, Systems for Center DT. Hung in aeruginosa pseudomonas 330 based methods based vitro in ) n zebrafish and mvfR)) ignores a ignores vitro in . aeruginosa P. 401 in

Posters Posters 1 J. A Lister Biochemistry,Bath,UK;of University 402 TemperleyN. Chemical-genetic screeningforsmallmoleculesthatalterpigmentcelldevelopment cell pathwayswillbediscussed. Combined death. cell for screening results melanocytes from this target screen will be specifically presented, and insight that into chemical molecules control of pigment small of suppressors as well as movement, and development irridophore and melanocyte for deficient are that lines type pigment cells, we are also screening for chemicals that suppress or enhance genetic mutant wild on effects the to addition In death. and environment, oftheir with interaction differentiation, andbiology migration, proliferation, their cells, ontogeny precursor/stem from development the their - affect cells pigment specifically the that compounds for libraries molecule small screening are we tools, molecular important identify to and lines, mutants genetic available the Tomelanoma. and graying, complement hair premature vitiligo, as such disorders, melanocyte to how these pathways may behave during human development and pigmentation, as well as in We aim to understand normal pigment cell development and regeneration pathways, iridophores. silver with the and xanthophore, yellow the a melanocytes, black/brown the crest: neural view the from derived are types cells pigment three the zebrafish, In differentiation. and patterning, Genetic screening in zebrafish has revealed a wealth of mutations in pigment cell development, Commonwealth University, Richmond, USA eiie UK; Medicine, MRC Human Genetics Unit & The University of Edinburgh, Institute for Genetics and Molecular 331 3 , R.N.Kelsh 1 S Colanesi S. , 2 ilg Porme o Rgnrtv Mdcn, eatet f ilg and Biology of Department Medicine, Regenerative for Programme Biology 2 , andE.Patton 2 P Rengtved-Lundegaard P. , 3 Department of Human and Molecular Genetics, Molecular Virginia and Human of Department 1 1 K Taylor K. , 1 H Ishizaki H. , Diseases models 1 I J Jackson J. I. , 1 , subsequent and morpholino by knockdown rescue onthedevelopmentofzebrafishembryos. OCRL of effects the studying organism, model a as zebrafish utilising syndrome Lowe to leads OCRL of loss the how elucidate to aim we Here cause thediseasephenotype. activity,and expression protein of loss to lead which mutations, these how unknown currently is it However Lowe). of syndrome (oculocerebrorenal OCRL1 termed enzyme, 5-phosphatase causing births, polyphosphate inositol male expressed ubiquitously a within mutations 100,000 to mapped been has disease in 1 affects which disease disruption X-linked to specific rare tissues, namely a those of is the brain, eyes syndrome and kidney.Lowe The genetic cause of the Cellular Systems, University ofManchester, Manchester, UK M. Hughes,Lowe,andI.Barinaga-RementeriaRamirez Zebrafish asamodelorganismforlowe syndrome Diseases models 332 403

Posters Posters Canada. of Society Parkinson the of chapter Ottawa the and CIHR by Funded humans. in PD of etiology the studying that hoped is It dpf. 3 at densities lower in neurons DA mis-patterned mildly show embryos surviving while zebrafish both of knockdown double Remarkably, diencephalon. ventral the in neurons DA of patterning perturbed and neurodegeneration mild databasewhich 2inthe of Knockdown (MOs). oligonucleotides morpholino antisense blocking exon translation designed we located, codon start the themis-annotated With sequencing. and PCR of by confirmed was ORF upstream a putative identified We The tissues. fetal and adult human combined, these expression patterns are comparable to the ubiquitous expression of adult brain and liver. No liver. and brain adult tissues. heart and liver muscle, brain, adult the in expressed is and dpf 7 to (dpf) post-fertilization day 1 designated parl1 and 404 rhomboid the include proteases. which of family (I-Clips) proteases intramembrane-cleaving called proteases, regulated of in role (PARL) a plays place in the rhomboid-like hydrophobic environment of it the lipid bilayer. Thus, it is executed by where a unique class presenilin-associated membrane mitochondrial conserved intramembrane inner proteolysis (RIP). Surprisingly, the evolutionarily this typically hydrophilic enzymatic in reaction takes the located protease, for codes death and PINK1 including genes, of number a in mutations with associated is (PD) Parkinson’sdisease Familial Biology,University ofOttawa,of Canada Department (CAREG), Genomics Environmental in Research Advanced for Center S. NobleandM. Ekker Role oftheparlgenesinParkinson’s DiseaseetiologyusingDaniorerio asamodel same pathway as pathway same 333 parl2 . Another candidate gene that may play a role in dopaminergic (DA) neuron (DA) dopaminergic in role a play may that gene candidate Another Parkin. maternal and PINK1 , the rhomboid-7, parl2. We found that expression is low, but is highly expressed from 1 - 7 dpf, as well as in the transcript was detected in the heart and muscle tissues. When tissues. muscle and heart the in detected was transcript parl2 genes in zebrafish will help elucidate their contribution to the to contribution their elucidate help will zebrafish in genes parl . Zebrafish have two have Zebrafish Parkin. start codon is not annotated by the Ensembl database. Ensembl the by annotated not is codon start parl1 orthologue, is genetically upstream and in the in and upstream genetically is orthologue, Drosophila parl1 is not expressed maternally, but is present from genes results in 52% - 87% lethality, 87% - 52% in results genes parl paralogues, which we arbitrarily we which paralogues, parl or parl1 Diseases models results in results parl2 parl seen in rnfrain A io sre uig pnl f nw MP ad I-iae niios was inhibitors PI3-kinase and MAPK known of carried outtofurthervalidatethismodelasapotentialdrugscreeningvehicle. panel a using screen pilot A transformation. promising model to screen small molecule libraries for entities that could antagonize neoplastic melanomas. The early onset and easy visualization of the mitfa-V12HRas phenotype makes this a activation of MAPK and PI3-kinase signalling, both pathways are often over activated in human increased exhibited also Melanophores stages. larval early very from dysplasia and hyperplasia melanophore marked exhibits model tumour The therapeutics. anti-cancer novel for screening zebrafish mitfa promoter, results in tumour formation in transgenic fish the within using 4 weeks, melanophores enabling zebrafish in V12HRas oncogenic Expressing group. our in established been recently has models neoplasia relevant a such intriguingly screens, throughput high such the issues associated with target-based discovery.of Zebrafishmany is a vertebrate model uniquely suited to circumvents that discovery,strategy alternative Phenotype-guided discovery.an provides drug screens, genetic cancer chemical are are using of techniques rate encountered success Newcomplementary problems the increase main attrition. to stage required The late malignancies. and of validation identification, treatment target for candidates clinical of numbers adequate delivering not is discovery drug target-based that apparent increasingly is It Molecular CancerStudies, University ofManchester, UK P.A. Walker, ME.Jones,and AF. Hurlstone screening Developing an early onset melanoma model in zebrafish suitable for forward chemical genetic Cancer 334 405

Posters Posters f esr Cl Booy Ogngnss Cnr d Rglcò eòia Breoa Spain; Barcelona, Genòmica, Regulaciò de Centre Organogenesis, & Biology Cell Sensory of $ *Hubrecht Institute – KNAW and University Medical Center, Utrecht, the Netherlands; 406 S. Choorapoikayil*, A. Faucherre The RoleofPTENinDevelopment andCancer cell growth, proliferation and survival. Functional analysis in mouse, in analysis Functional survival. and proliferation growth, cell pathway,PI3kinase/Akt/PKB the counteracts PTEN that known is it general PTEN is one of the most frequently mutated tumor suppressor genes in progression of cancer. In well asinvivo. tumorigenesis during PTEN of role the investigate we cells tumor of transplantation and culture cell zf like techniques art the of state Using progression. cancer studying for PTEN after survival lacking zebrafish of cell tool powerful the use to Now,enhanced focus siblings. we to compared γ-irradiation and proliferation cell enhanced exhibit mutants Double defects. defects. Up to day five the embryos show severe malfunction marked as hyperplastic-dysplastic through early embryogenesis, however viable and fertile. Concomitant are embryos mutant and changes loss phenotypic induce to appear not leads does function of loss Single to a dramatic phenotype. Double knock out model mutants genes, pass these in since development, during two encodes lethality. genome role embryonic zebrafish causes The deletion PTEN organisms, important an plays PTEN that demonstrated Institute ofBiologyLeiden, Leiden University, theNetherlands 335 ptena and ptenb, which seem to have a partially redundant role during early development. #, P. vanDuijn*andJ.denHertog* from 3 dpf onwards the embryos display developmental $ and Drosophila which is involved in involved is which # Laboratory Laboratory as vitro in C.elegans Cancer pten at the level of the anterior aortic bifurcation, as shown by loss of expression in this region of region this in expression markers of endothelial loss the by shown as bifurcation, aortic anterior the of level the at impairment of the systemic circulation in zebrafish embryos. This is paralleled by an interruption of down Knock the expressioninbrainislimitedtodiencephalonandmidbrain. spinal cord. At 48 hpf of expression the hpf 24 At precursors. system nervous central in and somites within expressed strongly is cxcr7 RT- by investigated fertilization (hpf) onwards] and by whole mount in situ hybridisation. During late somitogenesis was of development levels significant during [with expression analysis its PCR and cloned was ortholog cxcr7 zebrafish the Here, angiogenesis. tumor in receptor this for role a suggesting thus vasculature, Recent chemokines. CXCL12/SDF-1 and ITAC CXCL11/tumor-associatedby normal expressed the is by CXCR7 not that but shown vessels have studies binds that receptor chemokine a as and identified been has CCX-CKR2) Sciences (RDC1, CXCR7 dissemination. metastatic and growth tumor in Biomedical role important an plays ones, pre-existing of from vessels blood new of growth the Angiogenesis, Department Immunology, and Pathology General Biotechnology, University ofBrescia, Italy of Unit G. DeSena,S.Buraschi,C.Tobia, andM.Presta Cxcr7 andtumorangiogenesisinzebrafish Cancer embryo. Also, ourdatasupportaroleforthischemokinereceptorintumorangiogenesis. In thatcxcr7 conclusion, found we ourlaboratory, pro-angiogenic mammaliantumorcells. in with grafted are embryos zebrafish when angiogenesis developed tumor inhibits strongly assay downregulation xenograft zebrafish/tumor a using with cxcr7 morpholino showed a delay in the development of intersomitic vessels. Furthermore, cxcr7 appears to be involved in the development of the vascular system in zebrafish expression by injection of an ATG-targetingof injection cxcr7by severe expression a to leads morpholino in the brain is maintained, associated with a distinct expression in the in expression distinct a with associated maintained, is brain the in cxcr7 cxcr7 is expressed in branchial arches and in posterior lateral line whereas and fli1a :EGFP) embryos injected embryos tg(kdr:EGFP) transgenic Also, VE-cadherin. cxcr7 rncit en dtcal from detectable being transcript 336 hus post- hours 3 407

Posters Posters downregulation of the of downregulation observed Notably,we response. apoptotic the in genes prominent of expression differential in f h cl cce akr popoyae-itn-3 Additionally, phosphorylated-Histone-H3. marker, levels cycle persistent cell and caspase-3, the activated of marker, apoptotic conserved the of levels reduced displayed embryos transgenic Irradiated radiation. ionizing to exposure following cycling cell in vivo transgenic zebrafish harbouring the human The its of therapies. targeted of design the to zebrafish is a robust model for lead studying vertebrate hematopoiesis and leukemogenesis, by and virtue leukemogenesis of mechanisms universal unprecedented opportunity for chemical modifier screens to identify promising therapeutic promising to identify data screens these agents thatrestoreanormalphenotype. modifier chemical phenomenon, for anti-apoptotic opportunity unprecedented the underlying with mechanism a together suggest Taken differentiation. myeloid of upregulation of zebrafish Preferential fate. myeloid predominantly a to precursors cell NUP98-HOXA9blood reprograms phases ‘definitive’ and ‘primitive’ both affect hematopoiesis, including the recently-described erythro-myeloid progenitors (EMPs), suggesting to appeared transcription gene in Kai ppoi gns Tee aa otlt sprsin f aps--eedn aotss s a as apoptosis caspase-3-dependent of suppression postulate mechanism for data These genes. apoptotic to Health Centre, Halifax, Nova Scotia, Canada,Scotia,Centre,NovaHalifax, Health eatet of Departments 408 Berman J.N. hematopoiesis intransgenic zebrafish Oncogenic etbae eaooei. lcdtn te ciiy f oncogenic of activity the Elucidating hematopoiesis. vertebrate highly-conserved the to in prognosis (CML). leukemia myeloid chronic in crisis blast as well as (AML), leukemia inferior with associated is and The Cancer Institute, Boston, U.S.A. lysC, genes, myeloid-specific of expression upregulated to leading hematopoiesis, embryonic radiosensitivity.restore to required (2) be may interruptions bcl-x 1,3 337 NUP98-HOXA9 fusion oncogene results from a t(7;11)(p15;p15) chromosomal translocation , A. T. Look and ex utero utero ex phenomena: (1) L have suggested insufficiency of single-target therapy, and that synergistic, multi-target synergistic, that therapy, and single-target of insufficiency suggested have , and downregulation of the erythroid-specific gene, erythroid-specific the of downregulation and l-plastin, 1,2,3 upess ellr ppoi ad ergas myeloid reprograms and apoptosis cellular suppresses NUP98-HOXA9 development, and conserved genetics and cell biology. We have engineered a engineered have Webiology. cell and genetics conserved and development, HOXA9-mediated oncogenesis , A. M. Forrester M. A. , during ‘primitive’during pu.1 terminal of impairment an implies further hematopoiesis 4 1 irbooy n Imnlg and Immunology and Microbiology pro-apoptotic gene, and upregulation of the of upregulation and gene, pro-apoptotic puma NUP98-HOXA9 leads to suppression of cellular apoptosis and abnormal HOX gene family of developmental transcription factors is critical for critical is factors transcription developmental of family gene 1,3 , E. R. Boyd R. E. , mdae lueoeei ad rvd an provide and leukemogenesis NUP98-HOXA9-mediated NUP98-HOXA9 4 in vivo . ‘Rescue’ experiments using a morpholino Department of Pediatric Oncology,Pediatric Dana-Farberof Department 1,3 n tetetrltd ct myeloid acute treatment-related and novo de , C. Grabher C. , 2 eitis Dlose University, Dalhousie Pediatrics, translocation, and report two novel perturbs zebrafish NUP98-HOXA9perturbs 4 , S. Da’as S. , NUP98-HOXA9 NUP98-HOXA9 3 Tee changes These . gata1 , J. T. Dobson T. J. , and bcl2 HOXA9,belonging a reveal may bcl-x Cancer resulted 1,3 L , F-B. ,

pu.1, 3 anti- IWK 1 C-D Heidecke 2 I.J. Marques Metastatic behaviour ofprimaryhumantumoursinazebrafish xenotransplantation model Cancer n ocuin or eut so ta te erfs i is mroi sae isauseful specimen. stage embryonic its in zebrafish animal model for rapid analysis of the invasion and metastatic behaviour of primary human tumour that show results our conclusion, In a functionalvasculature. cells showed no metastatic behaviour when injected into cloche into injected when behaviour metastatic no showed cells and metastatic behaviour, invasiveness whereas demonstrated primary embryos, control zebrafish cells remained 72hpf in of the liver liver.the in Pancreatic tumour implanted organotopically was which absent transplantation, when non-tumour after tissue hours was implanted. 24 Furthermore, primary within human tumour formation cells, micrometastasis when and invasiveness Xenografts of primary human tumours implanted into the yolk of real-time invasion,migrationandmicrometastasisformation. 48hpf zebrafish embryos showed dye excellent an are live embryos zebrafish transparency, its fluorescent to Due embryos. with zebrafish 48hpf tumours into transplantation after human behaviour metastatic their gastrointestinal investigated and (CM-Dil) from explants small labelled We model toanalysetumourinvasionandmetastaticbehaviour. For primary human tumours we establish here a simple, fast, sensitive and cost-effective in vivo living in metastasis and invasion tumour organisms of to cancer,date Analysis is difficult including formation. and involves diseases, complex metastasis and time many and consuming investigative of techniques. invasion progression cell drives migration cell of the regulation Aberrant of Institute Cardiology Interuniversity & Utrecht Centre Netherlands, Utrecht Medical University & Institute Universitätsklinikum Greifswald, Klinik für Innere Medizin A, Greifswald, Germany.Greifswald,A,Medizin Innere für Klinik Greifswald,Universitätsklinikum Netherlands.Zoology,Leiden, IntegrativeThe of Biology, of University of Department Institute 1 , F.U. Weiss 2 , M.Lerch 2 , D. H. Vlecken 2 and C.P. Bagowski 1 , C. Nitsche 1 2 , J. Bakkers 3 , A.K. Lagendijk -/- mutant embryos, which lack which embryos, mutant 338 model to follow to model vivo in 3 , L.I. Partecke 3 Hubrecht Hubrecht n vivo in 409 2 ,

Posters Posters 410 capacity andmetastaticbehaviourofthishighly-aggressivepancreaticcancercellline. migratory the for important are RIL and LMO7 Mystique, genes PDZ/LIM three the that suggest siRNAs led to reduced migration and inhibition of the formation of micrometastasis. These these of one either with cells cancer results pancreatic of treatment embryos, control to comparison In or LMO7(PCD1;FBX20). RIL (PDLIM4) SLIM), (PDLIM2, Mystique for (siRNAs) RNAs interfering small gene-specific with post day microscopy. PaTu8988-Tscanning embryos, zebrafish into 2injections to Prior treated were cells of laser confocal sac by behaviour metastatic yolk and invasion monitored the and embryos into (dpf) (PaTu8988-T)fertilization cells cancer progression pancreatic and human formation implanted We and metastasis follow migration to on able genes were xenotransplantation PDZ/LIM zebrafish we a of model, Using down cells. cancer knock pancreatic human of of effects behavior metastatic the analyzed we study, this In can they trait, important associate withtheactincytoskeleton. an share family PDZ/LIM the of proteins all Functionally, (LMO7). 7 Enigma Homolog, and ZASP), the two LIM kinases (LIMK1 and LIMK2), and the LIM only protein (Enigma, subfamily Enigma the of the three (ALP, RIL) of subfamily and ALP Mystique, genes Elfin, ALP-like, five domains: LIM several or one and domain, PDZ a both for encode that genes domains. additional assorted and functions biological diverse with proteins largeof a variety in present modules, interaction are domains LIM as well as PDZ Institute ofBiology, ofIntegrative Department Zoology, University ofLeiden, The Netherlands A. I.Belo,J.Marques andC.P. Bagowski in metastatic behaviour ofhumanpancreaticcancercells RIL and Mystique LMO7, genes PDZ/LIM the for role a reveals model engraft zebrafish A 339 In zebrafish, we identified eleven identified we zebrafish, In n ra time. real and vivo in Cancer 24(9):427-30. Rambaldi, D and identify to validate thenoveltumorsuppressorgenes. used strategy the as well as discussed and presented be will obtained data the the most significantly repressed candidates in our melanoma model. Allof expression thisthe workrestore to isbe will ongoingstep next and The lines. cell melanoma in and tissue melanoma whereas melanomas Weoncogenes). (potential up-regulated are few a only human in analysis similar a perform will zebrafish cancer in candidate genes) novel tumor-suppressor (potential many down-regulated of is expression genes the that demonstrate skin) normal (versus tissue in genes 101 the of expression of several melanoma samples. Data obtained by Real time-PCR analysis analysis from zebrafish melanoma quantitative out carried we model melanoma our in re-express to suppressor tumor candidate the of choice the towards step first a As networks. protein in connectivity high the and duplication of frequency low the for selected were novel genes 101 test specifically will we genes candidates suppressor known tumor Besides candidate cancer genes identified system. with a of bioinformatics approach UAS (Rambaldi et al., expression 2008). the These using the cells restore melanoma in to approach this use will We of geneexpression. in the same cells that undergo transformation, thus allowing precise tumor-specific manipulation activity oncogene of modulators of down targetexpression/knock for allows system binary This line resultsinmelanomadevelopmentwithahighfrequency. 1 V. Anelli Validating novel cancergenesinazebrafish melanomamodel Cancer UAS system, where the crossing between a melanocyte-specific Gal4 line with a UAS:H-RAS Gal4- the on based is that zebrafish in progression melanoma of model a established Wehave IFOM, The Firc Institute of MolecularBiology, 1 , C.Santoriello et al, (2008) Low duplicability and network fragility of cancer genes. 1 , D.Rambaldi 2 , F. Ciccarelli 2 IEO, Oncology, InstituteofEuropean Milan, Italy 2 , M.Mione 1 340 Trends Genet 411 V12

Posters Posters 412 H. Ishizaki Melanocytes forCellDeath Chemical-genetic ApproachesZebrafishin and Novel Identify Yeast Sensitize that Compounds 1 Medicine, Crewe Road South, Edinburgh, UK;Edinburgh,South, Road CreweMedicine, the melanocyte. for vulnerability of point new a reveal may mechanism BIO1E7 the that suggest we apoptosis, in melanocytes mouse on situ. Given BIO1E7 that melanocytes are have of inherent mechanisms to effects coop with DNAthe damage induced and action, BIO1E7 of mechanism direct the to BIO1E7, indicating that the BIO1E7 mode-of-action is conserved. We are currently exploring find that mouse melanocytes deficient for the nucleotide excision repair are especially sensitive we yeast, in Like apoptosis. and arrest, cycle cell G2/M-phase a (ROS), species oxygen reactive the DNA damage repair pathways. In human deletion melanoma gene cancer with genetically interact to BIO1E7 identify to us enabled has sensitivity compound for mutants yeast cell 6000 lines, we of findprofiling BIO1E7genetic induces One Systematic pathway. zebrafish. tyrosinase-independent by in biology melanocyte of aspects death cell for melanocytes zebrafish sensitize specifically distinct to appears BIO1E7 compound, novel alters that panel molecule small a and lead compound discovery. Screening 1600 bioactive compounds has allowed us to identify used in the drug-discovery process for target validation, disease modeling, toxicology and target widely becoming are zebrafish screening, Wellchemical and genetic high-throughput to suited live tissueaswelltheirprogressiontomelanoma. in melanocytes of visualization the allows which system, model zebrafish the using processes death - may be highly informative for new therapeutic approaches to melanoma. We study these stem cells, their proliferation, migration, differentiation, interaction with their environment, and Understanding the ontogeny and biology of the melanocyte - their development from precursor/ agents. cytotoxic to resistant naturally are melanocytes capabilities, anti-apoptotic and survival highly motile neural crest cells, melanoma has melanoma high is why so to difficultmetastatic year. Clues to one treatpotential, than may less come from of and the expectancy nature life armed of median the a melanocytewith having itself. individuals Derivedafflicted enhanced from with chemotherapy,all to resistant and cancers, aggressive most the of one is melanoma Metastatic Road,Edinburgh, Mayfield UK Building, Swann Michael Edinburgh, of University The Sciences, Biological of MRC Human Genetics Unit & The University of Edinburgh, Institute for Genetics and Molecular 341 1 , M.Spitizer 2 , J.Widenhain 2 , D.WMelton 2 Wellcome Trust Centre for Cell Biology,Cell WellcomeSchool for TrustCentre 1 , I.J.Jackson 1 , M.Tyers 2 , E.Patton Cancer 1 1 J. Holzschuh Functional analysisofthetumourigenictransactivator Cited1inzebrafish Cancer a subset of restricted to the subventricular zone of the hindbrain. We found, that zfcited1 is co-expressed in spatial restricted pattern in the neuroectoderm. After somitogenesis zfcited1 expression becomes a in 18hpf at at and hybridization vesicle situ otic in the by in detected 11hpf first is expression mammalian the of ortholog great parts cancer formation. To better understand Cited1 functions we have cloned the zebrafish state. Developmental processes like proliferation and differentiation of progenitor cells mirrors in few functional data on Cited1, which suggest that Cited1 might keep cells in an undifferentiated have hCITED1 also been miss-expression. shown its to be through reliable their markers malignancies of papillary other regulate thyroid and carcinoma the (PTC). melanoma and Yet,in of implicated there progression p300 are been only a previously and has CBP hCITED1 Human integrators responses. transcriptional transcriptional dependent the bind that domain activator (Cited) domain carboxyl-terminal ( D)-rich acid family. All (E)/aspartic family members have in acid common the highly glutamic conserved carboxy terminal transcription with transactivator The Non-DNA binding transactivator Cited1 is the founding member of the Bonn, InstituteofHumanGenetics, Bonn, Germany the zfCited1canhelptounderstandhCITED1functionintumor formationandprogression. of analysis functional further that reason we b-catenin bind to zfCited1 and hCITED1 of ability involved in regulation proliferation and to hold the progenitor status of cells. Despite the to different bind b-catenin in a LEF/TC reporter luciferase assay. In summary we conclude that zfcited1 is able not is zfCited1 that show we hCITED1 the to Contrary b-catenin. binding by pathway Wnt fast a show can we construct degradation zfCited1-GFP of the a zfCited1-GFP protein after using mRNA Indeed injection. hCITED1 negatively same D-box. regulates the the a contains additionally zfCited1 and of O-box sequence protein putative The O-box. an through hCITED1 of not is degradation proteolytic phenotype reported the the by neurogenesis. explained cases be could disturbed both phenomenon This In and penetrant. fully mRNA. hindbrain hCITED1 theusing observed of is size phenotype same in The mRNA increase zfcited1 dramatic and a morpholino show p53 and a survive with co-injected Embryos dependent apoptosis. p53 dependent in p53 resulted zfcited1 of down apoptosis knock presumably as an off gain target morpholino effect. The over and expression of Azfcited1 mRNA function also induced experiments. of loss function performed of we zfcited1 of function the elucidate To differentiation. mutant notch-pathway expressing neurons differentiated freshly or nvriy f riug Booy , eeomna Booy Febr, Germany; Freiburg, Biology, Developmental 1, Biology Freiburg, of University pcna expressing proliferating cells, but never in neuron precursors expressing 1 and A. Zink , where too many neuronal progenitors undergo neuronal undergo progenitors neuronal many too where bomb, mind 1§

and analyze its role in development. Zebrafish zfcited1 Zebrafish development. in role its analyze and Cited1 is lost in the in lost is zfcited1 of Further,expression elavC. 342 CBP/p300-interacting § nvriy of University deltaA 413

Posters Posters Commonwealth University, Richmond, USA; tissues ofyoungzebrafishaswellarangetumourcores. cores in a tissue microarray. Successfully validated antibodies will be tested on both developing tumour medaka and zebrafish human, both using optimisation to approach systematic highly a taking are We peptides. human of regions conserved highly lackofreliable against raised antibodies of the panel this. However, for and a optimising are we this to nevi response model In effort. this hamper may anideal field with the in antibodies zebrafish be individuals of to zebrafish numbers the large allows generating melanoma of ease The progression. melanoma during changes molecular and cellular the study to characteristicstools effective and important are Antibodies behavioural and cellular histological, examine between melanomasarisingfromdifferinggeneticbackgrounds. to us allow will models these and Am Hubland, Würzburg, Germany; Würzburg, Hubland, Am 414 J. Richardson Molecular andGenetic Approaches toMelanomaDevelopment 1 MAPK pathway by pathway in MAPK mutation activating constitutively the is nevi of mutation initiating frequent most The nevi. benign from is occur can melanoma which in way One rate. The incidence of melanoma is rapidly rising, and treatment of advanced stages has a low success acr eerh ete UK; Centre, Research Cancer rngnc ad ae f td o mlncts n tmu dvlpet Cosn zebrafish Crossing development. tumour and melanocytes the of carrying study of ease and and genetics tractable transgenics their of because melanoma study to zebrafish utilise We activated. are The Institute of Genetics and Molecular Medicine, MRC Human Genetics Unit & Edinburgh Edinburgh & Unit Genetics Human MRC Medicine, Molecular and Genetics of Institute The 343 ptenb, we are able to generate melanomas arising in differing genetic backgrounds. Utilising BRAF 1 , J.Bartlett V600E BRAF mutation with lines containing loss of function mutations in mutations function of loss containing lines with mutation 1 , I.J.Jackson V600E causes proliferation of melanocytes before senescence pathways senescence before melanocytes of proliferation causes 2 hsooice hme , nvriä Wrbr, Biozentrum, Würzburg, Universität I, Chemie Physiologische 3 Department of Human and Molecular Genetics, Virginia Virginia Genetics, Molecular and Human of Department 1 , M.Schartl 4 Hubrecht Institute,Hubrecht Utrecht, The Netherlands 2 , J. A. Lister (V600E). Activation of the of Activation (V600E). BRAF 3 , J.denHertog 4 , E.Patton p53, Cancer ptena 1 source of Sdf1 in vitro. To additionally investigate guidance of tumor progression in vivo, we vivo, in established progression transgenic tumor medaka of lines which guidance specificallyinvestigate express Toadditionally a vitro. to in attracted Sdf1 are of chemokine, source the not but receptors expressing Sdf1 express Xmrk which that cells, melan-a indicate mouse assays migration Trans-well melanoma. invasive by colonized Xmrk and Sdf1 in the same cell will contribute to a better understanding of the influence of influence the of understanding better a to contribute chemokine signalsonmelanomaprogressionandtumorspreading. will cell same the in Sdf1 and Xmrk overexpressing lines transgenic double of investigation Further vivo. in movement cell pigment transient experiments show that individual cells expressing increased levels of ye ivsiae. xetoa hg epeso of expression high Exceptional investigated. types RT-PCR analysis revealed different expressional levels of xanthoerythrophores into diverse pigment forms of in pigmentary lesions, expressed including melanoma. exclusively Quantitative of is levels receptor,High cells. factor Cxcr7 growth epidermal and the of Cxcr4 version mutant receptors corresponding active constitutively a Xmrk, oncogene a itsthe model this in In model. melanoma medaka genes transgenic orthologous and the characterize to started Sdf1/Cxcl12 we detail, in of progression melanoma during role the investigate To metastasizing tumorwithverylowsurvivalrates. the of aggressive, levels highly a melanoma, High malignant human organs. in identified been distinct have Cxcr4 Sdf1-receptor in nodules tumor secondary of formation and progression tumor drive to suspected is it as interest, great of become has Sdf1/Cxcl12-Cxcr4 pair recpetor tumor cells exploit chemokine signals for progression and metastatic guidance. The chemokine- the in identified first lymphatic system molecules, due to their role signal in the migration and secreted homeostatsis of immune cells. small However, of family abundant an are Chemokines Wuerzburg, Germany Hubland, Biozentrum,Am Wuerzburg, of University I, Chemistry Physiological of Department D. Liedtke,I.Erhard,B.Wilde, J.Laisney, S.Meierjohann,M.Schartl Role ofSdf1inmelanomaprogressionandpigmentcellmovement inmedaka Cancer expression result in malignant transformation of melanophores and melanophores of transformation malignant in result expression xmrk and sdf1b sdf1, cxcr4 and sdf1 a dtce i tissues in detected was cxcr4b genes in pigment cells. Initial 344 cxcr7 in different tumor sdf1 show altered 415

Posters Posters 416 activity, i.e.progenitor/stemcells. to generate a transgenic zebrafish that will allow us to identify have characterized the minimum regulatory sequence of the zebrafish telomerase gene in order Finally, we fins. regenerating in upregulated was expression telomerase that fish. observed Wealso senescent in declined drastically length telomere and life. activity zefrafish telomerase the both However,during increased length telomere and expression telomerase that found and for about telomerase activity and telomere study length among a broad manageable range of tissues complete and ages from zebrafish a perform we and Thus, cells. stem of fecundity behaviour the in withmammals length high telomere and size, similarity small andphysiological including advantages, highthroughput screening. ofgenetic We several have used this offers model to investigate degree the it roles played by ahigh telomerase and shows zebrafish The Little is known about the expression and functions of telomerase in non-mammalian vertebrates. telomerase and length telomere activity haverecentlybeenproposedasstemcellmarkers. Therefore, activity. telomerase lack tissues and cells somatic However,mouse. and human of cells tumor and tissues most adult several of compartment cell stem the in expressed is and telomeres of maintenance the in role primary Telomerasea plays Arrixaca”, Murcia, la de “Virgen instemcell Hospital telomere Spain University Surgery, of and Department Unit, Research role of telomerase the M. study Anchelin, L.Murcia, F. Alcaraz-Pérez, E.García, M.Moreno,L.Cayuela to model behaviour a new zebrafish, The 345 in vivo cells with high telomerase Cancer normal cellcycleandmaybecomeausefulvesseltoscreendrugstargeting them. during regulators mitotic of role the study to tool valuable a provide could model zebrafish the human PICH ortholog, which is critical for the sister-chromatida separation. Our indeed results also suggest that is zPICH the that demonstrate data These morphants. the for cytokinesis and separation chromosome incomplete found and microscopy lapse time performed we detail, in division cell the analyze to order In segregation. chromosome proper in zPICH of role crucial When suggesting embryos. observed, frequently were bridges anaphase zebrafish knocked-down, was zPICH of level in the proliferation vigorous undergoing regions cellular the at highest was expression zPICH PICH. human to similar metaphase, to prometaphase from kinetochores to localized and Plk1 with associated readily was zPICH bioinformatics. though (zPICH) PICH for ortholog zebrafish identified we First, cells. cell mitotic of abundance and manipulation genetic normal of during easiness the PICH for model embryogenesis of zebrafish adopted importance we Therefore and lacking. are division function the exploring studies PICH, in interest checkpoint also involved (Plk1-interaction in DNA decatanation during sister PICH chromatid separation. Despite the considerable partner interaction its helicase). PICH and kinases is a putative component of spindle assembly checkpoint mitotic Plk1 as a tension sensor and the in Among interested development. became drug we anti-cancer for targets attractive became and disease of genetic instability, cancer. Mitotic kinases orchestrateeach the to spindle sister-chromatidsassembly checkpoint of daughter cell. Failure in this process separation results in the loss of genetic even information and may cause the ensures checkpoint assembly spindle mitosis, In University, National Seoul Sciences, Natural of College Republic ofKorea Sciences, Biological of Department K-H. Jeong, J-Y. Jeong,H-O.Lee,andH.Lee Zebrafish PICHplays acriticalroleinsister chromatid separation duringmitosis Cancer 346 417

Posters Posters p53 silencing/up-regulationontheexpressionoftrim8genes. 1 A zebrafish genome-wide search allowed us to detect two orthologs, named animal model. this in role biological and developmental its clarify to order in phenotype, function of loss-of and expression analyze to is goal Our through mechanism p53. suppression with tumor interaction a the studies in and involved be astrocytomas, could and it suggest glioblastomas cells of mammalian in number a in down-regulated is gene TRIM8 cancer.brain human in the rearrangements that and revealed deletions investigations in Recent involved mostly region a 10q24.3, the within located is and brain in expressed highly is TRIM8 activation ofp21inmammaliancelllines. that four p53responsiveelementslocatedinthefirstintronofTRIM8gene.Importantly, weshowed Bionformatic analysis, ChIP and luciferase assays revealed that genetic diseases,cancerdevelopmentandviralinfection. of biological processes and are implicated in several pathological conditions such as Mendelian structure, which suggests a common basic function, TRIM proteins are involved in a broad range named motifs by characterized proteins the encode of TRIM family.presence the gene TRIM/RBCC the of member a is TRIM8 II, Paolo San Giovanni Rotondo, Giovanni Italy Poliambulatorio Sofferenza, della Sollievo Casa IRCCS Laboratory, Genetic trim8.1 and of TRIM8 in brain tumors onset through p53 interaction, we plan to test the reciprocal effects of involvement the on light shed to and cells mammalian in obtained data the confirm to order In trim8 genes. zebrafish both and each of silencing the of consequences the evaluate to order in morpholino, Zebrafish motives. and located on chromosome 13 and 12 respectively. Bioinformatic analysis indicates that both 418 M. Manzoni Molecular andfunctionalcharacterization oftrim8genesinzebrafish and retina. We are now generating central nervous system (forebrain and hindbrain) and eye, in particular in the cerebellum, tectum stages of development. et Boeia Sine ad itcnlg, nvriy f rsi, Italy; Brescia, of University Biotechnology, and Sciences Biomedical Dept. 347 TRIM8 interacts with and increases the p53 protein stability and it affects p53 transcriptional trim8.2 possess the p53 responsive elements and encode a protein with the typical tripartite trim8.2 knockdown/over-expression on 1 , L.Micale -box and an associated an and B-box and trim8.1 tripartite In situ hybridizations show a similar expression pattern in the developing 2 , G.Paganini otif, which consists of a of consists which motif, xrsin s eetd y TPR trig rm early from starting RT-PCR by detected is expression trim8.2 trim8.1 and 1 , E.Monti oil region (RBCC). In spite of highly conserved highly of spite In (RBCC). region Coiled-coil trim8.2 knockdown by microinjection of specific 1 , G.Borsani p53 expression and, vice versa, the effect of ING finger, one or two zinc-binding two or one finger, RING 1 andG.Merla TRIM8 regulation is modulated by Danio rerio TRIM8 rerio Danio 2 trim8.1 and ortholog(s) Cancer 2 Medical trim8.2, trim8.1 transposable elements from the maize, we first introduced zebrafish K-ras oncogenic of expression the control to system recombination Cre/loxP and system expression gene LexPR mifepristone-inducible the of combination a used Wenext research. long-term for Sciences Laboratory, Singapore; 1 reporter gene under the liver-specific the under gene reporter EGFP novd n uoieei ae lo eeuae i or K-ras our in deregulated also are tumorigenesis in involved pathways Other up-regulated. significantly are pathway signaling MAPK the in involved genes A. T. Nguyen inducible cre/loxsystem Generation and analysis of zebrafish liver cancer by overexpressing k-rasv12 using mifepristone- Cancer tumor formation in this organ and thus demonstrated the general application of the inducible the Cre-loxP systemindevelopmentoftransgenictumormodelszebrafish. of application general the demonstrated thus and organ this in formation tumor to transgenic zebrafish expressing K-ras expressing zebrafish transgenic to K-ras of expression profiles gene the analyze to employed then tumor was of approach possibility microarray cDNA the A raising metastasis. fish, aged some in tumors GFP-positive some noticed also we and adult and fry both from induced be could tumor liver the that found we system, transgenic inducible the using By examination. histological our in depicted as carcinoma, hepatocellular may help to investigate molecular mechanisms and search for signature pathways leading to leading pathways signature K-ras for search and mechanisms molecular investigate to help data may microarray of analyses Further oncogene. to kras of known expression miRNAs of regulation several in of involved be down-regulation the indicated data array miRNA our this, with of K-ras oncogenesis, we developed a zebrafish liver cancer model through generation of generation through model cancer K-ras liver activated expressing zebrafish lines a transgenic stable developed we oncogenesis, K-ras of mechanisms molecular the Tostudy tumorigenesis. in drive K-ras that mechanisms molecular elusive. remains cancer human Although in and Ras transduction of signal understanding of complete edge a leading yet oncology,the molecular at that of been has signaling to Ras resembling system model cancers. promising closely human a study as tumors emerged recently zebrafish has ) rerio (Danio of zebrafish the studies human, the expression gene and histology With in EGFP-positive F EGFP-positive in transgenic zebrafish. We observed tumor growth in EGFP-positive livers but also a high mortality Department of Biological Science, National University of Singapore, Singapore; Singapore, of University National Science, Biological of Department V12 V12 induced liver oncogenesis. We recently also applied the mifepristone-inducible system mifepristone-inducible the applied Wealso oncogenesis. recently liver induced mostly phenotypes, tumor liver induced successfully system this of Activation . KRAS mutations occur in vast majority of human cancers, very little is known about the 1 , A.Emelyanov V12 induced liver tumor formation. Several Ras oncogene family members and members family oncogene Ras Several formation. tumor liver induced 1 zebrafish, which hindered the maintenance of these transgenic zebrafish transgenic these of maintenance the hindered which zebrafish, 2 , C.H.V. Koh 3 Department ofMicrobiology,Department University, State Oregon USA V12 in the pancreas and have also successfully induced successfully also have and pancreas the in 1 , J.Spitsbergen lfabp V12 promoter and obtained germline transmitted germline obtained and promoter n h lvr Using liver. the in 3 , L.Sun V12 rngnc oe. Consistent model. transgenic 1 348 kras , S.Parinov V12 Activator/Dissociation oncogene fused with 2 andZ.Gong 2 TemasekLife 419 1

Posters Posters 1 and G.Basso 420 3 2 E. Rampazzo cells derived glioblastoma human xenotransplanted for niche reliable a represents brain Zebrafish tumours andCSCsbehaviorstudies. mimicking an phenotype. immature GBM the preserves microenvironment CNS all transplanted human As cells. Hif-1α. Our preliminary marker observations hypoxic indicate that the at 48 and hpf stage, Nestin used zebrafish we controls, CD133, of expression the for checked were cells and analyses immunocytochemical By dye. fluorescent a successfully into the brain and could be tracked, fertilization. Transplanted cells were followed for at least 10 days post injection. Cells integrated post- hours 48 at embryos, albino zebrafish of ventricles xenotransplanting brain into cells for GBM-derived primary parameters optimized We niche. cell tumour reliable and In cancer.transparent a novel cure a to as exploited aimed been tools has embryo and zebrafish the strategies studies, designing recent in help can cells, cancer and affecting preserving microenvironment, hypoxic been the has characterizing parameters GMB the in of CSCs understanding of presence The GBM. deeper a Thus, niches. tumours in brain of typical microenvironment hypoxic the with relapses associated of incidence high the to related probably most are and Nestin) and BMI1 MSI1, SOX2, CD133, (i.e. markers primitive express cells stem of subsets These (CSCs). cells stem cancer as known properties, stem-like with in cells occurring tumours, malignant primitive of subsets important common functionally contain to shown been has which system, nervous central most the the of one is (GBM) Glioblastoma eaoOclg Lbrtr, eatet f eitis Uiest o Pdv, Italy; Italy; Padova, Padova, of of University Biology, University of Pediatrics, Department ofNeurosurgery,Department University ofPadova, Italy Laboratory, of Biology Department Developmental Laboratory, Hemato-Oncology 349 1 1 , N. Tiso in vivo antisense riboprobes and anti-human nuclei antibodies, able to detect to able antibodies, nuclei anti-human and riboprobes ACTBantisense hypoxic microenvironment, appear as valuable biosensors for human brain 2 ,

F. Pistollato 1 , G. Del Moro in vivo , by GFP expression or vital staining with 3 , A. Della Puppa in situ in hybridisation, tumour grafted tumour hybridisation, 3 Thus, zebrafish embryos, by embryos, zebrafish Thus, , G. te Kronnie model to recreate to model vivo in 1 ,

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Women’s Hospital and Women’s Hospital Cancer Institute, University of Utah, Salt Lake City, USA; Departments Departments of S. L.Perkins J. K. FrazerK. J. Heritable Zebrafish TCellCancerModelsfromaMutagenesisScreen Cancer important classofhumancancers. of models new identified have several we with sum, T-LBL In samples, far.and T-ALL. control thus These mutants should provide exciting experimental seen platforms matched for study of this aberrations not number but copy tissues, recurrent malignant in losses or withcopy gains regions number neoplastic have revealed to analyses contributing genomic lesions comparative genetic Specifically, non-inherited transformation. additional of each discovery in the risk cancer and inherited line conferring mutations the cloning on focused are efforts Current mutants three all Overall, correlates. human recapitulate humanTcellmalignancy, showingheritablediseasepredisposition. their like cells, leukemia-initiating contain and their clonality.verify cells transplantable, radiation-sensitive, relapse-prone, cells are Malignant T-ALL. Expression profiles confirm neoplasms while are T lineage,mutant, and each T cell inheritance, receptorIn analyses human resemble morphologies cellular and infiltration, histologic incidence, ofdisease develop. T-LBL tumor and dominantto malignancies other with for exhibit that required are demonstrating ‘hits’ mutants incomplete, genetic mutants is somatically-acquired different penetrance Twodisease three, ofthree predisposition. all In recessively. acts cancer one characterization cell and T identification heritable the describe we Here, phenotypes, includingthedevelopmentofGFP cell T abnormal for Then, stages juvenile at (lck::EGFP). progeny their expression screened microscopy, we GFP fluorescence via cell-specific T with zebrafish transgenic adult the mutagenize of randomly germline to ENU used We neoplasia. cell T of models genetic new create to diseases these in lesions genetic other Tounknown. are deficiency,this designed address screen phenotypic many zebrafish a pioneered we However, cases. T-ALL of 50% about pediatric isincomplete. in occurs common pathogenesis molecular their are activation NOTCH1 as established, is malignancy cell T in mechanism transformation key One into (T-ALL) insight Consequently, finding. leukemia cytogenetic pathognomonic no carry neoplasias and cell T most cancers, childhood (T-LBL)many Unlike morbidities. lymphoma significant cause often treatments their and prognoses, poor lymphoblastic have diseases These malignancies. cell T USA 1,2 6,7 , N. Meeker N. , , andN.S.Trede 1 Pediatrics, 5 Harvard Medical School, Boston, USA;Boston,School, Medical Harvard 1,3 , L. Rudner L. , 2 Oncological Sciences, 1,2

2 , D. F.D. , Bradley + tumors. 3 Internal Medicine, and 1,2 , A. C. H. Smith H. , C. A. 4 Department Department of Pathology, Brigham and 7 ARUP Institute, Salt Lake City,Lake Salt Institute, ARUP 350 1,2 , K. Brown K. , 6 Pathology, Huntsman 4,5 , C. Lee C. , 421 4,5 ,

Posters Posters 422 specificity ofsmallmoleculechemicalinhibitorsdesignedtoothertarget proteins. evaluate rapidly to model vivo in efficient an as used Our be can zebrafish that them. suggest also studies among PLK1 to specific most was PLK1-Yang and activities off-target different had of phenotypes the to zebrafish that are genetically deficient of plk1. These data suggest that the three PLK1 inhibitors similar were which structure, develop head deformed and properly axis body shorter didn’t had ON01910 PLK1-Yang with with treated Embryos normal. relatively was treated structure head the while tail and Embryos trunk normal. relatively were LFM-A13 embryos initiated, is program added with treated Embryos transcription compound. each for phenotypes different showed but When mRNAdays 3 for survived zygotic cell. when large stage one later into a fuse stage, shield to at cells caused and dividing from embryos prevented inhibitors these of all stage, 2-cell at added inhibitor,PLK1-Yang.When PLK1 homemade one and ON01910, and LFM-A13 inhibitors, PLK1 published two selected specificity, we evaluate to used be human. can and zebrafish zebrafish if between determine structure To 3-D identical which share very high homology with their human notthoroughly counterparts. In particular, PLK1 has a is nearly this issue. inhibitors Bioinformatics analysis revealed zebrafish genome has all of the PLK family members these of specificity examined. Here, the we use whole zebrafish embryo assay However, coupled with genetic analysis to address candidates. drug anti-cancer of PLK1 small molecule inhibitors have been developed as chemical biology tools and potential therapy.cancer targetfor good a as regarded been Ahas it number tumors, several in regulated chromosome segregation, and cytokinesis in cell cycle. Since PLK1 expression is abnormally up PLK1 (Polo like kinase 1) is one of the key regulators which control mitotic entry, spindle assembly, Graduate SchoolofPeking University,Biotechnology,Shenzhen Shenzhen,and China Biology Chemical of School Genomics, Chemical of Laboratory S. Xin,J.Lu, Y. Zhao,N.Wang, S.Li,C.Z. Yang, H.ZhongandS.Lin Evaluation ofSmall-moleculePLK1InhibitorsUsingZebrafish 351 Cancer to conditionally express mouse express conditionally to the using by model zebrafish transgenic a established system have Tet-on we formation, cancer liver in involved processes biological underlying its as well as to c-myc of function the investigate order In undetermined. remains genes target its and progression and initiation tumor the in transcriptomic analyses and we expect that these experiments will provide new insights into insights new provide will mechanism ofc-mycinvolvinginthelivercancinogenesis. experiments these from that pathway expect c-myc we the and of analyses understanding transcriptomic for out carried be will studies Microarray growth. and development of stages different during formation cancer liver in role its investigate to ages transgenic activate to going are we experiments, ongoing the In reversal. and the potential of our indicated observations Our blood. including tissues other to migrated and detached were cells liver that embryos treated the of some in observed also we addition, In stages. developmental from of 21 dpf, activation less time the is needed to to obtain liver Compared overgrowth when phenotype. liver enlarged the alleviated and expression was overexpressed for a when longer time. adenoma Brief withdrawal hepatocellular of Doxycycline of greatly Histological development reduced the the by followed phenotype. and stage early such at liver showed siblings mouse of non-transgenic overexpression that the showed analysis of (n=23) abdomen, enlarged apparently none showed fish whereas transgenic of 74) (n= Doxycyclin 100% inducer dpf, the 21 with from treatment starting of days 14 After promoter. ) protein binding acid fatty have indicated that in transgenic zebrafish studies has only been proved in function its zebrafish, APC-deficient an T-cellin adenomas leukemia.hepatic and adenomas Despiteintestinal in thatfound these studies rodents. of models Although transgenic in shown been also has C-myc ( University, State Oregon USA Z. Li in system transgenic zebrafishtet-on a in c-Myc of expression inducible by tumor liver of Formation Cancer 1 Department Department of Biological Science, National University of Singapore; 1 , H.Zhan MYC ) is frequently observed to be overexpressed in human cancers. Its role in tumorigenesis 1 , Z.Zeng c-Myc transgenic zebrafish as a model in the study of liver cancer progression c-myc is involved in tumorigenesis, the exact mechanism of 1 , J.Spitsbergen in the liver under the control of the zebrafish the of control the under liver the in c-Myc 2 , andZ.Gong in zebrafish liver caused hyperplasia of hyperplasia caused liver zebrafish in c-Myc 1 c-mycbeen has overexpression c-Myc was activated from early 2 Department Department of Microbiology, 352 c-Mycdifferent at c-myc involving lfabp ( c-Myc c-Myc c-Myc 423 liver

Posters Posters 424 system totesttheefficacyofmitoticinhibitors,letaloneassessfunctionskinases. powerful model in studying mitosis. Furthermore, zebrafish embryogenesis is an efficient formation and that its loss primarily results in mitotic arrest. Our study reveals that zebrafish is a spindle bipolar the for essential is zAurA that demonstrate Takendata together,death. our cell and/or mitoticslippageinmostcells.Intermittently, wealsoobservedchromosomebridgesand delay mitotic displayed morphants the live-imaging, the microscopy.In time-lapse out carried of fate the analyze to and zebrafish order H2B-GFPtransgenic Histone generated we zAurA, blocking after cells In individual morphants. the to similar spindles, multipolar and monopolar arrest. The embryos treated with MLN8054 also exhibited abnormal spindle formation including manifested monopolar spindles and increased expression morphants the of in mitotic cells the markers as progression suggestive mitotic abnormal of the mitotic to attributable be might defects zebrafish developed short trunktreatment, withinhibitor the severeor injection growthmorpholino the retardationAfter inhibitor,MLN8054. andmolecule small cell a death. or morpholino The blocking developmental translation a utilized we (zAurA) Aurora-A in zebrafish of block function experiments to the order loss-of-function In development. and performed cycle cell we normal stages model Here, the poor. as embryos, zebrafish is organisms and cells normal in Despite the emerging interest for clinical applications, our understanding how Aurora-Aas promisinganti-cancerdrugs. functions and induction of cell death with its inhibition, Aurora-A specific inhibitors drew much attention centrosome and spindle function. Since the discovery of Aurora-A overexpression in cancer of control cells the in roles implicating mitosis, during microtubules and centrosomes at localized is Aurora-A is a serine/threonin kianse, which regulates many intricate processes during mitosis. It University, National Seoul Sciences, Natural of College Korea Sciences, Biological of Department H-Y. Jeon andH.Lee Zebrafish isapowerful systeminstudying Aurora-A functioninmitosisanddevelopment 353 Cancer in vivo ouain f el. n both In cells. of population reporter and if so whether cells expressing GFP were found in the existing tubules or in a distant do that not express kidneys either reporter.pronephric The goal differentiated was to determine terminally if we could in visualize reactivation of damage either for assayed We development. methodologies thatcouldeventuallybeusedasanalternativetherapyforacutekidneyinjury. develop to hope we numbers, their manipulate to able being and cells reactivated the of origin the determining by future, the In cells. progenitor of population local a to ascribed be possibly Our current data suggests that the GFP positive cells are localized outside the kidney and could for damage in the larval zebrafish kidney.tubular proximal mediated Togentamicin modelinduced we damagedevelopment inembryonic real-time,and werepair utilizedkidney transgenic lines regeneration. kidney of study the for between model link suitable the and explore simplified To respond to renal injury by nephron neogenesis. The zebrafish pronephric kidney thus provides a the for to able therapies are and ability new regenerative remarkable a injury.of possess kidney Fish acute of development treatment the for clues important provide may repair nephron of intra-renal versus extra-renal cells is poorly understood. A source better of proliferating cells that repopulate the understanding injured nephron, and the relative contribution of of the mechanisms parallels with the growth and maturation that occur during kidney organogenesis. However, the and actively proliferate differentiate to to known reconstitute tubular are epithelium. cells The tubular regeneration process epithelial is renal thought to phase have recovery many the During The vertebrate kidney has an innate ability for recovery and regeneration following acute injury. andMolecularGenetics,Microbiology SchoolofMedicine, University ofPittsburgh USA C. CiancioloCosentinoandN.Hukriede Kidney DamageandRegeneration inZebrafish Larvae Regeneration and lhx1a , two genes known to play key regulatory roles during kidney embryonic kidney during roles regulatory key play to known genes two pax2a, and Tg(lhx1a:eGFP) ie w sw reactivation. saw we lines Tg(pax2a:eGFP) 354 425

Posters Posters 426 the proximityofroof-platedefinesnichewherethesestemcellscanpersist. and OT the of cells stem neural the contains cells of population this Wethat cells. hypothesize gain-of and loss-of-function of function experiments, combination we show a that WithWnt signalling is signals. essential for Wnt cell-cycle progression of in these source likely the is which roof-plate, mesencephalic the abut and character their in epithelial are cells OT.These dorsal the in population cell proliferative Wnt-responsive, a of existence the for Weevidence provide neural stemcellmaintenanceandpost-embryonicneurogenesis. zone retains proliferating cells during the whole life of the fish, providing a good model to study different cell types. The continuous post-embryonic growth of the OT shows that its germinative of the mesencephalic alar plate into a complex, multilayered structure, containing at least eleven During the embryonic development of teleost fish, it develops from the simple neuroepithelium The optic tectum (OT) is one of the primary processing centers of visual information in vertebrates. Cell andDevelopemntal Biology, University College London, UK M. Varga, R. Young andS.W. Wilson Canonical Wnt SignallingintheDevelopment oftheZebrafish Optic Tectum 355 Regeneration We thank Drs. D. Meyer and H. Okamoto for transgenic fish lines and Dr Mike Tyers for Tyers DrMike and lines fish BioQuarter grant. transgenic for Okamoto H. and providing compounds. This Meyer work is supported by D. a College of Drs. Medicine and Veterinary thank Medicine We the developmentoftherapeuticapproachestomotorneurondiseases. neuron differentiation in development and adult regeneration. This may ultimately contribute to have started to characterise the signalling pathways on which these compounds act during motor We have identified a number ofneurons. smallmotor moleculesborn thatlate influenceof motordifferentiation neuronaccelerate differentiation,or and retard compounds whether determine to fish islet-1:GFPtransgenic Wein embryos. compounds HB9:GFPtransgenic these testing in are outgrowth axonal alter ‘hits’ that molecule small identified have We screens. drug in (GFP) use we compounds, such find transgenic to embryonic order zebrafishIn containing motorcells. neuronesstem thatfrom express greendifferentiation fluorescentneuron protein motor drive in stem cell therapy of motor neuron diseases and there is a need for novel small molecules that motor neurone diseases, and it remains a mystery as to why not. Small molecules could be useful (Reimer et al., 2008, J Neurosci 28:8510-8516). However, humans cannot do regeneration neuron this, motor of capable for are zebrafish example adult that Recently,demonstrated in have we Research Council HumanGeneticsUnit, University ofEdinburgh, UK G. Becker A smallmoleculescreenformotorneuronregeneration inzebrafish Regeneration 1 Centre for Neuroregeneration, University of Edinburgh, Summerhall, Edinburgh, UK;Summerhall,Edinburgh,Edinburgh, of UniversityNeuroregeneration, for Centre 1 , T. Becker 1 , A. Norris 1 , M.Reimer 1 , P. Rengtved-Lundegaard 356 2 , E.Patton 2 2 Medical 427

Posters Posters replacement therapies in humans. This research is being supported by NIH grant DE018043 grant NIH by (PCY). supported being is research This humans. in therapies replacement molecular-based,tissue relevant, mineralized clinically educated, of development the facilitate mediating mineralized skeletal tissues, and replacement tooth formation, which may eventually networks signaling molecular of elucidation the in result will zebrafish in studies proposed the in tissues, mineralized in defects, developmental both late homozygous recessive relatively and heterozygous mutants, on exhibiting viable defects. focuses We it expect that human that to in relevant pathologies disease, identify to geared performed and novel, screen is pilot screen Our laboratory.an earlier this by in identified mutants novel of characterizations continuing 27 F3 families, six of which exhibit early lethal homozygous recessive phenotypes. We are also ENU genotypes, are weekly currently being screened for mineralized tissue performed defects. To Wedate, sibling wt and heterozygous, recessive, homozygous including families, F3 we date. to families F2 have screened teeth. replacement 1,084 individual and F1s, each of which was crossed to wild type AB fish to generate 131 individual primary both mutagenesis of ten males exhibit per week followed by out-crossing and each mutagenized male, to generate skeleton, wild that fact the red on based Alizarin type specimens of this size length, have formed all of the skeletal elements of the axial optimized and craniofacial in ~15mm-25mm an zebrafish of juvenile for consists protocol screen staining Our phenotypes. tissue mineralized exhibiting mutants zebrafish novel identify to screen mutagenesis chemical genetic, forward a performing currently Weare process. this in participating agenes identify to providing model and lives, opportunity unique their throughout formation tooth replacement continuous exhibit Zebrafish 428 C. Stewart-Swift Skeletal MutantsinZebrafish Replacement for ForwardScreen Mutagenesis Chemical ENU Tooth,Genetic Craniofacialand 1 Albertson Tufts University, Boston, USA; Divison of Craniofacial and Molecular Genetics, Department of Oral and Maxillofacial Pathology, 357 2 , P. C. Yelick 1 , M. Connolly 1 1 2 , Y. Lee Biology Research Lab, SyracuseUniversity, Syracuse, USA 1 , D. Fraher 1 , A. Baumbach 1 , J. Chau 1 , D. Sullivan, R. Craig Regeneration 1 1 Analysis ofgeneexpressionfollowing acutehypoxic stressinzebrafish adulthearts Regeneration shmc aae ad ih sget mcaim f ifrnito o cric progenitors cardiac of differentiation of mechanism a present intheadultheart. suggest localized of might repair and the in damages involved be ischemic might that genes development key other of expression after hours of up-regulation the Weobserved ). (kita marker stemness mammalian a to homologous Adult zebrafish heart expresses genes involved in cardiac development and, at low levels, a gene mechanisms ofinjuryrepair, wefocusedourattentiononthesearch forcardiacprogenitors. set up a treatment to cause acute hypoxic stress in adult zebrafish. In order to elucidate possible To simulate the ischemic injury of myocardial infarction caused by coronary artery occlusion, therapeutic implications. we heart. Therefore, to elucidate mechanisms of cardiac regeneration/repair would have interesting model system thanks myocardial to its ability to of replace lost or damaged case tissue, including fin, In spinal cordmuscle and undergoes present. pathologic hypertrophy. are Zebrafish has cells recently become lineage-commited aremaining the while tissue, scar non-contractile by popularreplaced is myocardium infarction,necrotic early regeneration and cells stem containing niches cardiac injury,although severe after repair to capacity little has heart adult Mammalian Dermopatico dell’Immacolata, Roma, Italy Università degli Studi di Milano, Italy; M. Marai, 2 zebrafish exposure to hypoxia and we are currently analyzing possible differential possible analyzing currently are we and hypoxia to exposure zebrafish V. Parente, 2 O. Pozzoli, 2 G. Pompilio, 2 Centro Cardiologico Monzino, Milano, Italy; 3 M. C.Capogrossi, 358 1 F. Cotelli 3 IDI, Istituto 18 kita 429

Posters Posters hnuuk, oy, Japan. Tokyo, Shinjuku-ku, Institute, The Institute of Physical and Chemical Research (RIKEN),Research Chemical Physical and of Institute, Institute The 430 H. Tanaka Expression profilingofneural precursorcellmarkersintheadultzebrafish optictectum 1 deep layer of PGZ. These regions are distinguished by the differential expression patterns of as patterns such genes, expression marker differential cell neuronal the or by stem/precursor neural distinguished are regions These PGZ. of layer deep and superficial as designate we which regions non-mitotic two and region mitotic one regions, distinct three into divided largely is tectum optic of PGZ the that found zebrafish. We adult of PGZ tectum optic the in labeling (BrdU) bromodeoxiuridine with along markers cell precursor stem/ neural known of patterns expression examined we is point, cell this Toprecursor confirm gray unknown. or still stem periventricular neural of of are properties margin have caudal cells cells these the zebrafish whether However, these in adult (PGZ). zone distributed of while are tectum cells area optic proliferating the broad of In most brain. in brain, mammalian observed in regions are restricted the cells in proliferating located brain, teleost adult In Japan not differentiateandmaintainneuralstem/progenitor-like propertiesintheadultoptictectum. progenitor cells the differentiated of into most cells that consisting mature suggest PGZ. findings However,layer.These deep some into populations incorporated may cells these cells, positive elavl3-negative and maintained expression of PGZ. Interestingly, some populations of BrdU-positive cells which ceased cell proliferation were the of most after BrdU incorporation. Then, these cells differentiated into the cells in the superficial layer of strong that showed found and also We cycle incorporation. cell BrdU exited after cells months 2 in region mitotic the in observed still were cells BrdU-positive few that found We incorporation. BrdU after months 2 and elavl3 msi1, eatet f ie cec ad eia BoSine Wsd University, Waseda Bio-Science, Medical and Science Life of Department 359 1,2 , Y. Ito . We also analyzed cell linage of proliferating cells from 24 hours to hours 24 from cells proliferating of linage cell analyzed also We . neurod 1 , H.Okamoto 2 aoaoy o Dvlpetl ee euain Ban Science Brain Regulation, Gene Developmental for Laboratory 1,2 and T. Ohshima xrsin ewe 2 ek ad month 1 and weeks 2 between expression elavl3 sox2, 1 fabp7a, cntfr and pcna, 2-1 msi1. In contrast to Hirosawa,Wako, Saitama, gfap, sox2, 2-2 Regeneration Wakamatsu-cho, fabp7a, elavl3- cntfr, a b hlfl n naeig h cmlxt ivle i te eoee aneac (r lack (or maintenance telomere the in involved thereof) inotherspeciessuchthemouseorhuman. complexity the unraveling in helpful be may regenerative capacity. The study of of the zebrafish’s retention ability the to maintain that lifelong telomerase activity speculate tremendous organism a an with as it as zebrafish the in We necessary is activity telomerase lifelong observed. was telomeres of shortening no clipping, fin by performed regeneration limb following Even points. time all at activity telomerase of amounts abundant and similar expressed also examined tissues All organs. between lengths telomere in difference little was there organ. addition, any In in zebrafish the of lifespan the though shorten appreciably assay.TRAP not and do analysis telomeres Weblot that Southern find by old years Heart, life. adult its of stages through gills, kidney, spleen, activity liver, and intestine telomerase were evaluated at 3 months, 6 months, 9 and months, and 2 length telomere both to regard of its adult life. We have looked systematically at several individual organs of the zebrafish with In sequenced. and cloned much for activity been telomerase maintain zebrafish the that shown been has has it humans, to contrast gene telomerase mammalian the of homolog teleost the biology. of Recently, aspects , ztert many studying for model Marrow growing rapidly a is zebrafish The and Hematology/Oncology/Blood Pediatric of Limb Division Transplant, Minneapolis, or Minnesota, USA Age of Either University by Unaffected T. are C.Lund,T. J.Glass,Tolar, B.R.Blazar Length Telomere Regeneration indaniorerio and Telomerase of Expression Regeneration 360 431

Posters Posters Belmonte new insights into the biology of heart regeneration in the zebrafish and, together with future with together and, zebrafish the experiments inmammals,maybeofpotentialinterestforclinical applications. in offer regeneration heart Ourresults of biology cells. the into epicardial insights and/or new cardiomyocytes zebrafish of cultures primary using specific The role heart. of these new the transcripts during zebrafish heartof regeneration was furtherarea investigated ex vivo regenerating the in upregulated be to shown and hybridization quantitative RT-PCR. For a and/or subset of conventional these by genes, their expression data expression pattern microarray was regulated the analyzed by validated in situ We differentially regeneration. heart expression zebrafish with during genes differential additional the identified ourresults confirmed and analyses transcripts, several Our and compared of data. published regeneration previously heart of zebrafish those of with points time early the on focusing to value potential of be may that treat human cardiac disease. Here, we performed strategies a genome-wide transcriptome profile analysis devise and regeneration heart understand to tool each of role relative the cardiac cell is in this what process still and need response to be regenerative answered, the the zebrafish isdrive emergingthat as asignals very valuable early the are in this process, some of them being cardiomyocyte mitogens in vitro. Though questions as what participate to shown been have molecules and genes several Nonetheless, cells. progenitor of set uncharacterised an from or cells contractile existing from arise cardiomyocytes new if clear an yet not is it evidence, tracing tissue display lineage genetic after of lack the tissue to Due amputation. cardiac ventricular and contrast, partial lost completely restore in deposition to able are Zebrafish, collagen they as capacity proliferation. featuring regenerative the extensive cardiomyocyte episode, tissue infarction negligible scar myocardial with by a remodelling replaced after is mammals, myocardium In available. damaged currently not is therapy Ischemic cardiopathy is the leading cause of death in the world, for which efficient regenerative Institute forBiologicalStudies, La Jolla, USA 432 Sleep E. Transcriptomics approach toinvestigate zebrafish heartregeneration 1 Center for Regenerative Medicine in Barcelona, Barcelona, Spain;Barcelona, Barcelona, in Medicine Regenerative for Center 361 1,3 1,4 S Boué S. , 1,3 C Fernández C. , 1 M Raya M. , 1 Y Richaud Y. , 1,3 Á Raya Á. , 2 ICREA; 3 CIBER-BBN; Regeneration 1,2,3 JC Izpisúa J.C. , 4 Salk 80% (normoxic control; C) O Results: and Methods injury. after occurs response regenerative myocardial a eventually, whether and, heart zebrafish adult Objective: It was examined whether systemic hypoxia followed by reoxygenation damages the ventricular apex.Themechanismsforthisresponsearepoorlycharacterized. Background: –IRCCS,dell’Immacolata Rome, Italy Italy; ne te odtos f h peet td ssei hpxa olwd by in followed marked decrease hypoxia a systemic with study to cardiacregenerationandrepair. associated present damage the lead cardiac may contractility.DNAwhich ventricular in synthesis increase Further,an observed was it of significant induces conditions reoxygenation the Under Conclusions: nuclei were9.20%ofallheartvs0.82%inC. V. Parente Hypoxia andreoxygenationinjuryresponseinthezebrafish adultheart Regeneration 1 determined by BrdU immunofluorescence as an indicator of cell proliferation; 24h after H BrdU DNAcardiac response regenerative a triggered injury reoxygenation and hypoxia was synthesis systolic plane annular valvular by excursion determined as function in decrease significant a exhibited (High-frequence echocardiography by MHz,Vevo77070 System, Ultrasounds VisualSonics) (n=5) H assessed to exposed Zebrafish H. after 16h was function cardiac Finally, group). each in of the stress response gene heme oxygenase1 (hmox1) and of the erythropoietin (epo) gene (n=3 found a 2-6–fold upregulation of genes involved in was apoptosis It (bax, H. after 3-6h heart whole the from RT-PCRisolated RNAquantitative on by (n=2 determined was H death cell after programmed in 18h involved genes and at different of 14 detected expression The upregulated point). was strongly time each it was at and hearts, H C after in 6h absent level wasexpression low caspase-3 immunoblot very activated by evaluated heart; was whole activation the Caspase-3 of p<0.05). analysis group; each in (n=5 C in than higher 6-fold were fragments DNA histone-associated H after 14h apoptosis; DNAduring occurring genomic of degradation internucleosomal assay to performed was lysates tissue cardiac of fraction cytoplasmic the in (n=2 oligo-nucleosomes and mono- C of Determination vs group). each nuclei in apoptotic of number the serial in increase heart 5-fold a of was there staining H after TUNEL 12h sections; by measured was fragmentation DNA Apoptotic treatment. H after 24h and 3h between points time different at analysis for removed was heart the and water Centro Cardiologico Monzino - IRCCS, Milan, Italy; Milan,IRCCS, - Monzino Cardiologico Centro 3 Divisione di Cardiologia,di S.Divisione Ospedale Borromeo, Carlo Milan, Italy; 1 vs C (from 0.71±0.1mm to 0.55±0.1mm , n=5, p<0.05). In order to investigate whether , M.Marai The adult zebrafish heart is known to regenerate following amputation ofthe amputation following toregenerate is known heart zebrafish adult The 2 , L.Squadroni Wild type (WT) adult zebrafish were kept either in 5% (hypoxic; H) or in or H) (hypoxic; Wild 5% in either kept were zebrafish adult (WT) type 2 fish water for 15’. Thereafter H fishes were returned to normoxic 3 , F. Cotelli 2 , G.Pompilio 2 Università degli Studi di Milano, Milan, Milano, di Studi degli Università 1 , O.Pozzoli bcl-2, 362 bcl2l10, 1 4 , M.C.Capogrossi Istituto Dermopatico Istituto bik and api-5), 433 4 +

Posters Posters 434 that Prok2maybeinvolvedinbothadultneurogenesisandpost-lesionneuronalregeneration. suggest Takenresults together,regeneration. our neuronal parallels transcription prok2 ectopic was no longer detected when the wound healing process of was close to transcription completion, showing that Ectopic sites. injury to ventricles telencephalon of regions cortical from of appearance thetransient extending rows weobserved cell broad in later days few a and lesion, the around regions in cells expressing injury, following days first sites. very injury the towards neurons During generated newly of migration the by followed was proliferation this that suggest strongly also data Our sites. injury to distal that and adjacent both located observed regions we zebrafish, adult in in hemisphere injured injury the within proliferation cell in increase dramatic a induced telencephalon lesions brain of paradigm novel a Using brain. zebrafish adult in areas proliferating all almost in neurons to restricted mainly was transcription is not yet fully understood. First, we characterized the zebrafish olfactory bulb neurogenesis in adult mammals. However,including processes biological diverse regulates that protein secreted a is (Prok2) 2 Prokineticin its precise function in of distribution adult the neurogenesis analyse to prok2 transcripts feature in adult this zebrafish brain andused duringWe injury-induced telencephalon regeneration. life. their throughout regions brain several in activity proliferative intense an display zebrafish, including fish, teleost that known well is It CNRS I.C.M.), R (C zebrafish épinière Moelle adult la de et injured Cerveau UMR7225, du InsermUMR-S975, CHUPitié-Salpêtrière, of Paris l’Institut cedex, France de Recherche repair de Centre neural with B. Ayari associated , N.Soussi-Yanicostas andC. Yanicostas is telencephalon expression 2 Prokineticin 363 gene prok2 and showed that its Regeneration - prok2 prok2 are establishingtransgeniclinestostudytheNotchsignaling loss-of-functioneffects. we Additionally staining. antibody and in-situ as well as histology by phenotype this of details various developmental processes. We are currently analyzing the morphological and molecular in cells progenitor other on exert to known also is pathway this that function a cells, blastemal of differentiation delays and proliferation enhances signaling Notch/Delta that hypothesise we Thus, situation. gain-of-function Notch the in tissue differentiated of cost the at cells blastemal control. These preliminary results controlling suggest a prolonged maintenance the or even in ectopic of induction of pattern function expression a the marker blastemal Supportingly has cells. blastema signaling of Notch differentiation and that proliferation indicating outgrowth, regenerative with and formation of the progenitor cells of the blastema has taken place is still sufficient to interfere that Notch found gain-of-function we inhibits (N1aCD) findomain regeneration.intracellular N1aCDNotch1a overexpressionthe afteroverexpress woundto healing line heatshock-inducible a Using regeneration. fin zebrafish in signaling Notch about known to is However, nothing cells. of knowledge hair our regeneration line lateral the zebrafish for and cells required satellite be mouse epithelium, to tracheal and rat andhasa tadpoles Xenopus in metazoa regeneration tail across promote to highly conserved is pivotal pathway importance during embryonic development. Previously, Notch signaling has been Notch/Delta found The regeneration. fin pathway signaling several Notch/Delta pathway components as being regulated by Wnt/ß-catenin signaling during Wnt/ß-catenin of profiling canonical basedtranscriptional the microarray regenerating fins Using in which that regeneration. we specifically blocked fin or enhanced shown zebrafish Wnt signaling we for have identified recently required is have others and We the regeneratingtissueremainunsolved. been have regeneration in identified and recentlyinvolved still major cell pathways questions migration concerningand genetic stem and and cellbiologicalfactors interactions within several involve differentiation Although cell to patterning. tissue cells, thought somatic is of and dedifferentiation morphogenesis and/or postembryonic of proliferation of capable example are striking fish a is teleost animals adult and in regeneration of ability This amphibians parts. body lost or damaged urodele replacing completely like vertebrates nonmammalian Some University of Technology Dresden, BiotechnologyCenter, Germany B. Grotek,B.Kagermeier-Schenk, G.Weidinger The roleofNotch/Delta signalinginadultfinregeneration Regeneration is massively broadened at six days post amputation as compared to the to compared as amputation post days six at broadened massively is msxb 364 435

Posters Posters is agenethatmarksmusclesatellitecellwhosefunctionimportantforeffectiveregeneration. as well show as that Asb11-/- mutants is less effective in homozygous regeneration in response Asb11-/-to muscle injury. wild-types, Therefore, Asb11mutants with gain-of-function of canonical Wnt signalling in at various time-points post-injury, we responses comparing By response. injury in the dorsal flank of the zebrafish muscle, and observed the proliferative and regenerative We show that Asb11 co-localizes with BrdU long term label retaining cells. We induced muscle Here, M-cadherin. quiescent we identifiedmolecule a novel muscle satelliteadhesion cell marker, in the ankyrin and SOCSand box containing identified CD34 11 (Asb11).saliomucin been Pax7, inmuscle as such have cells, satellite markers role molecular Various their precursors. fulfill To myoblast disease.produce to state quiescent or their from activated injury are cells satellite repair, following and hypertrophy regenerationmaintenance, adult of as growth well and formation as in involved muscle, are that cells progenitor are cells satellite Muscle Groningen, University ofGroningen, The Netherlands 436 J-M. Tee Asb11 isamusclesatellitecellmarkerimportantforregeneration etr teh, h Netherlands; The Utrecht, Center 1 Hubrecht Institute for Developmental Biology and Stem Cell Research and University Medical University and Research Cell Stem and Biology Developmental for Institute Hubrecht 365 1 , A. Brouwers 1 , P. vanTijn 1 , M.Peppelenbosch 2 eatet f el ilg, nvriy eia Center Medical University Biology, Cell of Department 2 andD.Zivkovic 1 Regeneration our understanding of this gene family which are emerging as key factors in early embryonic early in factors key as emerging development. are which family toexpand gene this iscritical of understanding our interactions microRNA-mRNA specific of Determination signalling. and to mechanism ideal specification correct allowing cells an to exposure Hh of gradient represents expression precise the maintain pathway developmental critical this of control MicroRNA a cyclopia, muscle fibres. mild and curvature has ventral been indicated by an increase in ptc1 size, expression and increased numbers of superficial slow- embryo reduced phenotype which mimics misregulation of the somites, Hedgehog pathway. Upregulation of the pathway U-shaped in result genes and expression in altered microRNAs when that microRNAs between pathway.Weidentified Hedgehog the interactions in involved and function expression, the evaluate to system model thespecific a as zebrafish the knownconcerning using problem is this approached We microRNAs. individual little of targets investigation extensive despite However field. this of Since their discovery in 1993 the zebrafish identification model has played a pivotal role target in the understanding makes This alignment. the much morecomplex. in can bulges and and 100% not gaps is mRNA mismatches, target demonstrate its and microRNA a between binding of level the species at the post-transcriptional level to downregulate the expression of target mRNAs. In mammalian organogenesis. MicroRNAs are small RNA genes between 19-25 nucleotides in length, which and signalling cell specification, act fate cell including processes cellular multiple in demonstrated The importance of microRNAs in development is now widely accepted and key roles have been Institute ofGenetics, University ofNottingham, UnitedKingdom A. Ketley, E.Holmes,JD.Brook MicroRNA regulationoftheHedgehogpathway Micro RNA andNon–CodingRNAs 366 437

Posters Posters Biophysics Adriano Buzzati Traverso CNR, Italy; 1 438 I. Conte miR-204 isrequiredforvertebrate eye development viaMeis2targetingandPax6 regulation y dvlpet n oe nw vne o a etr opeeso o te pathogenetic the of comprehension better a on avenues new mechanisms underlyingeyedevelopmentaldisorders. open and development eye the in role pivotal the first a time that a specific plays microRNA is and involved in the regulation miR-204 of basic processes of underlying target key vivo generation of this phenotype via a the in regulation of the Pax6 pathway. is and These data demonstrate for Meis2 vitro that in of found variety we a approaches, Through coloboma. and differentiation cell retinal incorrect inmedaka injections bymorpholino formation, lens aberrant microphthalmia, by characterized phenotype function eye severe a determined of miR-204 ablation Targeted fish. medaka development in eye of aspects multiple regulate to capable is miR-204, microRNA, single a that events involved in normal eye development in vertebrates is still largely unknown. Here we The show functional role of specific microRNAs in controlling the morphogenetic and cell differentiation Instituto Cajal, CSIC, Madrid, Spain Telethon Institute of Genetics and Medicine (TIGEM), Naples, Italy;Naples,(TIGEM), Medicine and Genetics of TelethonInstitute 367 1,2 , S.Carrella 1 , R. Avellino 1 , M.Karali 1 Micro RNA andNon–CodingRNAs , R.Marco Ferreres 3 Departamento Departamento de Neurobiología del Desarrollo, 3 , P. Bovolenta 2 Institute of Genetics and Genetics of Institute 3 andS.Banfi 1 works on dazl mRNA itself, the other PGC-specific mRNA. These findings indicate that Dazl that indicate findings ensures PGCspecificgeneexpressionthroughblockingmiRNA-mediatedsilencing. These mRNA. PGC-specific other the itself, mRNA dazl on works through binding to TDRD7 3’UTR and stabilizes the mRNA. We also reveal that this mechanism protein relieve the miR-430 mediated repression of hand, other abundant most the of one cells, microRNA somatic (miRNA), miR-430, represses In zebrafish. in PGCs to restricted is and maternally Our previous study showed that development, Expression ofthesemRNAsisconfinedtoPGCsthroughvariousmechanisms. in Early generations. next program. In produce many gene-expression unique species, acquire that and maternally cells somatic supplied from aside cells set mRNAs (PGCs) cells play germ specialized primordial significant roles the in PGC are development. cells Germ 1 Y. Takeda gene silencing miRNA-mediated the blocking through expression gene specific PGC ensures Dazl Zebrafish Micro RNA andNon–CodingRNAs Department ofBiology,Department Kobe University, Japan; 1,2 , Y. Mishima escapes from the repression by miR-430 in PGCs, Here we show that Dazl that show we Here PGCs, in miR-430 by repression the from escapes TDRD7 1 , H.Sakamoto TDRD7 (Tudor-domain-containing protein 7) mRNA is expressed 1 , K.Inoue TDRD7 expression by inducing deadenylation. On the 2 1 JSPS Research Fellow TDRD7 mRNA. Dazl induces polyadenylation 368 439

Posters Posters 440 are tryingtovalidatewhichmir-125 targets, otherthanp53,couldmediatetheseeffects. We are currently trying cells. to the extend of migration these impaired results to to leads Interestingly,other also angiogenic cells cues. models HUVEC in and, guidance mir-125bexpression more of importantly, proper blocking of a lack reflect supernumerary possibly present could rather that vessels extensions intersomitic filopodial The embryos. these of trunk and tail the in between the intersomitic boundaries. Tunel assays suggested that there is no increased apoptosis most embryos, intersomitic vessels fail to sprout from the dorsal aorta and/or to migrate properly mir-125b-injected duplexembryos),circulation stopsandtheyquicklydevelopheartedema.In and look overall good at that particular dose of morpholino. Instead, in mir-125b morphants (or regulator of p53 (Le at moderate doses of morpholino. Although mir-125b has recently been shown to be a negative phenotype interesting an to leads inhibition its as candidate, angiogenic good a be to appeared Mir-125b stage. one-cell at duplexes RNA of injection through miRNA the of form mature the maturation by injection of a morpholino oligonucleotide into fertilized eggs 2) overexpression of 214, mir-132, mir-365, is mir-125b) has sequence been tested angiogenic mature predicted some targets conserved whose have across distant species. The that biological function of these miRNAs miRNAS candidates (mir-204, selected mir- mainly miRNA also We zebrafish human. in conserved for look to analyses thatcould bioinformatic microRNAs some atdiscovering aimed we potentially be involved in model, regulating angiogenesis. animal As a first as step to our zebrafish study, the we haveUsing performed protein-coding genesandarefrequentlymisregulatedinnumeroustumorigenicprocesses. all of 30% than more regulate to thought are They RNAs. messenger of translation and stability MicroRNAs (miRNAs) are short non coding RNAs shown to exert essential roles by regulating the Giga-R, etdegéniegénétique, unitédebiologie moléculaire Liège, Belgium H. Pendeville, O.Nivelles,L.Malvaux,M.L.Voz, J.A.MartialandI.Struman Finding miRNAs abletoregulateangiogenesis 369 et al., 2009), our morphant embryos display no obvious apoptosis in the head Micro RNA andNon–CodingRNAs in vivo by two means: 1) blocking of the miRNA comparison withourChIP-chipdata. a and binding genomic Eomesodermin and methodology,tail ChIP-seq No for data preliminary coverage of the genome and no loss of data between genome releases. Here we sequencing will present our parallel massively (ChIP-seq). by ChIP-seq data have numerous advantages over microarray data including increased followed immunoprecipitation chromatin is us to available now alternative superior A microarrays. available the of coverage the by limited are they data associated DNA on genomic microarrays (ChIP-chip). Whilst such studies have yielded valuable factor-transcription of analysis by followed immunoprecipitation chromatin using problem this factors regulate development it is important the to expression of specificidentify genes. their Totargets. understand theWe biological repress or activate mechanism either which factors, transcription specific byand general of action the requires whichhave transcription previously determination fate Correct approached types. tissue and cell specialized to leading restricted progressively Embryonic development is a process during which the potential fate of individual cells becomes ofPhysiology,Department Development andNeuroscience, University ofCambridge, UK A. C.Nelson,andF. C.Wardle A ChIP-seqapproach toidentifytargetsof T-box factorsinearlyzebrafish development Genomics 370 441

Posters Posters through NBRP Medakawebsite(http://www.shigen.nig.ac.jp/medaka/). whose transcripts are relatively abundant. All data and genes, clones the in are site searchablestart transcription and of variation open the Wefor observed %. public 12.5 only is cDNAlibrary liver library.in each sequences in unique unique of are Number brain and organogenesisstage whole sequences of each representative clone. About 30 % of clones sequencing from cDNA now are libraries of 3’end. We the of difference the with clones unique 18000 over identified we clones from each library and sequenced both end of the clones. After mass alignment of all data, and ovary) with V-capping method (Kato et. al., DNA Res, seven 12:53-62, now 2006). constructed We are have full-length picked WecDNAup we libraries 24960 project. (early embryogenesis, organogenesis, situation, sequencing fry stage, cDNAmale this liver,full-length overcomebrain, Totestis scale large limited. the very conducting is protein and cDNA medaka as such browser and UT genome browser, the annotation of medaka genome using species specific data Although medaka genome sequences are now publicly available through Ensembl, USCS genome University of Tokyo and K. Ohishi 442 Naruse K. Establishment ofmedakafull-lengthcDNA resources -Anactivity ofNBRPMedaka- 1 ainl nttt fr ai Biology, Basic for Institute National 371 3 , A. Toyoda 1 Y Yoshimura Y. , 3 , T. Aizu 3 National InstituteofGenetics,Japan 1 X Shen X. , 3 , T. Watanabe 1 T Okubo T. , 2 rdae col fArclua ad ie cecs The Sciences, Life and Agricultural of School Graduate 3 , Y. Yamazaki 2 T Shin-I T. , 3 3 , A. Fujiyama Y Minakuchi Y. , 3 and Y. Kohara 3 H Kagoshima H. , Genomics 3 3 , F the central nervous system, notochord, circulatory system pronephric ducts, liver, and intestine. in progeny of injected founders was found in multiple cell types during embryogenesis, including find that founders, on average, harbor 3-4 inserts. Expression of the gene trap fluorescent reporter insertion cassettes, and identified one which gave a germ-line insertion frequency of ~24%. We Ds several of efficacy the tested (Emelyanov,We 2006). zebrafish in transmission germ-line of pilot a performed we 2007-2008, year screen the using maize In cassette. deleter an reporter, a gene-trap and a reporter,contains enhancer-trap and multifunctional, is using are we system transposon The the using are We Emelyanov, A. Temasek Balasubramanian, LifeSciencesLaboratory, 1Research K. Link, NUS, Singapore Ramanathan, A. transposon Hua, R. Ac/Ds S. Parinov Quach, the B. N. using H. Sampath, Zebrafish K. in Screen Mutagenesis system Insertional An FISHTRAP: Genomics neto lns band y hra Aymti Itrae TI) C. e ae performed have We PCR. (TAIL) InterLace Asymmetric F of characterization phenotypic Thermal by obtained lines insertion cen il e ae vial t te agr eerh omnt i de ore ad a the has and course, due in community research larger potential makeasignificantimpactinthefieldofzebrafishfunctionalgenomics. the to available FISHTRAP made the be from will insertions screen Ds The Singapore. in laboratories zebrafish multiple involving We have now embarked upon a large-scale functional genomics screen as a collaborative effort Ds canbeusedsuccessfullytotrapgenesinzebrafish. digestive organs, CNS, heart etc… Our pilot gene trap insertion screen shows that the maize Ac/ 1 progeny of the of progeny insertion lines were confirmed by PCR, and flanking sequences from 74 from sequences flanking and PCR, by confirmed were lines insertion Ds rnpsn o ar ot tasoo isrin cen n zebrafish. in screen insertion transposon a out carry to transposon Ac/Ds Ac/Ds transposon system, which was recently reported to yield in a high rate 2 fish, and have identified insertion mutants with defects in the in defects with mutants insertion identified have and fish, 372 443

Posters Posters 444 aim tofinishfullannotationoftherestgenomewithin thenextfewyears. we and on-going are assembly.strategies current annotation our two to These map that cDNAs 18, 19, 20, 22 & 23 is available in Vega. In 13, addition 10, we 9, have 8, annotation 5, for 4, the 2, majority chromosomes for of path ZFIN clone finished current our of annotation clone Full splices havebeenidentifiedandpreviouslyun-describedgeneshighlighted. alternative further addition, In joined. genes fragmented previously and extended been have models Gene annotation. manual for resource excellent an proven has tissues adult and stages provided a valuable new source of data. Extremely deep coverage from a range of New developmental zebrafish transcriptome sequence from the Illumina (Solexa) Genome Analyzer has recently facilitate theidentificationofnovelsplicevariants,andpoly-A features. the latest species-specific and cross-species cDNA,by ESTprovided and/orevidence protein homologiesby (UniProt)supported and are structures All re-arrangements. and clusters structures, by identified accurately or readily current be automated efforts. Such as, not for example, non-coding may genes, pseudogenes, complex gene that information reliable of wealth a produces clones meticulously finished in-house to an accuracy of over 99.99%. This method of annotation Our manual annotation is based on the reference genomic sequence that has been derived from resource forthezebrafishcommunityasawhole. Network Information Zebrafish the (http://zfin.or with (ZFIN) collaboration close in completed is annotation Our for high quality, Genome frequently Vertebrateupdated, manual the annotation of via vertebrate intervalsfinished genomic(Vega)Annotation regular sequence. (http://vega.sanger.ac.udatabase repository Vegacentral The a k). is database at released is and group (HAVANA) Annotation Wellcome Trust the at entirety its Vertebrateand Human the by provided is annotation manual The Institute. Sanger and in Analysis analysed and sequenced being is genome zebrafish The Wellcome Trust SangerInstitute, Cambridge, UnitedKingdom S. Trevanion, J.Torrance, J.G.Gilbert,L.Harrow, T. Hubbard P.J. Donaldson, S. Laird, K. G. Reimholz, B. Howe, K. Sehra, K. H. Lloyd, M. D. Almeida-King, Havana and Vega: Providing Manual Annotation fortheZebrafish Community 373 ) wih a eald s o rvd a acrt, yai ad distinct and dynamic accurate, an provide to us enabled has which g/), Genomics As the zebrafish is an excellent model for developmental studies, we anticipate thatthe weanticipate studies, fordevelopmental the understandingoffunctiontheseimmunologicalgenes. model improve an excellent will lineages other with comparisons genomic with combined MHC its of elucidation is zebrafish the As following theinitialwholegenomeduplication,vialarge-scale genomicrearrangements. independently several of over each spread other.are assort genes loci This II class and II I class suggests shown has class stickleback in analysis that segregation and the chromosomes, the whereas 11different chromosome teleost on lineages found evolved is independently genes I of region class core orthologous an medaka in teleosts: other in described togetheron been have grouped organisations genes setoforthologous genes. a core 19 chromosome just 19, many other members are found scattered across the contains identified genome. Similar disparate we have contrast, in whilst MHC, zebrafish, In chicken The sub-regions. proteins related three immune 200 within over contained encoding cluster, linked tightly a as exists MHC the cases these in that shown has MHCs dog and pig human, the of annotation and sequencing prior The (DH) fishallowingthereplacementofthesemosaicregionswithasinglehaplotype. genome reference zebrafish the in display major variation. Weto have therefore selected a known tile regions path of BACs from region the double haploid a describing core for conducive not MHC is this haplotypes; The several of consist species. currently of variety wider a from data further for need a is there hence genomics, comparative in point focal a region the makes it remains vertebrates, poorly amongst characterised outside present the of be mammals. to with Its known role is in involved it disease Whilst susceptibility genes and systems. resistance immune of innate and group adaptive a comprises (MHC) complex histocompatibility major The Wellcome Trust SangerInstitute, Hinxton, Cambridge, UK Reimholz, B. S. Trevanion, Howe, D.Stemple,JE.Collins,JLA.Harrow, T. K. Hubbard. Kerry, Donaldson, Auger, G. Storey, KA. RD. S. Griffiths, E. Larbaoui, Lloyd, M. Torrance,Gilbert, W.Chow,J. JGR. DM. Laird, GK. Almeida-King, JP. Sehra, HK. zebrafish Elucidation and comparative analysis o fan MHC haplotype in the CHORI-1073double-haplois Genomics 374 445

Posters Posters 1 1 446 1 the from somatic cellnucleitransfer tonon-enucleateddiploidizedeggs generated transplants nuclear medaka of characteristics clonal of analysis Genetic Resource, Natl. Inst. of Genetics, Mishima, Japan method aretrueclonesofthedonor. new our by produced clones medaka putative the that conclude donor.Wetherefore the from originated transplants three the of nuclei the transplant that indicating HNI-I, the to identical be showed to genotypes obtained polymorphisms the - chromosomes 24 of each on sites four - of one of the transplants. The results showed that, at all of the chromosomal locations examined fins of all three transplants and from the internal organs (brain, eye, liver, kidney, spleen, muscle) DNAanal genomic the and from markers isolated PCR polymorphic 96 using transplants the of the in DNA the of HNI-I strain origin as the donor and the the d-rR strain as identify the recipient. Wemedaka the effectively using therefore transplants assayed nuclear the diploid adult genotypes three to prepared Wesuccessfully transplants. utilized be could polymorphisms these numerous DNA polymorphisms known to exist between them have all been well characterized, belongs strain population. d-rR southern to and population northern of representation the of one is HNI-I population. southern a and population northern a groups, two of consist medaka Japanese of Populations DNA intheirgenome. clone-like transplants, we sought to determine whether the transplants had traces of the recipient and phenotypic traits of donor clones to date. To further clarify the nuclear origin of these donor genotypic the only exhibited have transplants Interestingly,resultant transplants. the nuclear of accepted simply we recipients, as eggs non-enucleated used we Since eggs. diploidized to nuclei of cell somatic adult transfer the through fish medaka fertile and diploid generating for method novel a reported Nuclear transplantation is a key Kyoto, Japan Bioscience and Biotechnology Center, Nagoya University, Nagoya, Japan; Y. Wakamatsu . Adachi, T. 375 1 . Bubenshchikova, E. that the recipient nuclei would contribute, at least in part, to the formation the to part, in least at contribute, would nuclei recipient the that Since these two strains are genetically distinct from each other and the and other each from distinct genetically are strains two these Since technique for cloning animals using somatic cells. We previously 2 . Shinya, M. 3 Graduate school of agriculture, Kyoto University, 3 . Kinoshita, M. 1 . Sawatari, E. 2 Model Fish Genomics 1 . Hashimoto, H. Genomics I. Genome-wide functionalscreenfortheenhancersinzebrafish Genomics of thetissue-specificenhancersinvertebrates. represent time first the for data Our plate. floor and plate roof the particularly in structures, midline the in expression GFP caused insertions where lines ET those for regions of annotation and mapping fine the done have We groups. synexpression tissue-specific different Importantly, a in with some lines lines the reporter gene, ET localized on 220 different chromosomes, of formed distinct collection expression of the reporter gene new and analyzed a genome-wide distribution of a generate to able we strategy, this been Using have line. donor a into mRNA transposase of injection after copyof location new a thegenomic that demonstrated and fish medaka from We have previously established several enhancer trap (ET) lines using gene monitoring for activity despiteonthedifficultiesofdirectidentificationregulatedgene. approach effective the is technique in non-coding trap andenhancer The in by downstream assays. enhancers functional of upstream, distances identification located the largedifficult be makes distribution can over scattered Such they regions. scattered intronic regions coding be of can respect In enhancers sequences. the genome vertebrate In Developmental Biology, IMCB, Singapore, Singapore Kondrychyn , M.Garcia-Lecea, V. Korzh view of genome-wide activity genome-wide of view vivo in a b rmblzd into remobilized be can Tol2 376 Tol2 transposable element Tol2 reintegrations. 447

Posters Posters 448 and (ganglion) thinner. Immunohistochemistry indicates that markers of the inner retina (5E11 (amacrine), Zn5 appears layer photoreceptor the and vacuoles numerous contains and paler is (RPE) epithelium pigment retinal The niche. cell progenitor retinal the is which (CMZ) zone marginal ciliary the dpf 5 to restricted smaller.apopotosis are predominant At eyes the the is Aalthough phenotype striking siblings. to compared shorter Results: by characterised were immunohistochemistry usingzpr1, zpr3,markers 5E11, Zn5andCralbpantibodies. type cell retinal and characterised lamination was Cellular microscopy.morphology light Retinal by function. visual disrupting mutation recessive a with Methods: Optokinetic response (OKR) screens of ENU-mutagenised zebrafish identified a family Aim: To characterisethephenotypesthataffectvisualfunctionindye University Institute,Conway UCD College Dublin, Ireland and Science Biomedical and Biomolecular of School UCD L. Shine,B.Sapetto-Rebow, Y. Alvarez, S. McLoughlin,B.N.Kennedy. on edge(dye) Mutant Progenitor Retinal and Apoptosis CharacteriseMorphology Aberrant Photoreceptor The morphology andprogenitorapoptosis. function defect. Preliminary results suggest that this defect is caused by abnormal photoreceptor Conclusions: We have identified and partially characterised a novelzpr1 stainingofphotoreceptorsissignificantlyreducedindye mutants. zebrafish mutant with a visual 377 mutants display a significantly reduced or absent OKR, and are paler and slightly and paler are and OKR, absent or reduced significantly a display mutants dye (Muller)) stain equivalently in mutants and siblings. However siblings. and mutants in equivalently stain (Muller)) cralbp eia hv udroe xesv lamination, extensive undergone have retinas dye mutant. Genomics and zpr3 dying chemical exposure. to adaptation in involved hspb11is that indicates morpholinos hspb11antisense with injected of muscle precursor cells. Furthermore, the increased toxicity of APM in embryos that have been temporal expression in exposed embryos suggests a potential role in protecting the development and (hsbp11, genes three Interestingly PCR. real-time quantitative by genes of subset inhibits a in that organophosphate changes expression an the validated is and analysis microarray APM performed We acetylcholinesterase. (APM). azinphos-methyl to exposure by mode provoked changes the underlying expression gene sublethal elucidate to and embryo zebrafish toxicity the used we fish Here action. chronic of predict to oflong-term genes responsive theprediction identifying for allow used be can Toxicogenomicwould changes. approaches expression gene analysing by chemicals test of effects endpoints, embryo zebrafish morphological the on that exposures. hypothesise relies chronic Weduring observed currently effects to compared assay if sensitivity the low provide However,may which for animals. alternative as adult considered is of It assessment. testing risk health human as well ) rerio as environmental (Danio of zebrafish The pharmaceuticals. and embryo test (DarT) biocides has been of proposed pesticides, toxicity as the model asses chemicals, for to studying methods chemical industrial acceptable impacts – ethically for and the reliable prediction for demand high a is There Leipzig, Germany Helmholtz Centre for Environmental Research - UFZ, Department of Bioanalytical Ecotoxicology, S. Scholz, N.Klüver, K.Scheffler, andP. Renner azinphos-methyl andacetylcholine esterase inhibitors of effect the models: toxicology predictive as embryos zebrafish in Toxicogenomicresponses Genomics ) were similarly regulated by different acetylcholinesterase inhibitors. Their spatio- Their inhibitors. acetylcholinesterase different by regulated similarly were socs3a) 378 pdlim3b 449

Posters Posters 450 for theimmediatefuture. the project has come to its conclusion in June 2009, establishing such a center is now a priority project. This was precluded by the decentralized nature and limited duration of ZF-MODELS. As European resource center which would preserve the stocks and other materials generated in the While the project has been successful in achieving its aims, a major limitation was the lack of a models, beitthroughlabvisits,training,orbyprovidingmaterialsanddata. by European labs accessible to researchers studying specific developmental processes or disease generated resources high-throughput make to was principle guiding the areas, these of each In Ensembl the with results genome assembly. mutagenesis and microarray integrate knock- to effort of an production and routine zebrafish, the out for technology service TILLING microarray a in of establishment advances the blocks), proteomics, regulatoryand genomic of discovery the to screen enhancer-trap(leading large-scale a brain), the on focusing other the a mutations, with adult (one on screens focus mutagenesis collaborative major two included ZF-MODELS spirit, this In fragmentation inEuropeanresearch byencouraginglarge-scale collaborations. 2004 within the European Commission’s Sixth Framework Programme, which aimed The ZF-MODELS to project overcome(Zebrafish Models for Human Development and Disease) was started in Institute of Toxicology andGenetics, Forschungszentrum Karlsruhe, Karlsruhe, Germany R. GeislerandtheZF-MODELSConsortium Zf-Models -alarge-scaleefforttointegrate Europeanzebrafish research 379 Genomics and chicken, but not conserved in zebrafish, maintain their insulator capacity when tested in tested when this model. capacity insulator their maintain zebrafish, in conserved not but chicken, and tissue-specific enhancer activity in this vector. More strikingly, insulators sequences from mouse highly homologous that conserved non-coding sequences from humans and zebrafish demonstrate with enhancer activity retain their we Additionally, stable activity. enhancer in its transgenesis confirm to of tested control positive the remove to transgenic lines if cassette needed, and 5) a Cre-mediated excision cassette to delete the excision sequence being Flipase a 4) lines; transgenic stable the in and embryos injected the as in RFP-marker efficiency transgenesis a of 3) control positive effects; position from promoter minimal 2) the detection; shield activity to sequences enhancer insulator optimal for chosen promoter minimal specific 1) and improvements: sensitive key a several harbors (ZED) vector (Danio detector” zebrafish enhancer the “zebrafish in This activity rerio). enhancer assessing for vector transposon-based Tol2 novel into a translated is genome is the transcribed and can activate genomic sequences in (enhancers) or repress (silencers) contained of gene expression. Here we information describe the cellular function. These cis-regulatory sequences control when, where activity and how much a gene is how the regulatory understanding of for crucial characterization and identification The for Computational Science, ofBergen, University ofBergenUniversity Norway 1 1 andthe transgenesis tool tofacilitate 1 a new functional analysisofcis-regulatoryregionsinzebrafish vector: (ZED) Detection Enhancer Zebrafish Genomics SPAIN; Centro Andaluz de Biologia del Desarrollo (CABD), CSIC-Universidad Pablo de Olavide, Seville, A. Akalin, J. Bessa, 2 Centro Nacional de Biotecnologia (CNB-CSIC), Madrid, SPAIN; 1 J. Tena, 1 B. Lenhard, 1 A. Miñán, 1 F. Casaresand 1 E. de la Calle-Mustienes, 1 J. L.Gómez-Skarmeta* 1 S. Naranjo, 2 A. Fernández, 380 3 Sars And Bergen Center 2,3 L. Montoliu, 451

Posters Posters 3 452 S. Berger Reverse Geneticsin Australia: Fishing fornovel zebrafish mutants Haematology Division, Walter and Eliza Hall Institute of Medical Research,Parkville,Australia;Medical of Institute Hall Eliza and Division,Walter Haematology and theresearcher willreceivethefishforfurtheranalysis. frozen sperm sample will be used for the resurrection of the mutant through researchersIn Vitro Fertilization allow will that samples from the DNAentire community to screen accompanying for mutants of their interest. with After identification of such,samples the sperm frozen 5000 of We will now continue to perform state of the art ENU mutagenesis and fin mutants,twoheartnoveldystrophicandonemusclemutant. will collect a total number F2 families were screened for muscle mutants. This F2 screen resulted in the identification of two The remaining mutant Vitro In F1 via females mutant identified were one crossed Fertilization fromafrozenspermsamplethatwasstoredfor2years. resurrected into we principle, a of Tg(acta1:GFP)zf13 proof as background. Furthermore, The kb. resulting 122 every exchange base-pair 1 of rate mutagenesis a wit mutations of saturation high a confirmed analysis The rate. mutagenesis the calculate to sequenced were genome entire the across genes known of Amplicons collected. was extraction DNA for Simultaneously,tissue freezing. sperm crossed the mutagenised males to wild type females. The resulting mutants were used to perform We have treated wild type males with library thatallowsaccesstomutantsfarbeyondthetimelimitofazebrafishlifespan. either be kept alive during the screening process or can be used to set up a frozen sperm / DNA Requirement for either of those methods is the generation of randomly mutagenised fish that can DNA samples,therebydetectingmutantswithhighthroughputandlowcosts. to applicable be of largegroups of analysis to the allows sequencing Deep 2002). shown (Wienholds al., also et zebrafish was and invertebrates and plants for established Genome) been initially In has Lesions Local (TargetInduced TILLING sequencing. deep reverse and for TILLING available approaches genetics: two currently are There function. its study to out knocked is genes. At the moment, fewer methods are available for reverse genetics, where a gene of interest function in a forward genetic manner, where identified phenotypes lead to the discovery gene investigating on mostly organism relies of model genetic vertebrate a as novel zebrafish of use The Australia 1 Colon Molecular and Cell Biology Department, Ludwig Institute for Cancer Research,Cancer Department,for Parkville,CellBiology LudwigInstitute and Colon Molecular University,Institute,Monash Medicine Regenerative Australian Clayton, Australia; 381 1 , J.Berger 1 , G.JLieschke 2 , J.K.HeathandP.D. Currie N-ethyl--nitrosourea (ENU) and have subsequently out- 1 Genomics 2 Cancer and Cancer The sequencingprojectmovesintomaintenancephaseinJanuary2010. at viewed be includes linkstopre-Ensemblwherethenewassemblyzv8canbefound. can assembly latest The which aidthecoordinationandanalysisofgenomearedescribed. in finishing this genome, and report results from trials using the new technologyfaced platforms.challenges unique the of some of Tools examples present and progress sequencing Wediscuss to helpanchorcontigs,resolveorderandestimategapsizesalongchromosomes. such regions and extend clone coverage. Fluorescence in situ hybridization (FISH) is being used resolve to used been have (CH-73/1073) fish haploid doubled single a from generated libraries Many regions of the genome appear to have high levels of haplotype diversity; BAC and fosmid selection andgapclosure. clone final for basis the forms now map this fish; homozygous 48 from markers of analysis by addition of Whole Genome Shotgun (WGS) data. In 2008, a new Heat Shock map was generated 99.99% accuracy in euchromatic sequence), with the assembly being aided and least informed (at by standard” “gold the to sequenced and mapped fingerprinted, clones, BAC of libraries with began used strategy The world. the single- in 3 largestPhase HTGS Zebrafish to the taken be to the is project sequencing centre this sequence size, to in 1.36Gb project At Trust. year Wellcome the nine by its entirely of funded genome, months final the in is Institute Sanger The Sequencing, Wellcome Trust SangerInstitute, Cambridge, UK S. SimsandL.Matthews The Zebrafish Genome:anUpdateofMappingandSequencingProgress Genomics which www.sanger.ac.uk/Projects/D_rerio/wgs.shtml 382 453

Posters Posters 454 capillarysequencing. next 2years,creatingarichresource forthezebrafishresearch community. by areconfirmed the over alleles knockouts >1000 Resource produce Mutation to Zebrafish aims the method this Using non-sense putative and the in fragments, alleles non-sense amplified potential all for sequence the searches then analysis data Automated platform. Solexa the of lane one on run is pool each and zebrafish, mutagenised 24 from pools by allele non-sense a give ENU to mutagenesis. These likely fragments most (up to are 4000 exons that per experiment) fragments are exonic amplified across100bp DNA selects that written platform. sequencing Solexa AIllumina been the has of fragments program lane design single primer a on exonic of thousands anon- sequence and amplify to us ofidentifying allow will that strategy a thepossibility designed up opened Webudget. have has and frame time reasonable a within gene coding and protein every in allele sense identified be can that mutations amount of the changed significantly has technologies sequencing generation next of advent The ac.uk/Projects/D_rerio/mutres/. Information database). Phenote website: our the on found be of can project the version by identified mutations adapted the about an (using ZFIN via published be Mutation will theZebrafish data by Phenotypic phenotype. molecular and created morphological associated isbeing an with each Resource, alleles non-sense Zebrafish of library A manner. shown that capillary-sequencing can be used to effectively identify gene. any virtually a in identified be to mutations allows that method proven mutations previously have We is in genomes), a in lesions high local throughput induced targeting (for TILLING or mutagenesis, Targetselected Wellcome Trust SangerInstitute, Wellcome Trust GenomeCampus, Hinxton, Cambridge, UK R. Kettleborough; E.Busch-Nentwich;C.Herd;F. Fenyes;C.Torroja; F. vanEeden;D.Stemple The Zebrafish MutationResource: Towards a TILLINGknockout inevery proteincodinggene 383 http://www.sanger. Genomics 2 C. Torroja Integration ofheat-shock, mghandt51geneticmapsintozebrafish genomeassembly(zv8) Genomics 1 closer to the size predicted by flow cytometry. Assignment of sequence to Chromosome 4, toChromosome repetitive sequence,whichcouldleadtomis-localisationof FPCs. very contains 4 ofsequence Chromosome of arm long The Assignment caution. cytometry. with interpreted be flow to however,needs by predicted size the now to size increased closer significantly a has 4 Chromosome that find we each addition, for In 0.96 of chromosome. average an at now is which maps, genetic the of each and assembly Zv8 the maximum the to resolution and coverage possible given available the information. Weincrease see a we striking improvement in maps the correlation between three all using By FPC. each of sequence the and markers genetic between alignment sequence by maps different three the Wemaps. different three from derived data position map integrate to association this used have the between association an makes therefore and FPC the on mapped markers the between link a provides FPC The unit. indivisible an as FPC and an consider we process this order In orientation. and long-range for MGH and HS maps chromosome assignment, then using the T51 radiation hybrid map mostly to resolve local meiotic order the prioritising FPCs, the reorganized genetic maps: Heat Shock (HS), MGH meiotic maps and the T51 radiation hybrid map. We have To improve Zv8 quality we have reorganized FPC order and between FPCsandresolvingthemverydifficult. orientation by combining the existing their map to trying researches for mutations as issue well as for the important genome sequencing project, making the an task of identifying real is gaps This 0.7. around only is assembly the the assembly and the map. Indeed, for Zv7 the average correlation between the three maps and used to anchor physical map fingerprint contigs (FPCs) in Zv7, there is poor correlation between genetic maps (HS, MGH and T51) highlights this problem. Despite the fact that the T51 map markers was represented in these assemblies. Analysis of the correlation between Zv7 and the three there have been many discrepancies between genetic mapping positions and the position of the Due to the choice of genetic map (T51) on which previous Zebrafish assemblies have been built, em1 Wlcm Tut agr nttt, eloe rs Gnm Cmu, ixo, UK; Hinxton, Campus, Genome Team71. Trust Wellcome Trust Wellcome SangerInstitute, Wellcome Institute, Trust GenomeCampus, Hinxton, UK Sanger Trust Wellcome Team31. 1 , J.Torrace 2 , B.Reimholz 2 , K.Howe 2 , D.Stemple 1 384 455

Posters Posters 456 see us! and come Please resources. these of use the regarding problems individual with help also will We advertised). be to (details workshop one-day a in and sessions poster the during resources web available the of use the demonstrate and community the to data new the present Wewill it for annotation displayed inEnsembl. automated and manual integrated provide and assembly only clone finished a create eventually will we completion, its nearing project sequencing sanger.ac.uk).the With (vega. Vegabrowser the in provided is annotation manual with only sequence clone Finished http://www.sanger.ac.uk/cgibin/Projects/D_rerio/ community at is for annotation Annotation/submitAnnotation.pl. and track your pipeline a enter build offers Please gene also Ensembl now annotation. the Ensembl (www.ensembl.org). using Ensembl annotated in fully accessible been has assembly Zv8 The was usedforZv7. than coverage fold greater a with assembly (WGS) shotgun genome whole a from contigs with this In haplotypes map. several and those have of subsequently hybrid been removed. Gaps between the finished clones have use been filled radiation the T51 from arising the duplications and artificial further MGH identified and we process, HS maps genetic oriented the and of ordered use been careful have by clones these and clones finished high-quality of precentage higher a contains It predecessors. its to compared improvement major shows assembly genome The Sanger Institute has recently released a new integrated zebrafish genome assembly, Zv8. This Informatics, Wellcome Trust SangerInstitute, Cambridge, UK K. Howe, W. Chow, B.Reimholz,J.Torrance andT. Hubbard The Zebrafish GenomeSequencingProject: Web resources 385 Genomics 1 O. Konu Meta-analysis ofrapamycin modulatedexpressionprofilesinthezebrafish andhuman Bioinformatics andsystembiology core meta-signature, common to zebrafish and human transcriptomes, could be extracted for extracted defining themolecularcontributorsof TOR signalinhibitionviarapamycin. be could transcriptomes, human and zebrafish to common meta-signature, core were units signaling and performed using genes available tools altered such as commonly DAVIDthe and of Pathway Miner.annotation Functional In datasets. summary, PC3 an orthologous rapamycin modulated gene expression zebrafish signature was compared addition, with those obtained In from public MCF7 embryogenesis. and zebrafish during regulated differentially rapamycin was with genes down-regulated and up- of expression maternal that indicated (www.bioconductor.org).findings Our modules R using representation visual and testing statistical for selected human and zebrafish Affymetrix datasets were made accessible through a web-interface which in tool meta-analysis data. a microarray developed human Affymetrix We and zebrafish gene- of of rapamycin analysis on ZF4 transcriptome; and then meta-analyzed this dataset using comparative a series of public for opportunities expression unique modules. Accordingly, we provides performed a microarray experiment to decipher datasets the effects of multivariate Meta-analysis of humans. such to numbers zebrafish from largeranging arebeing species vertebrate context, conditions multiple for andpathological this generated help In physiological different genomes. studies under profiles the control expression in gene transcriptional components functional and the profiling deciphering gene-expression high-throughput Recent Biotechnology, Ankara University, Turkey BilkentUniversity, Genetics,and Biology Molecular of Department Ankara, Turkey; 1 , C. Sucularli 1 , K.D.Kaya 1 , A. R.Ozturk 1 , H.Ozdag 2 386 2 Institute of Institute 457

Posters Posters system should produce an “estrogen sensitive” adult transgenic fish, but this has yet tobe yet has this in earlylifestageembryostodetectexogenousoestrogenexposure. but fish, transgenic adult sensitive” confirmed. This “estrogen study is an also the produce first to should report onsystem the application of ERE-driving reporter constructs amplification step two our that suggests UAS-GFP and into embryos introduced zebrafish when oestrogen to response a provides produced system plasmid the that confirmed have data These the BPA exposedgroups(concentrationsrangingbetween10µg/L and1mg/L). of any in seen BPAwas ug/L. expression GFP10 enhanced of No exposure an at observed was expression GFP but (100µg/L), tested concentration highest the at died embryos all exposures, NP the For exposure). 24h (after groups treated ngEE2/L) 10 ngE2/L, (100 low and EE2/E2) ng/L (1000 high the both in cells other many and epithelium skin lens, the in detected was GFP of and GFP expression detected in embryos in unexposed controls, (NP) but specific and mosaic expression oestrogen Nonylphenol synthetic estrogens the environmental (E2), Bisphenol weak A (BPA) 17ß-oestradiol and the GFP expression examined using a fluorescent oestrogen microscope. and There was no steroid (EE2), natural 17α-ethynyloestradiol theembryos the and to embryos, exposed zebrafish were protein) fluorescent (green UAS-GFP transgenic staged 1cell into separately embryosinjected were zebrafish (10-20pg) plasmids in assay.The exposure expression transient estrogen a using to construct our of response the examined have We EGFP (EnhancedGreenFluorescentProtein). gene, reporter to second a of expression exposure the drive these on and elements (UAS) Sequence that Activated (3ERE) elements response estrogen estrogen induce of expression of pERE-TATA- copies Gal4FF,plasmid a three this developed in We containing turn system. Gal4FF,binds Gal4-UAS specificallya toof GAL4-responsiveuse Upstream the adopted we to fish, oestrogen, sensitivity response adult the improve in to attempt gene an sensitivity.In reporter limited have Luciferase systems these a but with promoters natural ERE-containing responsive or (estrogen elements) ERE synthesised artificially used has chemicals oestrogenic detecting for work transgenic Previous oestrogen. to responsive highly are that zebrafish transgenic develop to attempting are we water the in (EDCs) chemicals disrupting Toendocrine oestrogenic detect Biology, University College London, UK 458 O. Lee Development ofestrogen-responsive transgenic zebrafish usingaGal4-UASsystem 1 School of Biosciences, University of Exeter,of UK;University Biosciences, of School 387 1 , M.Tada 2 , C.Tyler 1 andT. Kudoh 1 2 Department of Anatomy and Developmental and Anatomy of Department Emerging technologies and otherstudies,weproposerecommendationsfortheuseofMOstoanalyzegenefunction. involving phenotypes we p53, active transcriptional activity,p53 of direct but by are presence instead upregulated induced not the by are a genes in Bcl-dependent expressed mechanism. ectopically it these These that unexpected blocking show non-specific conversely and p53, of downstream pathway the mitochondrial apoptotic pathway is required. By pharmacologically activating the apoptotic that find mechanism surprisingly and expression, the gene in changes examined these Weproduces activation system. p53 which by nervous central developing the in genes of subset a of gene of expression (possibly due loss to cell loss) only but also an not intriguing up-regulation and causes ectopic expression activity p53 of induction non-specific this that show We defects. in expression related gene cell death via p53 of activation, resulting in knockdown loss of embryonic structures for and morphological used tool zebrafish. Robu et al. (2007) demonstrated that a large main fraction of morpholinos cause non-target the are (MOs) morpholinos Antisense National InstituteforMedicalResearch, MillHill, London, UK S. GeretyandD. G. Wilkinson Antisense morpholinotoxicityreveals novel roleforapoptoticgenes Emerging technologies of gene expression add to the pitfalls of using MOs. Based on these on Based MOs. using of pitfalls the to add expression gene of gain 388 459

Posters Posters 460 reagents atwhateverdevelopmentalstageisofinterest. conclusion, by at least 80%, as assessed by ratioing GFP (targeted) to mCherry (not-targeted) fluorescence. In GFPof expression the prevented oligonucleotides anti-GFPRNAi of Inclusion loss-of-function. using of efficacy the determine to used was mCherry GFPand of Co-expression examined. cells the of 95% in mCherry GFPand of co-expression to leads mixture electoroporation the in plasmid expression mCherry an and plasmid expression can also efficiently incorporate two different reagents at the same time – inclusion of both a GFP- method This post-fertilization. hours 96 and 48, 24, including stages developmental multiple at toefficient, lead by 24 hours. expression maximal reaches that and electroporation, of hours voltages 6 within observed that is expression find We retina. reproducible GFP and expression are well within the range for cerebellum, which there is very high viability. hindbrain, GFP midbrain, including neurons developing of populations neuronal multiple target can electroporation that show method. We the of efficacy, resolution the viability, temporal determine and to used was a specific stages in development. characterized a at reagents have loss-of-function incorporate to used we be can which Here electroporation, vivo in development. method, neural of assessment compromise may stage of gene cell the one the at If starting loss-of-function step, system. development to in nervous earlier an for the attempting necessary is interest of when development as problematic such be events developmental can later which study development, throughout present RNAi is and reagent loss-of-function morpholino the that is (e.g. approach this to reagents caveat One stage. loss-of-function loss-of-function cell one the gene of at oligonuceotides) zebrafish, microinjection In by development. achieved of be aspects can various for important are genes which about information of wealth a yielded has organisms model in analysis Loss-of-function Biology, Pace University, Pleasantville, USA J. H.Horne,S. A. Kera,S.M. Agerwala Using In Vivo Electroporation for Time-Resolved GeneticsinZebrafish 389 In vivo electroporation is an effective method for delivery of GFP expression plasmids electroporation can be used to deliver both expression plasmids and RNAi and plasmids expression both deliver to used be can electroporation vivo in In vivo electroporation of GFP and mCherry expression plasmids in vivoin RNAi-mediated for electroporation Emerging technologies to inducegeneexpressioninatinyareaofovary. us allowed successfully method This possible. was cavity,treatment body heat the noninvasive into probe this inserting By monitored. and controlled be can temperature thin whose a probe employed metal Secondly, we fins. and liver a gonads, as such organs some in observed was First, medaka was kept in a hot water (39C or 42C) bath. ToA achieve appropriate conditions of heat treatment in day adult medaka, we attempted two means. after the treatment, GFP expression observed inmedakaembryos,followingbriefheattreatment. loxP construct was injected into the embryos of cre lines, the expression of the reporter gene was the embryos. After medaka in treatment heat We by worked cre-loxP system the that confirmed crossed toeffectorlines(loxP[DsRed]-GFP),whichallowsexpressGFP byheattreatment. with cre gene under the control of heat shock elements (HSEs) were developed. These lines were of various biological phenomena. To achieve the purposes, medaka transgenic lines (driver lines) Induction of gene expression in any organ at any time is an important technique for investigation Japan Laboratory of Molecular Genetics for Reproduction, National Institute for Basic Biology, Okazaki, K. Kobayashi andM. Tanaka Heat InductionoftheGeneinMedakaEmbryos and Adult Medaka Emerging technologies 390 461

Posters Posters 462 I. Rothenaigner Viral celltransduction intheadultzebrafish brain Zebrafish Neurogenetics; 1 oti sl-eeig rgntr el ta so faue o aut amla nua stem neural mammalian adult of followed be features can division cell that show showed experiments tracing BrdU cells. that cells progenitor self-renewing contain ventricle, brain the to close located mostly anterior-posteriorzones, entire germinal These axis. by incorporation of the thymidine analog BrdU, can be found in several brain regions visualized zones, Proliferating cells. The adult zebrafish brain shows in contrast to mammals an enormous potential to produce new dl ban n wl hl t frhr dniy h mlclr ehnss otoln se cell stem controlling mechanisms molecular the identify maintenance anddifferentiation. further to help will zebrafish and live the brain in adult cells progenitor of tracing genetic long-term the permit will method This and glial markers. neuronal of stainings immunohistological using cells infected of and fate term combinations long technical the reveal successful of set and the retroviruses classify of will types experiments two Further with promoters. zones ventricular the along cells Preliminary of zebrafish. infection adult show live results of ventricles brain the into retroviruses GFP-encoding based live adult the the in transgene is stably integrated into the genome of cells the target cells. We injected these MLV- and of HIV- transduction zebrafish virus-mediated brain. established We chose retrovirus-based we systems, which purpose have the this advantage that afterFor infection cells bypermanentlylabellingthem. The aim of our study was now to develop a tool to analyze the long-term postmitotic neurons.fate of neural progenitor Helmholtz Center Munich- German Research Center for Environmental Health, Department Department Health, Environmental for Center Research German Munich- Center Helmholtz 391 1 , R.Jagasia 2 Institute ofPhysiology, University ofMunich 1 , A. Lepier so far with the proliferation markers PCNA and MCM5 or MCM5 and PCNA markers proliferation the with far so 2 , P. Chapouton 1 , B.Bahn 1 , L.Bally-Cuif Emerging technologies by the by 1 formation of formation along the medaka GFP line using xenobiotic inducible xenobiotic using line GFP medaka potentially and metals heavy by developing in interested also are caused we Tg(hsp70:gfp), Besides determined. be to chemicals other stressed monitoring for used be may line transgenic developed newly the Thus cadmium. and mercury,arsenic including metals heavy several to endogenous of rges n w hp t caatrz ti ln wt vros aeois f esset organic persistent of categories various with line this pollutants (POP)includingpolyhalogenatedbiphenyls,pesticides andetc. characterize to in hope is Tg(cyp1a1:gfp) we for and founders progress of screening Currently, pollutants. such monitoring for line and BaP as Tg(cyp1a1:gfp) such stable develop compounds to potential xenobiotic 2,3,7,8-Tetrachlorodibenzo-p-dioxin,the indicating by elevated was expression GFP that the found also under we construct GFP a of injection By control. vehicle to compared (BaP) benzo[a]pyrene to exposed when embryo dpf 6 of liver in elevated at 37 at lines we obtained show strong ofubiquitous GFP induction within 4 hours of heat them, shock Out treatment of system. 10 from transposon transgene Ac/Ds the indicating a of high maize efficiency oftransmission the Ac/Ds of germline system to aidobtained transgene aidintegration.we All of thescreened, the transgenic founders with 12 generated was line transgenic The promoter, stress-inducible a using line medaka Tech.Sci. Eng. al., et 39:9001). (Zeng In compounds the estrogenic current of presentation, surveillance online we for report amendable the tool estrogenic generation other of second and biomonitoring estrogen transgenic to responded compounds faithfully by displaying line green transgenic fluorescence Tg(mvtg1:gfp) color and the thusthat provideddemonstrated and agene convenientvitellogenin biomonitoring from promoter fish estrogen-inducible transgenic the under color line medaka living the of apply transgenic to GFP a generated feasible has laboratory University Previously,our is contamination. water monitoring it in gene, National reporter GFP of Sciences, advent the With Biological of Department and Singapore School Graduate NUG H.B.G. NgandZhiyuanGong Development ofliving colortransgenic medakaforbiomonitoringaquaticcontamination Emerging technologies o C for two hours in 2 dpf or older embryos. The expression of GFP is consistent with that with consistent is GFP of expression The embryos. older or dpf 2 in hours two for C mRNA after heat shock. In addition, the transgenic line also responded also line transgenic the addition, In shock. heat after mRNA hsp70 promoter.cyp1a1 Transcript of level Tg(hsp70:gfp). of designation the with hsp70, rmtr no eaa embryos, medaka into promoter cyp1a1 392 was cyp1a1 463

Posters Posters shared betweendifferentcyclicgenes. cyclic genes such as elements our regulatory test putative and of deletions generating currently are we experiment principle oscillations of protein track to time first the for us allow lines these Additionally patterns. mRNA endogenous the from indistinguishable is activity reporter whose her7::Venus,her1::Venus for and lines transgenic multiple created we approach this on Based enhanced transgenesisfrequencies. modified the of size desired the subcloned Venus, the manipulate to recombineering BAC endogenous employed have we problem this overcome To level. asprotein and mRNA such the both on patterns stability reporter expression prolonged of dynamic because expression very gene cyclic recapitulate to task transgene challenging Conventional the expression. fail gene constructs cyclic of reporters transcriptional reliable generate the and for elements requirement regulatory the putative test these of to sufficiency order In elements. intergenic conserved, highly revealed 464 closely-related both of the footprinting of phylogenetic out clones carried BAC and goldfish orthologous sequenced and isolated Tonarrow have kb). we 12 down, (~ these region intergenic their within elements regulatory share might they implying that orientation, head-to-head a in located and coexpressed spatially and temporally are zebrafish the chose we hypothesis Tothis test and testwhetherdifferentcyclicgenesshareaconservedenhancerarchitecture. expression gene cyclic confer that elements regulatory characterize and identify to and is aim Our clock this of components study are to genes model controlled Cyclic at interesting the level. transcriptional an level. molecular However, as the little emerged is at known about has processes their enhancer clock architecture.oscillatory segmentation the decade last the Over CBG, MPi-CBG, Dresden, Germany D. Soroldoniand A.C. Oates Combining bacrecombineering&I-SCEItransgenesis toassessoscillatorygeneexpression 393 predictions silico in ou wtot ie iiain. e ae agd either tagged have We limitations. size without locus her1/7 dlC, which could eventually shed light on a common enhancer architecture . In parallel we carry out a similar approach for other for approach similar a out carry we parallel In vivo. in locus as starting point. Both cyclic genes cyclic Both point. starting as locus her1/7 , we have introduced a new logic to logic new a introduced have we vivo , in loci and introduced and loci her1/7 loci. This loci. her1/7 Emerging technologies . Based on this proof this on Based vivo . in or her1 approach silico in sites for sites I-SceI with her7 E. Ropstad 3 Nourizadeh-Lillabadi R. Exposure effectsonzebrafish ofnatural POPmixtures Emerging technologies 1 risk tothereproductivehealthofwildfishinhabitatingfreshwatersystem. a represents pollution chemical whether of question the raise model zebrafish experimental the in observed effects The events. those in role main a endocrine having not PBDEs the suggesting mixtures, in involved genes of by Ingenunity Pathway Analysis (IPA) expression indicate similar effects on puberty and the weight gain by both predicted Networks effects. phenotypic in observed the with accordance in growth and signalling changes and liver revealed exposed The age. transcriptomes of months gonad 5 at weight body increased and ratio sex male/female Phenotypic effects observed in both exposure groups include earlier onset of puberty, increased Lake Mjøsa,toconcentrationsreportedinhumanandwildlifepopulations. from fish wild in detected levels from ranged zebrafish the in measured POPs of concentration background levels of PBDE. Both mixtures had low, with but mixture one substantial and levels (PBDEs) ethers of diphenyl PCBs polybrominated of and levels high DDTs.with mixture were one The (POPs) pollutants organic used were extracted from persistent burbot (Lota ) lota liver originating from freshwater systems in Norway: of mixtures natural investigated using classical 2 and molecular methods of in a controlled concentrations zebrafish model. Therelevant mixtures In the present study, developmental and reproductive effects of lifelong exposure to environmental owga Sho o Vtrnr Sine Norwey; Science,Ullevaal University Hospital, Norway Veterinary of School Norwegian 1 1 , J. L. LycheL. J. , 1 , C. Almås C. , 1 , B. Stavik B. , 2 ainl eeiay nttt, Norway; Institute, Veterinary National 3 , V., Berg 394 1 , J. U. Skåre U. J. , 2 , P., Alestrøm 465 1 ,

Posters Posters 466 a combinationofautomatedandmanualmethods. organism, embryos will be screened for morphological, enzymatic and fluorescent output using Towhole a using when available are that methods screening different the of advantage full take multiple pathwaysinthepathogenicprocess. nature of our models provides the potential for identification and validation of reagents affecting organismalwhole the proteins, and targetpathways disease-relevant of number small relatively a on focused are efforts where programs, screening drug conventional to discovery.contrast In drug for compounds lead of validation and identification the speed Moreto systems of establishment screens. molecular content high have biological assayswiththedualaimofidentifyingnovelreagentsforcontinuedresearch andthe for Genetics basis Biomedical a and provide recently, Developmental we that have established for a facility models for medium-throughput Centre screening disease using such MRC high established content the within groups Several The Court, Facility, Firth Screening University ofSheffield, Genetics Western Bank, UK Biomedical and Developmental for Centre MRC S. Baxendale,C.Parkin,SmithandM.Placzek using thezebrafish Screening for small molecule modulators of disease processes and developmental mechanisms 395 Emerging technologies spontaneous slow forward swimming. The responsible neuron is the Kolmer-Agduhr (KA) cell Kolmer-Agduhr(KA) the is neuron responsible The swimming. forward slow spontaneous mimics that sequence beating tail a symmetrical induce to sufficient was type cell specific one of intersectional gene expression and optogenetics CPG that drive spontaneous locomotion. Here we identified such an input using a combination oooin eis n erl ewrs ald eta pten eeaos CG) ht generate that (CPGs) generators pattern periodic motor central commands called for rhythmic networks movements neural on relies Locomotion Berkeley NationalLaboratory, USA that terminates in a series of consecutive segments consecutive of series a in terminates that axon ascending ipsilateral an with cord spinal the of canal central the into cilia extends which 1 C. Wyart that drive locomotion neurons contacting fluid cerebrospinal spinal identifies behaviour of dissection Optogenetic Emerging technologies of California in Berkeley,in California of USA. of California in San Francisco,San USA.in California of vertebrates. in conserved be may system this that and development early in CPG locomotory the for drive positive a provide neurons contacting fluid cerebrospinal spinal that suggest results Our no cord. had connections brain descending of effect on the KA-induced Ablation behaviour, demonstrating that swimming. the KA circuit operates forward within necessary the spinal provides spontaneous activity for cell tone KA that indicating swimming, free spontaneous of frequency Munich,Germany. Helen Wills Neuroscience Institute and Department of Molecular and Cell Biology,Cell University and Molecular of Department and Institute Neuroscience Wills Helen 1 , F. DelBene 4 hscl isine iiin n Mtra Sine iiin Lawrence Division, Science Material and Division Bioscience Physical 2 , E.Warp 2 Department of Physiology,of Department Neuroscience,University in Program 1 , E.K.Scott 3 Department of Chemistry,of Maximilians-Universität, LudwigDepartment 2 , D.Trauner 2 1 4 in zebrafish larvae. The photo-stimulation of . Little is known about the spinal inputs to the . Genetically silencing KA cells reduced the reduced KAcells silencing Genetically . 3 , H.Baier 396 2 , andE. Y. Isacoff 1,4, 467 3 ,

Posters Posters 468 in vertebrateembryos. for investigating the regulation of ECM remodeling in the zebrafish embryo are now unparalleled conjunction with classical embryological, biochemical and molecular techniques, opportunities inthe abundance, and activity patterns of these ECM remodeling effectors homologes in the zebrafish embryo. MMP 23 identified have We zebrafish phenomena. genome and have developed several these techniques and reagents to assay study the distribution, to which in system context of a whole organism. We propose that the zebrafish embryo presents an excellent model due to the complexity of their post-translational regulation, much less work has been done in the understanding the biochemistry and cell biology of MMPs and their regulation, however, largely tumor metastasis, heart disease and arthritis. remodeling ECM for in vertebrates. Misregulation of these proteases is central to a plethora of pathologies, including responsible largely subsequent are ofzinc-dependent - and (MMPs) metalloproteases family matrix matrix complex the A the - configuration. proteases new into a proteases in molecules of ECM secretion of re-synthesis the necessitates healing wound and As a non-living component of multicellular tissues, remodeling of the ECM during development and proteins of array properties. functional and mechanical essential with tissues provide that diverse macromolecules other biochemically a of comprised is (ECM) matrix extracellular The University ofNew Brunswick, Fredericton, Canada B.D. Crawford The zebrafish embryo asamodelforstudying extracellular matrix dynamics 397 As such, a great deal of research has been focused on Emerging technologies In

with either: lines transgenic golden establish to system Tol2based a transposon of use makes method Our characterised bydelayedandreduceddevelopmentofmelaninpigmentation. It is practical to screen as many as 10,000 embryos for targeting, even in a small fish facility, fish small a in even suggesting thatthisapproachhasareasonablechanceofsuccess. targeting, for embryos 10,000 as many as screen to practical is It type wild the to reversion of rate the the phenotype. and genome from the excised into construct easily targeting excised and the of inducibly be insertion of can rate the measuring currently Weare females. construct and males both in genome targeting line germ the that shown have We reversion towildtypeatthegoldenlocus. Upon crossing and heatshock, unidirectional site specific recombination mediated by the y nrdcn a obe tad ra (S) I picpl te S wl iiit homologous the of initiate correction will the in DSB result the and locus principal, target the In within (DSB). recombination break DNA strand the double linearise then a will introducing I-SceI by construct. targeting the circularises and releases integrase As a model system we aim to correct the correct to aim we system model a As applied tomanylociandpossiblyevenotherteleosts. easily be can successful if and nature in generic is it establish, to consuming time is approach in described that to similar approach an using zebrafish in system recombination, homologous by targeting, gene a establish Weto aim by homologous recombinationlimitsitsutility. genome the modify to ability the of lack the but organism model important an is Zebrafish Genetics, University ofNottingham, Nottingham, UK R.H. Brookfield,F.Dafhnis-Calas andW.R.A.Brown Gene Targeting inZebrafish by HomologousRecombination Emerging technologies 2. 1. An insertion targeting construct containing an 18Kb section of the of section 18Kb an containing construct targeting insertion An An excision construct, which inducibly directs the expression of I-SceI endonuclease I-SceI of expression the directs inducibly which construct, excision An and for sites attachment by surrounded site, I-SceI an and 5 exon in nucleotide type wild the φ φ C31 integrase. C31 integraseusingaheatshockpromoter(HSP70). Drosophila Drosophila GolB 1 allele of the of allele by Rong and Golic and Rong by SLC24A5 locus which is which locus SLC24A5 Golden 398 (2000). Although this Although (2000). GolB locus with locus Golden 1 mutation and mutation φ 469 C31

Posters Posters § 470 T. Syker What atiny fishcando– From toxicityscreenstodiseasemodelwiththezebrafish embryo ec, NFS am t dvlp tcnlg pafr ta, n h oe ad sis legal suits hand, one the the otherhanddeliversaversatiletoolfordrugdiscovery. on that, platform technology on and testing, for used a authorities’animals of number the reduce to requirements industry and develop to aims UNIFISH Hence, effect additional endpoints, likemetabolomicsdatae.g.,tomeetindividualcustomer oruserneeds. including by extended be flexibly can knowledge-base The lines. transgenic in-situ hybridisation screens with wildtype knockdowns, or morpholino mutant zebrafishwith embryosexperiments orloss-of-function from studiesfrom involving e.g. obtained function, gene or relationships systematic biomarkers can easily be extracted of from the data, and follow-up studies will add information on establishment the for simulations of indispensible effect of unknown compounds. Compound-specific gene expression patterns and is which information key and zebrafish embryo benefit toxicology and great MOA-related data of for a multifactorial be vastcontain array a will knowledge-base can The companies. offood even or chemical compounds,pharmaceutical, of for knowledge-base value thus providing core a the Such form compounds. active will for library knowledge-base The library. data mode-of- toxicology compound-specific and extensive an (MOA) establishing action database, a in integrated are analysis). data transcriptome All (initially genomics things, other performed among microscopically with, combines analyses and morphometric approach comprehensive a pursues UNIFISH test. embryo zebrafish the on based assays, automated and applicable universally develop to assets disadvantages compared to e.g. the rodent zebrafish’model. The the project UNIFISH takes applications, advantage of screening these For discovery. outstanding drug track record as in a vertebrate development screening and genetic targetmodel outweighs by for far its few tool smart a also is test embryo zebrafish toxicology,the environmental and human in screen as non-target use a its Besides applications. scale-screening large to medium for potential excellent shows toxicity testing is the zebrafish embryo assay. This assay proves a very versatile platform and also the development of humans or animals. An exceptionally promising candidate for developmental alike are urged to adequately assess possible adverse the effects evaluation of of environmental and chemicals, for human context like regulatory health the in risks. pharmaceuticals, effects teratogenic Governmental of on assessment authorities the to and paid is industries attention particular reason, this For development. vertebrate in period susceptible most the is embryogenesis Early Aachen University, ofEcosystem Department Analysis, Aachen, Germany Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany; *RWTH 399 §, M.Reuter § *, V. Schiller § *, A. Wichmann § *, C.Schäfers § , M.Fenske Emerging technologies § of humandisorder. and validates the use of zebrafish GR vertebrates zebrafish of use across the validates and disorders anxiety stress-induced of machinery and symptoms the in emphasizes uniformity study this in presented result The effect. buffering social suggesting behavior, behaviors. these Finally,in mutant fishsystem that are allowed visualserotonergic interaction with other fish dobrainstem not display the freezing for role potential a toward point treatment, treatments. Accordingly, changes in serotonin transporter mRNA levels before and following SSRI behavior is inhibited by This conditions. acute similar to (Diazepam) exposures repeated and following arena isolated chronic in fish (Fluoxetine) homozygote the anxiolytic and antidepressants in basal, non-stressed state. Behavioral testing reveals significant increasecortisol inplasma freezingas behavior of well as mRNA (POMC) proopiomelanocortin and (CRH) releasing-hormone impaired negative feedback may account for the observed increase in hypothalamic corticotropin- of GR abolishes all transactivation and transrepression activity. Dysfunctional GR domain DNA-binding the in substitution R443C that reveals assay transfection transient vitro In to study the involvement of dysfunctional GR in the etiology of stress-induced anxiety disorders. susceptible to the detrimental effects of stress and thus offer an excellent model system in which denotes heritable predisposition for major depression following stressful life events. Here we Here events. life show thatadultzebrafishwithapointmutationintheglucocorticoidreceptor(GR often stressful following function, depression receptor major glucocorticoid for predisposition in heritable deficit denotes by promoted HPA, the of control feedback negative Altered disorderss. psychiatric stress-related with patients in findings neurobiological Hyperactivation of the hypothalamo-pituitary-adrenal (HPA) system is one of the most prominent theglucocorticoid in Sheba CancerResearch Center, RamatGan, Israel mutation point L. Ziv, with S.H.Meijsing,D.Strasser, A. Muto,K.R. Yamamoto andH.Baier zebrafish in receptor behavior anxiety-like Increased Emerging technologies s357 mutant for exploring the neural basis of certain aspect certain of basis neural the exploring for mutant 400 s357 s357 protein and ) aremore 471

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Hawkins zebrafishbrain.org: anonlineneuroanatomicalresource Department of Cell and Developmental Biology, Anatomy Building, UCL, London, UK;London, UCL, Building, Biology, AnatomyDevelopmental and Cell of Department 401 1 , K.Turner 1 , M.Folgueira 1 , J.D.W. Clarke 2 , S.W. Wilson Emerging technologies 1 2 MRC Investigaciones Cardiovasculares Carlos III, Carlos Cardiovasculares Investigaciones Cantabria, M.P. Cajaraville A. Barrallo Figueras A. CSIC, Cinegéticos, 17 11 1 . Pardo A M. 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Posters Posters 474 might and RGCs uncover generalmechanismsduringneuronaldifferentiationinothermodelsystems. in retraction apical underlying events molecular the on light shed will study and immunocytochemistry of combination a use we this, under are Studies way to activation. test if a similar mechanism receptor applies to apical retraction Robo during RGCs differentiation. Tothrough do N-cadherin, molecule, adhesion surface Interestingly, a previous report has demonstrated that Slit can induce downregulation of the cell so do but wildtype in as stages while stillretaininganunretractedapicalprocess. similar at axons extend RGCs morphants, these In retraction. development retinal zebrafish during that observe we that it is expressed at the apical surface of the retina. Furthermore, using time-lapse approaches, of expression the the repellent guidance molecule, Slit1b, is involved in apical process retraction. We first checked This study focuses on the molecular basis of apical process retraction. So far, we discovered that initial the While cell. underlying apicalprocessdetachmentremainsunknown. each of surface basal the from outgrowth extended of the is retinal axon axon from an the basal and process has retracted been extensively are studied, the mechanism processes Subsequently,apical membranes. retinal the to attached remain processes basal and the at generated are neurons post-mitotic apical surface of the retinal differentiation,epithelium. Their cell bodies then migrate basally, (RGC) while their apical United cell ganglion retinal Cambridge,During University, Cambridge Neuroscience, Kingdom and Development Physiology, the in GK. Wong, Neurogenesis C.Norden,L.Leung,W.A. Harris during Detachment Apical Zebrafish Retina of Understanding Mechanistic a Towards 403 mRNA during the onset of RGCs differentiation in zebrafish and found and zebrafish in differentiation RGCs of onset the during mRNA slit1b opat so dlyd apical delayed show morphants slit1b live-imaging approaches. Our approaches. live-imaging vivo in Live imaging during thematurationofunderlyingcircuit. and where correlated and anti-correlated groups appear will provide insight into the steps taken correlations between individual neurons during this developmental period. An analysis of when of development the follow sporadic to imaging calcium from time-lapse using activity,are progresses coordinated we to network the how understand Tofurther observed. is spontaneous alternation however,nor (18hpf), synchronization neither earlier other,and each hours of Twoindependent are side. motoneurons individual right in events alternation and and left synchronized the tightly between present are is motoneurons ipsilateral spinal the in in transients activity spontaneous calcium of cord, onset the after hours three (hpf), post-fertilization hours 20 At development. of stages different during patterns activity their observe to motoneurons to of Using populations the UAS:GAL4 confined system, we of are targeting activity the the genetically-encoded calcium monitor indicator GCaMPspinal cordneuronsinanintactanimalandthroughdevelopmentaltime. to techniques optogenetic apply to us allow zebrafish of transparency and accessibility genetic The development. in arise patterns activity highly-coordinated these how in Weinterested are cord. the of side right and as left the between such models alternation vertebrate and Global cord spinal in the of locomotor-likeregions ipsilateral potentials. in synchronization be with chick, and to action rat the shown of been have bursts activity this rhythmic of spontaneous, patterns display hippocampus and retina cord, spinal the as such networks system, nervous the to input provide systems sensory Before Molecular andCell Biology, UCBerkeley, USA E. Warp, C.Wyart, F. DelBene,H.Baier, E. Y. Isacoff Live ImagingofDeveloping Connectivity intheSpinalCircuit oftheZebrafish Embryo Live imaging 404 475

Posters Posters 476 visualize cellularprocessesandorganelles. expected. as works Consequently, construct it should be multicistronic possible to expand this this method with by using a live-imaging third fluorescent proteinconfocal the to these that in concluding embryos stacks, living of cells single identify to able are Wemicroscopy. by confocal embryos these analyze preliminarily to able were and embryos into construct this now, we injected to Up the proteins. fluorescent label different to with embryos able fish the are of we nuclei minitol2- and construct membranes contains this that With fishlines. vector transgenic generate multicistronic to a sites constructed integration already we aims, these reach To withthe theircorrelation and quantificationofcellularprocesseslikemigrationorcelldivisionin-vivo. automate and adouble-labeled types cell togenerate imaging softwares Image J and Volocity. distinct This method could be a basic tool for the identification and analyze wewant examine Therefore, to fishline, embryos. zebrafish living in descriptions and analyzes various cell identities in-vivo by their cytoplasmic/nucleic volume ratio and shape cellular processes. We want to develop a computer-based method that automatically recognizes with linkable be should organelles of shape and volume like parameters Basal function. cell to measure and physical track identify,parameters to of tool cells no is and there nuclei, Currently during functions. vertebrate variable development consequently allowing and to correlate them properties morphological different with types cell distinct are there In-vivo transport. molecule Cells have to carry out numerous functions that range from replication and energy conversion to Physiological I, Chemistry Biocentre, University of Wuerzburg, Germany D. Schul, ManfredSchartl,GregoryHarms,Toni Wagner In-Vivo quantificationofcellularprocesses by high-resolutionmicroscopy 405 Live imaging 1 F. Ruf-Zamojski In totoimagingofsomitogenesisandmuscleformationinFlipTrap zebrafish embryos Live imaging combining imaging of the trapped protein with in situ hybridization. Timehybridization. situ - in with protein trapped the of imaging combining by mRNAs specific the of distribution the protein compare we endogeneous and immunostaining, recapitulate using proteins expression trapped the that show We recombination. cre-lox in vivo the specific function of the trapped genes, before and after disrupting their function with assess to used is and proteins muscle endogenous to fusions protein fluorescent functional with a lines 10 identified far so has approach screening novel This mutants. conditional 2) cre-lox create and single-cell microscopy, confocal toto to and muscles in and expression protein fusion in citrine labeling with lines isolate to FlipTrapscreen cell 1) genetic individual non-invasively: arrays”, “embryo and using high-resolution imaging with degeneration and formation muscle study life. to embryos zebrafish throughout living in degenerate methods powerful and two combined maintained we end, Tothat are form, muscles high how with understand processes these to observe that resolution to species important therefore vertebrate is It in formations. skeletal mechanisms to rise conserved give highly are myogenesis and Somitogenesis Medical School, Boston, MA, USA in muscleformationandmaintenance. Together, embryos. proteins specific zebrafish by played roles the into insights living more gain to us enable approaches two in these resolution unprecedented with proteins trapped the of distribution the and formation muscle of process the watch to us allows protein fluorescent the ilg, aiona nttt o Tcnlg, aaea C, USA CA, Pasadena, Technology, of Institute California Biology, 1 , S.Megason 2 , L.Trinh 1 andS.E.Fraser 1 406 2 Systems Biology, Harvard Harvard Biology, Systems lapse microscopy of microscopy lapse 477

Posters Posters 478 how abiologicalprogramisexecutedindevelopingembryo. red with fluorescent protein. tagged Together,are theseFlipTrap studies will enable us to generate a mechanisticfusion synthesis of the of versions mutant and truncated recombinase, Cre of a generate FlipTraps lab. our tissue-specificin anddeveloped functional endogenousalso fusion withapproach yellow fluorescentmutagenesis protein. Intrap the presence gene conditional GRN, we lineage are screening RB for genes that the are expressed of in the RB components lineage using the novel FlipTrap,Todefine GRN. a lineage novel RB the of components proposed being developed in our lab. Recently,suite software a candidate gene segmentation approach had and been used to analysis characterize image source open an GoFigure2, using tree lineage RB the generate to sets data imaging our analyzing are we Then, proteins. fluorescent different photon imaging of zebrafish embryos wherein all the cell nuclei anddoing membranesare arewe tagged with First, neurons. RB of lineage cell complete and specify, induce, that (GRN) using Weare network unknown. are lineage regulatory this determine gene the and neurons RB complete of the However, lineage types. cell cell both to Thus, rise cells. give RB to and proposed was NC group” of “equivalence number an the reduce to previously shown were Olig3, and Prdm1a as such NPB, the in expressed are that factors transcription of Loss response. escape larval and Rohon- and (NC) crest neural Beard (RB) neurons, which are early-born mechanosensory neurons essential for the embryonic both to rise give to the shown ectoderm, been has non-neural (NPB), and border plate sheet neural such neuroectodermal anamniotes the other between and boundary the zebrafish frog, In the tube. as neural a form to midline the toward converge that cells neuroepithelial of emergessheet cord a spinal from the development, vivoin in Early . circuits function developmental watch to system model powerful a is cord spinal zebrafish The ofSystemsDepartment Biology, MedicalSchool, Harvard Boston, USA R. Noche andSeanMegason and Imaging FlipTrap toto Screen In using Cord Spinal Zebrafish Developing the in Lineage Neuron Sensory Identification and Functional Analysis of the Gene Regulatory Network that Regulate a Primary 407 imaging technology to elucidate the elucidate to technology imaging toto in time-lapse confocal and two- and confocal time-lapse vivo in Live imaging and Biophysics Unit, EMBL, Heidelberg, Germany; Heidelberg, EMBL, Unit, Biophysics and Tae ae novd n euaig h drcinl irto o meod el rsodn to responding cells myeloid of migration directional inflammatory cuesinvivo. the regulating in involved are GTPases pharmacological and Kinase inhibitor. inhibitor Rho elongation Collectively,a of cell our presence data support the abnormal the in pseudosubstrate hypothesis occurs that as the speed Par/aPKC cell complex asymmetry and in its myr-PKCreduction effector macrophage ξ concomitant stabilize to a well as with contributing is signaling signaling RhoA cells. untreated with compared when PKCξ motility directional macrophage impairs blocking Accordingly, cells. reveals morphologies cell increased number of of unpolarized pseudopodes and elongated phenotypes in mutant-expressing inspection Closer injury. the to response delayed a mutant- outcome, an orl_Par3 cells, as and, status control non-polarized with time” with “sensing prolonged a Compared display macrophages expressing binding. Par3/aPKC with interfere to known approach, mutant deletion dominant-negative orl_Par3 transgenesis the macrophages in specifically overexpressed we transient a using By migration. directed and polarization macrophage complex signaling this of role functional the explore to system polarization in several cell types. We used a Medaka latipes) (Oryzias transgenic line as a model PKC (aPKC) in conjunction migratory with leukocyte small for GTPases requirement of a the response is Rho to Family polarity inflammatorytemporally cell cues.and of spatially Amaintenance controls conservedand polarityestablishment complexThe consisting of Par3, Par6 and atypical Heidelberg,Germany 1 1 The Par/aPKC ComplexControls The Vectorial Migration OfMedakaMacrophagesIn Vivo Cell movement Leukocyte Biology Unit, S. Raffaele University School of Medicine, Milan, Italy; Milan, Medicine, of School University Raffaele S.Unit, Biology Leukocyte C. L.Crespo, 1 R. Molteni, 1 B. Clissi, 2 P.Keller, 3 J. Wittbrodt and 3 Heid

elberg Institute for Zoology-Univ.of for Institute elberg 1 during wound-triggered during vivo in R. Pardi 408 2 Cell Biology Cell 479

Posters Posters 480 plate cellmovementsandorganise theirpolarity. are We process. this in role neural important of co-ordination the regulate can together mesoderm and ECM whether testing currently an have to may tissues embryonic two these between expressed are which (ECM), matrix extracellular the of components tissue-interaction, such of mesoderm- in rescued nature molecular the know be not do we Although, mesoderm. can replacing by embryos mutant morphogenesis less tube demonstrate neural we addition, and In organisation development. apico-basal tube that neural for required is itself signalling that Nodal rather mesoderm of lack that suggest experiments our morphology, tube neural defective are severely disrupted in these mesoderm-less embryos. Although, Nodal mutant embryos show Time-neurulation of organisation.movements co-ordinated highly normal apico-basal the that demonstrates microscopy chaotic lapse with primordium neural a develop signalling, Nodal is convergence of loss to due plate mesendoderm head lack completely that neural embryos Furthermore affected. severely that find we function, Has2 of reduction mesoderm the defective to with due embryos convergence in example, For embryos. mutant mesoderm of series a in mesoderm move in a tightly co-ordinated way. In addition time-lapse microscopy we have are revealed analysing that neural during the morphogenesis initial stages of neurulation, might neural actively contribute to plate co-ordination of this and process is the adjacent mesoderm. Analyses by morphogenetic coordinated ahighly the dorsal midline and the generation of the apico-basal cell polarity.is One of the key tissues that towards cells neuroepithelium neuroepithelial of convergence the involves This space. and time both in zebrafish process the of development The and polarization organising in MRC Centre forDevelopmental Neurobiology, KCL, matrix London, UnitedKingdom extracellular C. Araya andJ.Clarke and mesoderm morphogenesis ofzebrafish neuroepithelium of role Potential 409 Cell movement modification on to proteins with the CAAX motif including the core PCP protein Prickle (Pk), Prickle protein PCP FT,of abrogation core GGT,not but the the enhances including motif CAAX the with proteins to on modification lipid performs preferentially FT that notion the with Consistent backgrounds. mutant/morphant farnesyl transferase (FT) or geranylgeranyl transferase (GGT) function using morpholinos on PCP the possibility that the prenylation pathway test regulates proteins. To of activities C-terminus of the at the modification PCPlipid for pathway,implicated been has we and HMGCR, abrogated other core PCP mutants/morphants. The prenylation pathway is one of branches downstream of extension at pharyngula stages. Notably, abrogation of yolk thicker a showing while gastrulation during phenotype CE mild very only exhibit embryos the of 1b(hmgcr1b) enhancer dominant a Areductase as gene 3-hydroxy-3-methyglutaryl-Coenzyme identified have we screen, is and regulated co-ordinated by a of non-canonical variety Wnt/planar a cell polarity by (PCP) established pathway.directed movements is of cells. axis From One body our of these forward the movements genetic is gastrulation, convergence and vertebrate extension (CE), During which Institute ofMolecularCell BiologyandGenetics, Dresden, Germany 1 M. Tada Regulation ofplanarcellpolaritysignallingby theprenylation pathway Cell movement the specificityfortargets oftheprenylationenzymes. Department of Cell and Developmental Biology, University College London,UK;Biology,College DevelopmentalUniversity and Cell of Department 1 , M.Kai 1 , N.Buchan 1 , andC-P. Heisenberg E phenotype. CE (slb)/wnt11 silberblick or pk1a 2 morphant CE phenotype, suggesting phenotype, CE morphant pk1b hmgcr1b also enhances the CE defects of 410 mutant hmgcr1b 2 Max Planck Max 481

Posters Posters 482 analyses, weaimtogeneratea4Datlasofnormaleyedevelopmentinzebrafish. comprehensive these Through segmentation. eye using lens and retina the on topographically counting software, and manually identify mitotic figures to generate a 4D map of mitoses, plotted morphogenesis. We measure cell numbers from the histone-mCherry data using custom 3D cell We have also begun to analyze the relative contributions of cell division morphology andcellmovementswithinthisdevelopingtissue. and cell migration to eye combined with the eye segmentation as polygon meshes, allowing us to visualize changing eye be then can cells individual Tracksof cells. tracked the of images export volume can or allows which coordinates and XYZ application time, LongTrackthrough backwards The or forwards either tracking. cells individual cell track to 4D us manual datasets. for 4D software these custom for developed developed We Amira. in have segmentation manual using we time over morphology that retina and lens methods analyzed We analysis several describe we Here with histone-mCherryandEGFP-CAAX,respectively. 24 the eye morphogenesis, we zebrafish carry out obscuring live 4D imaging of the of developing eye primordium from 12 to thus study our and during problem animal, this Tocircumvent same morphogenesis. the of aspects of dynamic development the analysis during preventing points time fixation, different require of methods these However, morphology. their visualize SEM to or sectioning used commonly have biologists tissues, of development the Tounderstand University ofUtah, Dept. & Neurobiology Anatomy, SaltLake City, USA H. Otsuna,K.Kwan,C-B.Chien 4D timelapseanalysisofzebrafish eye development hpf, collecting complete z-stacks every 3.5 min after labeling chromatin and cell membranes 411 Cell movement duckling ( in analysis expression Gene udu expected The normal. largely is embryos the shortened tail than and either Interestingly, malformation single mutant. heart death. Furthermore, head, cell smaller tail,increased degenerated axis, body shortened show embryos uain ht eeae a rnae poen Moreover, protein. truncated a generates RT-PCR that sequenced mutation We (2007). al. et from amplified Liu gene udu by of mutant products defect subsequently erythroid and primitive 1996) a al., as et reported (Hafter morphogenesis affecting mutation a as screen genetic enhancer of the 1 A. Sawada and extensionmovements Ugly duckling, SANT domain protein interacts with Wnt/PCP pathway to regulate convergence Cell movement iceity n Bohsc, CF Sn rnic, USA, Francisco, San UCSF, Biophysics, and Biochemistry To identify additional genes involved in C&E movements, we performed a genetic screen for screen the genetic modify that a mutations performed we movements, C&E in involved genes additional Toidentify molecular mechanismsthatcontrolC&Emovementarenotfullyunderstood. as such pathway PCP Wnt/ of components in mutations zebrafish, In movements. (C&E) extension and convergence various undergo regulates and pathway Wnt/PCP extension. internalization and epiboly,convergence as by such movements form of types layers germ the gastrulation, vertebrate During hypothesize thatuduregulatesC&Emovementsactinglargely inparalleltoWnt/PCP signaling. that Wnt/PCP pathway genes are normally the expressed in suggest analyses analyze expression Moreover,gene defects. C&E Toenhanced shows mutant double The defect. and C&E MZudu generate also including we pathway,Wnt/PCP with Udu abnormalities of interaction morphogenetic severe show embryos generated we development, during maternal-zygotic (MZ) Udu of function the by determine Tocompensated products. is maternal function the zygotic Udu that suggesting 2007) al., et (Liu gastrulation during 2007). al., et (Liu protein the Udu for of essential function is domain this and region domains C-terminal the conserved in some domain SANT contains putative It including function. unknown largely of protein acid amino 2,055 University ofGuelph, Canada, eatet f ilgcl cecs Vnebl Uiest, ahil, USA, Nashville, University, Vanderbilt Sciences, Biological of Department sg1 mutant allele. Thus we conclude that conclude we Thus allele. mutant ) mutant locus in chromosome 16. chromosome in locus mutant udu) 1 , C. Yin kny 2 , T. Van Raay mutant phenotype. knypek/glypican4and udu mutant by using germ-line replacement (Ciruna et al., 2002). MZudu phenotype. We identified We phenotype. (kny) knypek vu66;knyof patterning general the that suggests mutant compound 4 School ofBiologicalScience, University ofLiverpool, UK 3 , T. Wilm obe uat mro ehbt oe severely more exhibit embryos mutant double vu66;kny homozygous embryos and discovered a nonsense a discovered and embryos homozygous vu66 vu66 is a recessive lethal mutation and the homozygous 4 andL.Solnica-Krezel is expressed maternally and then ubiquitously then and maternally expressed is udu trilobite/vangl2 vu66 is a new allele of allele new a is vu66 was initially identified in the Tubingen the in identified initially was udu mutation point is located very close to close very located is point mutation vu66 udu mutant embryos. Taken together, we also interacts genetically with vu66 cause a C&E defect. However,defect. the C&E a cause 3 oeua ad ellr Biology, Cellular and Molecular uat os o complement not does mutant 1 412 mutant as a recessive a as mutant vu66 udu locus. double mutant. double kny 2 eatet of Department encodes udu trilobite. 483 ugly

Posters Posters 484 and wewillreporttheirinteractionswiththetwosystems,CXCR4/CXCR7WNT/FGF. We have identified yet other components ofLecaudey etal,2008, Aman andPiotrowski,2009). the network that regulates migration of thethat the WNT/FGF system also plays a role in driving migration ( Nechiporuk and Raible, 2008, primordium the primordiumandthatCXCR7providesdirectionality.of Recentworkbyothershaveshown migration the for required is CXCR4 that suggest results Our cells). leading the in all at not and primordium, receptors two its by and migration, CXCR7. and of CXCR4 path the along present is which SDF1, the chemokine at the cells of Groups tail. by driven is the primordium the of migration the organs.Wethat sense shown as have differentiate of to settle tip eventually and time the of intervals regular to at down slow moves primordium the of and edge trailing vesicle otic the near originates that primordium a of migration the on depends teleosts of line lateral posterior the of formation The University ofMontpellier2, Montpellier, France. C. Dambly-Chaudière,L.Gamba,G.Lutfalla,and A. Ghysen Control ofcellmigration inthedevelopment ofthelateral line 413 is expressed in the reciprocal pattern (highly in the trailing cells and cells trailing the in (highly pattern reciprocal the in expressed is cxcr7 is expressed in the leading cells but not in the trailing cells of the of cells trailing the in not but cells leading the in expressed is cxcr4b Cell movement Timely generation of distinct neural cell types in appropriate numbers is fundamental for the for fundamental vertebrates In is retina. functional a of generation numbers appropriate in types cell neural distinct of generation Timely Instituto Cajal, CSICandCIBERdeEnfermedadesRaras(CIBERER), Madrid, Spain R. Marco-Ferreres, I.Conte,L.Beccari,E.Cisneros,J.M.Ruiz,N.Tabanera andP. Bovolenta NeuroD andSix6 between loop regulatory feedback a on depends photoreceptors rod of Timelydifferentiation Cell movement Supported by:Supported Six6 feedbackregulatoryloop. NeuroD- a requires differentiation subsequent their and precursors photoreceptor of generation of regulator direct a induces Six6 is NeuroD of that indicate results function Togetherthese marker,rhodopsin. differentiaion loss-of photoreceptor the of localization and and expression the gain- in alterations and Interestingly levels expression NeuroD types. in changes cell retinal other in differentiates, changes retina the as Notably,apparent no with differentiation, photoreceptor studies. over-activationimpairs Six6 NeuroD-mediated gain-of-function Six6 previous in proliferation, with progenitor agreement retinal unorganized promotes embryos fish medaka in over- expression Six6 NeuroD-induced retina. the in expression gain-of-function Six6 regulating enhancer, with this in present together assays ChIP and factor,transcription bHLH a sequence NeuroD, “E-box” that an indicated binds studies directly luciferase sites, binding factor transcription the transgenic in and searches bioinformatic used have we approaches in medaka latipes), fishidentifying (Oryzias a highly conserved regulatory enhancer issue this address to begin Tounclear. How zone. to restricted marginal ciliary proliferative becomes the to as well as then cells amacrine and ganglion retinal differentiated and progenitors retina multipotent in expressed initially is family, ou rsosbe o is xrsin n ifrnitn ad dl rtn. erh for Search retina. adult and differentiating in expression its for responsible locus Six6 expression in the retina is controlled and what are its precise functions is still is functions precise its are what and controlled is retina the in expression Six6 BFU2007-6177andCAM, P-SAL-0190-2006 expression in the eye development and suggest that the appropriate the that suggest and development eye the in expression Six6 , a transcription factor of the of factor transcription a Six6, 414 Six/sine oculis Six/sine 485

Posters Posters 486 two contains genome latipes) (Oryzias member the a Six3, of including factors transcription several of expression overlapping the by plate Vertebrate forebrain derivatives are specified at early stages of gastrulation in the anterior neural Instituto Cajal, CSICandCIBERdeEnfermedadesRaras(CIBERER), Madrid, Spain L. Beccari,I.Conte,E.Cisneros,N.Tabaneras andP. Bovolenta olSox2-mediated activation ofolSix3.2promotestelencephalicversus eye fieldspecification Supported by:Supported is mostlyresponsiblefortelencephalicspecification. fish, medaka in that with interference that associated with an expansion of the telencephalic territory. Because previous studies have shown embryos, up-regulates of expression late and early forebrain and its derivates. Consistent forwith these results, over-expression of Sox2 in medaka responsible fish enhancers the activate and bind directly Pax6 and Sox2 that determined further have we assays, interaction chromatin-protein basis of searches for transcription factor the binding On sites, 8(7):R137). 2007; expression Biol. analysis, Genome luciferase Bovolenta, and reporter (Conte and blockers silencer and silencers proper for necessary code possibility genes and decipher this the two test logic to Toof begin the duplication. genome after of evolution independent sub-specialization their of consequence functional as and regulatory possible a suggesting regions, preponderant expression of avian or mammalian the of that recapitulates genes two these of pattern expression 415 cls aiy f oebx otiig rncito fcos Te eaa fish medaka The factors. transcription containing homeobox of family oculis Six/sine BFU2007-6177andCAM, P-SAL-0190-2006 functions have been segregated between two paralogs, where paralogs, two between segregated been have functions Six3 expression affects predominantly eye development, we propose we development, eye predominantly affects expression olSix3.1 olSix3.2 levels. Notably, this up-regulation causes severe microphthalmia olSix3.1 in the eye and of olSix3.2 Six gene regulation, we have identified the precise regulatory xrsin Ti cd cmrss sre o enhancers, of series a comprises code This expression. Six3 paralogs: olSix3.2 in the telencephalic and thalamic and olSix3.1 Te combined The olSix3.2. Cell movement n the in olSix3.2 , with a with Six3, olSix3.2 myod/myf5 in thezebrafish. that the early myotome is essential for subsequent skeletal muscle differentiation and patterning show also data our Therefore complete. not was inhibition morpholino the abnormally,when albeit proceed, does myogenesis whereas abolished, is myogenesis subsequent myotome, a of absence complete the in that Wereport myotome. a of absence the in occurs myogenesis how Moreover, because of the transient nature of morpholino inhibition, we were able to investigate differentiation. muscle of onset the at only expressed is embryo, mouse the in that unlike gene, the because spontaneously, happen not does Rescue vitro. in function chromatin-remodelling 1 muscle development. Here we show that show we Here development. muscle and sufficient to activate (albeit not completely) skeletal myogenesis. In the zebrafish embryo, in great detail only in the mouse embryo, where all but myogenin – Myod, Myf5 and Mrf4 – are studied been has role their but vertebrates, all in myogenesis activate factors regulatory Muscle Studi diMilano, Italy A. Pistocchi myod/myf5 the in myogenesis rescues myogenin double morphantzebrafish embryo not but mrf4 of expression early Induced Segmentation Division of Regenerative Medicine, San Raffaele Scientific Institute, Milan, Italy; myf5 are required for induction of myogenesis because their simultaneous ablation prevents 1,2 double morphant via a selective and robust activation of activation robust and selective a via morphant double , E.Schnapp 2 , G.Messina 1,2 , E.Foglia mrf4 mrf4 but not but 2 , C.LoraLamia myog can fully rescue myogenesis in the in myogenesis rescue fully can 416 2 , F. Cotelli myod , in keeping with its with keeping in , 2 andG.Cossu 2 Università degli myod 487 1,2

Posters Posters Barruso and Junta deCastillayLeón,Barruso andJunta GrupodeExcelencia deCastillayLeón. Supported by grants from the Fundación Mutua Médica Madrileña, Fundación Samuel Solórzano to NIHguidelines. for the use care of animals in research (RD the 1201/2005, of BOE European guidelines 252/34367-91, Communities Directives (86/609/EEC the 2005) and 2003/65/EC), with and the current accordance Spanish conformed legislation in were protocols experimental and procedures the All the disrupt may may affectthesignallingpathwaysrequiredforthisphenomenon. ethanol development, embryonic evagination of the optic vesicles. The alteration in the expression of visual field early patterning genes the produce that shape cellular during in changes key the impair and field eye the that, of cytoarchitecture indicate results Our after theethanoltreatment. in holoprosencephaly and cyclopia, we have observed a reduction in the expression of this gene treatment. ethanol after preserved is it vesicles, optic between area the in present not is expression rx1 somites-stage, 10 at animals, control in while Moreover,vesicles. optic the of evagination incorrect an indicating ethanol, to exposition after the ethanol, not evident and several pyknotic to nuclei can are be observed. morphology exposition The cellular expression pattern in of changes the animals, control in than After wider is cells somites-stage). between separation (10 fusiform to somites-stage) (3 circular 10-somites at stage and two optic vesicles are stage, observed. The cellular morphology 3-somites in control animals changes the from from distinguishable is field eye the animals control In evagination), 6somites(mid-evagination)and10(post-evagination). to (prior somites 3 are analyzed stages The group. experimental as tailbud to stage dome from solution ethylic 2.4% to exposed embryos and control as embryos untreated zebrafish used AB 488 as such formation, eye for required genes of patterns sectionsand expression eye semithin using the from thesedomains derived of precursors against retinal immunohistochemistry the cytoarchitecture the of analyzed characterization have the We is field. work this of aim The largely are unclear.vesicles optic the of morphogenesis the affects ethanol which by mechanisms The vesicles, resulting in cyclopia, a condition where the optic vesicles are partially or totally fused. Within the anterior neural plate, ethanol has been reported to avoid the evagination of the optic organization. cellular disrupts and system nervous central the of ventralization impairs Ethanol Cell BiologyandPathology, INCyL, Salamanca, Spain A. Santos-Ledo,F. Cavodeassi,R.Sánchez-González,M.Moyano, A. Porteros,J. Aijón, R. Arévalo Ethanol alterstheestablishmentofeye fieldduringdevelopment 417 β-catenin. hybridization was carried out to determine the determine to out carried was hybridization situ In missense mutations has been reported been has mutations Six3missense , rx1 and rx3 Segmentation six3. rx3 is altered e have We ß2. through gadd45 However, signals. Our data suggest that the downregulated strictly signalling tobeapotentialtargetnotochord, indicatingdro1/cl2 oftheHhpathway. Hh of inhibition the converse, This knocking-down effect. specificity ofthedro1/cl2 in mRNA ß2 gadd45 of injection The somites. formed already the in disappeared expression its unchanged, almost Interestingly, while the expression pattern of marker clock her1, segmentation and The the one. segment adaxial polarization the markers in unaltered were while mesoderm, paraxial markers myogenic The somitogenesis. of phases different during expressed markers of alterations possible investigated we Therefore, overexpressed embryoswerenotaffected. the while somitogenesis, early the in somites shaped abnormally showed Morphants specific a injecting by experiments Tothe of role functional the investigate both of muscles the in trunkandhead. detectable becomes it stages developmental later In notochord. the to E. Villa 1 I. Della Noce the of somitogenesis characterization functional and analysis Expression Segmentation epiboly. Interestingly, during somitogenesis the somitogenesis epiboly.Interestingly,during Whole somitogenesis. late to up segmentation mount of steps initial the from increasing starts level The spatio-temporal analyses evidence a maternal and zygotic nuclear multiple and localization signals. thepeptide signal presents a of aa) presence (867 contemporary the protein showing translated structure, same the and and counterpart homolog mammalian the zebrafish with its homology cloned we DRO1/CL2, The development. embryonic during of function its analysed role physiological the understand To is and tissues adult oncosuppressor gene. all almost in expressed is downregulated in gene several cancer cell lines and DRO1/CL2 solid tumors. DRO1/CL2 is indeed human considered an cloned recently The Milano, Italy; nvrià el Sui i oea Rgi Eii, oea Italy; Modena, Emilia, Reggio e Modena di Studi degli Università dro1/cl2 dro1/cl2 1 in situ hybridization experiments show a diffuse , F. Cotelli 1 , A. Pistocchi 3 loss-of-function effect on the somites could be due to a disruption of nothocordal of disruption a to due be could somites the on effect loss-of-function Stazione ZoologicaStazione “A. Dohrn”, Naples, Italy 2 , F. Schepis and tail no MO injected embryos rescued their phenotype, confirming the confirming phenotype, their rescued embryos injected MO dro1/cl2 dro1/cl2 notochordal signal may play a role in lateral somitic maturation 2 , E. Turola 1 ee nfetd in unaffected were hedgehog sonic 1 , C. Brusegan O n te ul egh RA respectively. mRNA, length full the and MO dro1/cl2 dro1/cl2 dro1/cl2 and myod mesp-a and ß2 gadd45 in the anterior presomitic mesoderm was dro1/cl2 dro1/cl2 2 , S. Carra gene, eutd evl dsutd n the in disrupted heavily resulted myogenin mesp-b dro1/cl2 dro1/cl2 we performed loss- and gain-of-function and loss- performed we 2 expression becomes mainly restricted mainly becomes expression , R. Critelli zebrafish gene shares high shares gene zebrafish dro1/cl2 dro1/cl2 dro1/cl2 showed no significant alterations. ee uig zebrafish during gene dro1/cl2 expression during cleavage and 418 1 xrsin n the in expression dro1/cl2 , C. De Lorenzo 2 nvrià el Sui di Studi degli Università expression. Its expression opat. By morphants. dro1/cl2 1 , P. Sordino dro1/cl2 dro1/cl2 489 3 ,

Posters Posters 490 her1, in the form intrinsic of signaling gradients in the PSM as well as functional cyclic expressions of PSM genes, such as principles: important oscillators two at the tissue level. The existence on of a wavefront and clock relies in zebrafish was discovered PSM oscillators the in individual in cells this and clock communication tightlyregulates between cells segmentation model”, that allows the for synchronization andwavefront of the “clock in specifically,More somites. new of positioning and timing the for accounts and system complex described first was which clock, segmentation A tissue. (PSM) mesoderm presomitic unsegmented the from intervals regular at and sequentially formed are somites called structures distinct embryogenesis zebrafish During MPI-CBG, Dresden, Germany A. Oswald, S. Ares, A. C.Oates Constructing asyntheticsegmentationclock inyeast of somitogenisisinzebrafish. clock components within the segmentation clock network, thereby furthering our understanding synthetic oscillator principles, this method can elucidate the function the and dynamics rebuilding of individual by implemented be segmentation clock piece is by piece in will theseoscillators approach of synthetic a Therefore, mechanism understood. molecular well theexact not thus and embryo zebrafish the within of role component the each of function suggest the verify to models difficult is However,it components. interaction clock individual molecular with combination in phenotypes mutant of data experimental stage, this At formation. boundary proper of failure or size segment altered 419 and her7 . Mutations in these genes often result in abnormal segmentation patterns, segmentation abnormal in result often genes these in Mutations delC. S. cerevisiae (budding yeast). In conjunction with de novo Segmentation by Chambon et al. In the present study, over-expression constructs of the Notch intracellular Notch FP:ER the to fused ICD Delta or of (NICD) domain constructs over-expressionstudy, present the In al. et Chambon by human estrogen receptor (ER receptor estrogen human inducible hormonal a establish system, which can has with rapid kinetics after activation and tunable receptors induction level. fusion A mutated steroid HBD of Heterologous of signaling. (HBD) Notch domain stimulate hormone-binding the can that chemical agonistic no is there DAPT,a Notch2 and DeltaC/DeltaD) function redundantly in PSM or not. Blocking Notch signaling with Furthermore, there are still some controversies whether the Delta-Notch paralogs (e.g. Notch1a/ in particular regulation or the coordination mechanism of the segmentation clock signaling. However,are thedetailedrolesofparticularDelta-Notchcombinationsthatareinvolved still unclear. ( related downstream targets cells PSM across interaction Delta-Notch that proposed was it zebrafish, In genes. non-cyclic and patterns) expression oscillating (with genes cyclic including components core of consists which (PSM), mesoderm presomitic the in clock Repeated compartments are formed during vertebrate somitogenesis driven by the segmentation Max PlanckInstituteofMolecularCell BiologyandGenetics, Dresden, Germany B-K. Liao Conditional activation ofNotch signalingduringzebrafish somitogenesis Segmentation could beelucidated. phenotypes, the somite specific and functions effects signal of subsequent particular the comparing Delta-Notch and analyzing genes By during background. deficiency zebrafish somitogenesis her) family genes, synchronizing oscillation and causing a feedback-loop to Delta-Notch 1 -secretase inhibitor, can achieve temporal control of Notch signaling. Nevertheless, signaling. Notch of control temporal achieve can inhibitor, γ-secretase and A. Oates 1 T ), which binds to tamoxifen but not estrogen, was first developed first was estrogen, not but tamoxifen to binds which ), T will be introduced into the corresponding genetic corresponding the into introduced be will 420 and hairy enhancer of split- of enhancer 491

Posters Posters 492 and disease. model for understanding Wnt gradient formation and signaling a range in vertebrate development as serve will system developmental this from learnt lessons The embryo. zebrafish living the in formation gradient in proteins Wnt of modifications lipid of functions the determine will we secreted Wnt proteins are found in multi-component whether complexes determine in will we the Next, zebrafish embryos. embryo. zebrafish Finally, in modified lipid are molecules signaling modified, but the role of lipids in Wnt signaling is not clear.formation. Unusually for secreted signaling molecules, some Thus,Wnts have been shown to be lipid we will determine whether Wnt presomitic mesoderm and will analyse candidate cellular receptors contributing to Wnt gradient zebrafish the in distribution Wnt determine embryo. first zebrafish will the We of tailbud the in secreted completely unknown. Candidate Wnt glycoproteins proteins are Wnt3l is Wnt and tissue this Wnt8, in which formation of are gradient locally and expressed movement Wnt gradients of mechanism the but extra-cellular tailbud, the from with clock molecular a of interaction the of skin and muscle bone, axial segmented somites, adult. The the periodic formation of somites from the into to presomitic mesoderm is thought to result from rise subdivided give sequentially that is cells bodyof Somitogenesis clusters embryo system. epithelial vertebrate amodel the which as in process somitogenesis the zebrafish is using formation mechanisms understand gradient to aim Wnt we of project build this molecules In these understood. how well but not synthesis, is of gradients site signaling their from distance a at signal usually Wnts normal development, as during well proliferation as and formation regulating pattern in stem processes many cells control molecules and signaling Wnt cancer progression during pathogenesis. MPI-CBG; Dresden, Germany C. Eugsterand A. C.Oates in formation gradient study vertebrate development to anddisease system model a as somitogenesis zebrafish in signaling WNT 421 Segmentation of theoscillatingPSMcells. synchronization the affects this how and movements, cell of dynamics the understand to order in models motility cell quantitative derive to like would we data, this from extracted properties movement cell on Based embryo. zebrafish single a of cells the of nuclei the all of movements displacement, and correlation functions. Recently, Keller quantitatively characterize and the oscillations cells cellular movements mesoderm based pre-somitic the on of velocity,trajectories patternsthe acceleration, obtain mean square to the like would we and work, this In properties form. synchronization their affect oscillators of motion of role the particular, in relative of system, characteristics the that studies theoretical the from known well is of It movements. cellular properties the of measurement quantitative a have to examine the relationship between temporal program and spatial pattern, there is a general need and Toexplain achieved. from far ofthe is PSM the in oscillations understanding genetic the complete of behavior several identified, collective been Although have expression. clock the gene segmentation translating static this cycle, of of components oscillation patterns the are spatial of striped oscillations phases into These different periodicity temporal at signaling. wavefront Notch locally anterior whose an intercellular oscillators at by cellular arrested controlled of assembly is an behavior by controlled coordinated is process This every about minutes. formed is the somites 25 of embryo, pair the new one of zebrafish, tissuethe In unsegmented (PSM). mesoderm posterior presomitic the from grows embryo the the of as muscle sequentially the of and vertebrae form the the to in rise appear anterior-to-posteriorsomites an where in sequence, occurs Segmentation tail. mesoderm and trunk give paraxial that structures the repeated in are evident which is somites, embryo vertebrate the in Segmentation Institute forthePhysics ofComplex Systems, Dresden, Germany 1 B. Rajasekaran Analysis ofcellmovements inthepresomiticmesodermofzebrafish embryo Segmentation Max Planck Institute of Molecular Cell Biology and Genetics,Dresden,Germany;and Biology Cell Molecular of Institute Planck Max 1, 2 , S. Ares 2 , L.Morelli 1 , F. Jülicher 2 , A. COates et. al. published digital datasets for the 1

422 2 Max Planck Max 493

Posters Posters 494 expression atthelevelofnode. in the left-right patterning is not conserved during mouse development possibly to the loss of its on these embryonic processes. Strikingly, our results seem to indicate that the role of left-right on effect the patterning and segmentation clock synchronization we are investigating the role of Terra/Dmrt2about known is nothing since Nevertheless, differentiation. somite late that described been already has It PSM. the in symmetry left-right promote to necessary also is it but LPM the in asymmetries left-right that shown have We of embryosreliesonthesynchronizedsegmentationpresomiticmesoderm(PSM). bilateral symmetric position a maintain between must both way,somites correct sides a of in the form embryonic muscles skeletal axis. and This skeleton symmetric axial patterning the that so lateral plate mesoderm (LPM), a territory that will contribute to several organs. At the same time, internal the with is responsible for the transfer of information from regions scenario adjacent to the embryonic node to the different a see cascade genetic and complex a development, we in Early distribution. left asymmetric an acquiring organs inside the looking between symmetric when is Nevertheless, axis sides. body right vertebrate the outside, the from Desenvolvimento, looking When do Biologia e Histologia de Instituto e Faculdade deMedicina de Lisboa, Portugal; Molecular InstitutoGulbenkiandeCiência, Oeiras, Portugal Medicina de Instituto R. Lourenço,S.LopesandL.Saúde Role ofterra invertebrates 423 , a zinc finger-like transcription factor, is required tocreate isrequired factor, transcription finger-like zinc a , terra/dmrt2 terra/dmrt2 terra/dmrt2 knockout in the mouse leads to defects in defects to leads mouse the in knockout Segmentation terra/dmrt2 We willpresentoureffortstotacklesomeofthesequestionsquantitatively. cycles. of number appropriate the for going clock the keeps that oscillator,mechanism the and her1, concentrations of Her1, Her7 and NotchICD proteins, the of cross-regulatory interactions between initiation of rate the of dependence functional the delays, time the of features oscillate in the PSM, and their relative importance remains to be clarified. Many other important Other clock. the of pacemaker ultimate the is really proposed, have we that way simple a that proven from far is qualitatively.it and But quantitatively model the supported have system the of parameters key several of Measurements in synchrony, thankstooscillatingexpressionoftheNotchligandDeltaC,drivenbyHer1/7. Notch signalling between adjacent cells is then proposed to keep their individual clocks ticking to rise give conditions, certain oscillations withineachcellofthePSM. on dependent can, products protein own their by genes these of expression the of autoinhibition direct that demonstrates analysis Mathematical PSM. the in and genes, target Notch of pair a be to proposed are zebrafish in components clock core The as shows a segmentation components, “clock”, which pathway leaves its Notch trace in for the periodic coding spatial oscillating pattern mainly expression in the of presomitic mesoderm (PSM) at formed genes, the tail end somites. of the embryo, behaving of set a somitogenesis During U.K. London, Institute, Research London UK Research Cancer Laboratory, Development Vertebrate about the questions M. Holder, A. Hanisch,andJ.Lewis unanswered clock: feedback segmentation loop zebrafish the Deciphering Segmentation . Both code for inhibitory bHLH transcription factors and show oscillating expression oscillating show and factors transcription bHLH inhibitory for code Both her7. n te te oscillatory other the and her7 control loop also remain to be analysed, including the nature of the key the of nature the including analysed, be to remain also loop control her1/7 ee, h sucs f os i te gene-expression the in noise of sources the genes, her feedback loop, operating in the in operating loop, feedback her1/7 424 transcription on the on transcription her1/7 genes also genes her her1/7 her1 495

Posters Posters 496 themselves restrictedtoformingsubsetsofexcitatorylineages. cells. This suggests that the inhibitory fates are determined sequentially upon lineages, which are on in different excitatory lineages arising from Vsx2 (bipolar cell) and Ath5 (ganglion Using cell) positive GAL4/UAS system. neuron glutamatergic all of expense the the using expression Ptf1a drive we which in studies, overexpression our in shown as types at cell fates inhibitory the drive to sufficient also is Ptf1a and fates cell early the into transdifferentiate knockdown Ptf1a with cells early i.e. cell types neuron of the retina (photoreceptors, glutamatergicbipolar, the ganglion cells) of in the expected histogenic order,all into transdifferentiate cells which in down, knock by revealed as fates these determine to necessary is Ptf1a differentiate. to layers specific their to move cells Time-lapse imaging reveals that Ptf1a expression commences primarily in postmitotic cells system, whichgiveinsightsintosubtypespecificsynapticconnectivityandvisualpathways. before a Using and horizontal retina. zebrafish the in cells amacrine differentiating expressing Ptf1a of scheme classification immunohistochemical the in expressed including is system, Ptf1a nervous that amacrine cells – the show two the inhibitory neuronal types we of the retina. of We line provide a morphometric ptf1a:GFP areas and transgenic different throughout fates inhibitory of determination cell the in implicated was (Ptf1a) 1a factor transcription pancreas The the in neurogenesis zebrafish retina,anaccessibleandhighlyorganised partoftheCNS. during determination fate cell of of control mechanisms the study Wevia expression. co-ordinated gene highly is process This cells. progenitor multipotent from form During neurogenesis of the central nervous system (CNS), a multitude of different neuronal types ofPhysiology,Department Development andNeuroscience, University of Cambridge, UK PR. Jusuf andW. A. Harris excitatory all of expense cell fates the at retina zebrafish the in fates cell inhibitory determines Ptf1a 425 timelapse imaging, we can show that Ptf1a in the wild type condition is switched is condition type wild the in Ptf1a that show can we imaging, vivotimelapse in Gene regulation vice versa . vice experiment of the Mauthner neuron in the or (ngn1) both in genes: Drosophila proneural to vertebrates. of These two families families have divergent two activities in The the formation of factors. neurons transcription bHLH for code neurons of formation the to necessary are which genes Proneural Centre deBiologieduDéveloppement UMR5547, Université Paul Sabatier, Toulouse III, France R. Madelaine*,P. Blader Studying theactivity oftheproneural genesngn1andascl1ainzebrafish Gene regulation The study of this module will lead us to gain further insight in the molecular basis explaining basis molecular the divergences ofactivitybetweenproneuralgenestheachaete-scuteandatonalfamilies. in insight further gain to us lead will module this of study The byNgn1. this E-BoxandthattheyarenecessarytoallowtheregulationofdeltaA of regulation the allow to manner by deltaA Ngn1. Moreover,cooperative EMSAa experiment and transgenic lines show that Ngn1 can bind on in act to appear which (E-Box) sites binding Ngn1 potential three of cluster a is there fragment, this In Ngn1. by regulation a for necessary the of analysis functional The that suggest experiment function of loss and gain Both have we of divergences, study these a understand begun to attempt an In genes. targets different of regulation likely that divergent activity between Ascl1a differ in their abilities to save mutants. Proneural genes are transcription factor, making it and achaete-scute Drosophila and mouse. The transgenic lines, allowing the misexpression of achaete-scute-like1a ( gene regulation, a potential target of Ngn1 and Ascl1a in zebrafish. in Ascl1a and Ngn1 of target potential a regulation, gene deltaA family also have divergent activity in zebrafish. Moreover,zebrafish. rescue in activity divergent have also family atonal promoter led us to identify a fragment of 470pb which is which 470pb of fragment a identify to us led promoter deltaA ) after heat-shock induction, show that proneural genes of genes proneural that show induction, heat-shock after ascl1a) atonal and ngn1-/- and achaete-scute family comes from transcriptional and achaete-scute spg-/- mutants embryo, show that Ngn1 and is a target of Ngn1 and Ascl1a. and Ngn1 of target a is deltaA 426 r cnevd from conserved are atonal neurogenin1 497

Posters Posters Only zebrafish. in members family sirtuin of distribution tissue wide a illustrating profile, unique expression a showed gene Each paralogues. SIRT3 two and (SIRT1,2,4-7) sirtuins the of to one each corresponding clades, terminal well-resolved and seven identified, on throughresulted genes analysis phylogenetic sirtuin-related pattern their eight expression in resulted their analysis and The RT-PCR. relationship semi-quantitative phylogenetic their determinate members, sirtuin zebrafish the identified manner. We tissue-specific a in modulators potential testing for requirements. these expression their pattern and in genes nine perfectly sirtuin-related different all tissues of of fits identification adult time, zebrafish,first in zebrafish very orderthe tofor give andreport, awe completehere scenario Thus, needed, is organism manipulated easy model cost-effective, consolidate a and screenings molecule For activity. their modulate may that ligands discover to and sirtuins, the understand to order in investigations cellular and chemical of firestorm a in resulted that areas research many in interest much so attracted They disorders. enzymes for treatments of age-associated of set druggable potential a as appear They diseases, processes. physiological many of regulators key are such as diabetes, cancer, and neurodegenerative 498 NAD conserved evolutionary of class unique a comprise Sirtuins Brazil FaculdadeBiociências,de Pontifícia Universidade Sul.,do Grande Rio do Católica Porto Alegre, Molecular,e CelularMolecular, Biologia e de Genômica Biologia Departamento de Laboratório A. Arigony Souto,C.D.Bonan M.R. Bogo, T. Carneiro Brandão Pereira, E. Pacheco Rico, D. Broock Rosemberg, R. Dutra Dias, Expression Zebrafish as a Model Organism to Screen New Drugs Potentially Able to Modulate the Sirtuins selective andpowerfultherapiestohumandisordersassociatedwithagingprocess. new drugs based on sirtuin modulators may be tested, and in the future, could lead us into more SIRT1 isthememberwithhigherlevelofexpressioninallnineorgans. With thispanorama, 427 SIRT1, SIRT3, and SIRT5, related genes were expressed in all tissue analyzed, and analyzed, tissue all in expressed were genes related SIRT6 + -dependent deacetylases that deacetylases -dependent Gene regulation E.H. Barriga Role ofHif-1alphaintheNeural CrestCellsMigration Gene regulation nrs el, atao Chile; Santiago, Bello, Andrés 1 FONECYT 1095128 that Hif-1alphacouldbeanupstreamregulatorofsnail1bintheneuralcrestmigration. suggest data these Furthermore process. induction cells crest neural in not but migration, cells (MOATG-hif-1 heart and central nervous system development. In this study vasculogenesis, we have angiogenesis, designed two including morpholinos processes, developmental several in involved are and Hypoxia-inducible factors (HIFs) are a key regulator of the gene expression in response to hypoxia Santiago, Chile alpha of neural crest induction markers as have observed that the loss of function of Hif-1 alpha do not induce a variation in the expression was analyzed by phenotypes. morphant the rescue to mRNAmicroinjections in the 80% of the total embryos, co-injecting the morpholino MOATG-hif-1 pharyngeal in defects morphological cartilage; a observed neural have crest we cells derivate, development using embryonic the in Later of expression the aoaoi d Booí dl earlo Dpraet d Ceca Boóia, Universidad Biológicas, Ciencias de Departamento Desarrollo, del Biología de Laboratorio and mRNAs. These findings show that Hif-1alpha plays a key role in neural crest neural in role key a plays Hif-1alpha that show findings These mRNAs. snail1b 1 and A.E. Reyes alpha in situ hybridizations to neural crest cells induction and migration markers. We , a neural crest cells migration marker, is affected at 8 and 20 somites. 20 and 8 at marker,affected migration is cells crest neural a , crestin and MOSS-hif-1 and 1,2 2 ilnim nttt fr udmna ad ple Biology, Applied and Fundamental for Institute Millennium foxD3 and Ap2 ) to generate the loss of function of function of loss the generate to alpha) Alcian blue stain. The defects observed were rescued, at the 1-somite to 3-somites stages, however, The loss of function of Hif-1 alpha Hif-1 of function of loss The 428 alpha with both hif-1 and alpha hif-1 499

Posters Posters n i te al ad dl lf wl hl t udrtn mcaim o mmcig endocrine mimicking of mechanisms understand to help will chemicals. life adult and early the in and as and development of during understanding Current fish. agonist adult in antagonist 17β-estradiol or agonist a as acts 4-nonylphenol ligand estrogenic synthetic the iii) context; cellular specific the to restricted is 17-βestradiol of to response expression in genes altered ii) zebrafish; adult of of organs absence in in and embryos receptors in estrogen expressed i) are ligand that are conclusions The testis. and brain intestine, heart, to ligand in the organs tested (except in heart) while the expression of the fish, adult In accumulated. ERs active two expressed during the embryonic stages and in presence of 17β-estradiol the mRNA levels of the 500 genes, receptor three the of Two fish. adult of organs in expressed were receptors estrogen three all that show fish individual from Data PCR. time real using quantified were levels expression the ERs zebrafish three the of each for primers specific testis, ovary, brain, heart, eye, intestine, Using (liver,isolated. were fish adult exposed ligand or exposed not of kidney) and bladder swim gill, organs several of and fertilization) post hours and (ERbeta2) esr2a denoted of are they and receptors estrogen of isoforms three are there zebrafish numberIn a in 17β-estradiol. to response in involved factors transcription as function mammals in identified been proteinshave regulatory of group aphysiological processes. The estrogen receptors (ERs), of which one alpha and one beta isoform encompass receptors nuclear The ofBiosciences andNutrition,Department Karolinska Institutet, Stockholm, Sweden IN ZEBRAFISH ADULTM. Andersson Lendahl,G.Chandrasekar, A. Archer, andJ-Å.Gustafsson AND EMBRYONIC IN ABSENCE AND PRESENCEOFLIGAND EXPRESSED RECEPTORS 17β-ESTRADIOL 429 (ERbeta1). Total RNA of embryos (<3, 6, 12, 24, 48, 72, 96 and 120 and 96 72, 48, 24, 12, 6, Total(<3, (ERbeta1). embryos esr2b RNAof expression was not regulated in response in regulated not was expression esr2b and esr1 gene function in development in function gene esr bt not but esr2b, esr2a was altered in liver, Gene regulation (ERalpha), esr1 were esr2a esr Life Sciences, Sördertörn University College,University Sciences,Huddinge, Sördertörn Sweden;Life of generallipiddistribution,reducedyolkutilizationandcartilagedefects. impairment as such embryos zebrafish in defects developmental induces oligonucleotids sense anti- morpholino by expression Lxr of down knock the Indeed, role. developmental a suggests pattern expression temporal and ubiquitous the Lxr of role metabolic a to addition In dpf. 3 at expression of ubiquitous a observe zebrafish. We in Lxr of role metabolic conserved a suggesting pathways, synthetic LXR ligand, GW3965, induces the expression of genes involved all in lipid fish and adult cholesterol In LXR-ligands. examined classical organs to express response in activity transcriptional has it that shown have we and alpha LXR mammalian with similarity sequence high shares protein Lxr zebrafish we choose to explore LXR function during embryonic development in the zebrafish model. The adulthood. at reasons, these For observed suggest. to reasonable is age, adult until manifested generally not possibly period, this are mice during in LXR of role a LXRs development, during cholesterol of importance of the However,considering testicular deletion of maintenance targeted for of and Effects degeneration, neuron function. motor of prevention for beta importance the LXR shown of have studies recent Moreover, metabolism. glucose and lipid regulate cholesterol metabolism and are potential drug targets for treatment of atherosclerosis. They also The Liver-X-receptors (LXRs) alpha and beta are activated by oxysterols. LXRs are implicated in and Cell Signalling, University ofHouston, Texas, US 1 A. Archer Effect ofLiver-X-Receptor (lxr)knock down duringdevelopment inzebrafish Gene regulation Dept of Biosciences and Nutrition,and NOVUM,Biosciences of Institutet,,Sweden;KarolinskaDept Huddinge 1 , S.Kitambi lxr mRNA during zebrafish development that is particularly pronounced in the liver 1,2 , J.-Å.Gustafsson lxr. Treatment of zebrafish at adult stage or during development with the 1,3 and A. Mode 1

3 Center for Nuclear Receptors Nuclear for Center 430 2 School 501

Posters Posters the demonstrate for first time that paralog group 4 genes play a significant role in haematopoiesis and angiogenesis. in role significant a play genes 4 group paralog that time first for demonstrate blood pooling was observed in the cardiac region, eye and in yolk. Taken together, these results as such markers arterial the of expression of Remarkably, circulation. blood characterized by the loss of of arterial endothelial cell lack identity in dorsal aorta, and as shown by the sprouting, lack vessel and intersomitic development in (ISV) failure bifurcation, aortic anterior of interruption and atrophic the aorta including dorsal angiogenesis, trunk and where development vascular embryos, in alterations profound Fli:EGFP in results Tg in examined promoter was Vasculature development. during vessels blood of formation the with connected tightly is cells endothelial of differentiation The malformed were or abolished. morphants the in cartilages pharyngeal the that showed staining blue Alcian islet-1 expressing progenitors that gives rise to somatic motoneurons and branchio motoneurons. ee hv be ipiae i te differentiation the in implicated been have genes 502 induction the by vertebrate characterized the is in Haematopoiesis elucidated. be to remains rhombomeres the within lineages specialized However, the interaction of Hox genes with transcriptional program to specify distinct neuronal with neurons of of Hox classes combinations genes has and revealed individual their many of role in loss specifying functional rhombomere identity the contains and hindbrain, neuronal developing development. the system In properties. nervous central vertebrate The School ofBiologicalSciences,Nanyang Technological University, Singapore A. Anusha Amali, T. S.Fiona,C.Winkler andM.Featherstone of Hoxgenes Abnormal neurogenesis, angiogenesis and haematopoiesis in zebrafish lacking group 4 paralog n cmie kokon f 4 of knockdown combined and Wesingle that haematopoiesis. demonstrate and angiogenesis neurogenesis, in defect early an Here, through the morpholino knock down of group 4 paralog of Hox genes, we have identified and spatialexpressionoftranscriptionfactors,suchastheHoxgenefamily. temporal by regulated largelycoordinately is are development embryonic cells of much endothelial general, In and unknown. haematopoietic to haemangioblast the of differentiation the a common embryonic precursor, termed the haemangioblast. The genetic cascades that regulate vascular progenitor cells. The first haematopoietic and endothelial progenitors are derived from

431 eventual differentiation of these progenitors to form the peripheral the form to progenitors these of differentiation eventual drives expression of enhanced green fluorescent. Loss of hoxc4a and d4a and hoxc4a of Loss fluorescent. green enhanced of expression drives th

ru o Hx ee rsls n ls o oi2 nx . and 2.2 nkx olig2, of loss a in results genes Hox of group of ventral mesoderm to form haematopoietic stem cells and cells stem haematopoietic form to mesoderm ventral of and flk-4, flk1

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blood lineages. Several lineages. blood Gene regulation opat are morphants . Also flt4. function fli1 into zebrafishembryos. Atthemeeting,wewillgiveaprogressreport. an introducing by the around sequence genomic of 176kb contains which BAC a modified have we a basal promoter of the using comparisons genomic interspecies sequence alignment programme MLAGAN. These elements were individually in cloned in front of elements noncoding we conserved conserved evolutionally for be searched may elements such that Assuming region. proximal promoter the outside elements identified regulatory for searched were have Secondly, we fish. founders wild-type Transgenicwith crosses in up. grown Embryos were expression integration. GFP genome transient facilitate displayed to that used were sequences transposon the and embryos a in gene reporter the in of upstream and sequences tested brain have the we zebrafish Firstly, approaches. arches, different branchial three used the have we placode, olfactory controlling the elements regulatory Tofind intestine, tissues. haematopoietic the retina, the in of regulation about known the transcriptional is regulation Little of placode. a olfactory the in and expressed retina neural be the to including tissues, known other also of is number It intestine. the and brain the blood, the in differentiation and proliferation cell stem/progenitor in role essential an plays c-Myb factor transcription The Institute ofGenetics, SchoolofBiology, University ofNottingham, Queen’s MedicalCentre, UK M. GeringandJ-C.Hsu Transcriptional RegulationoftheStemCell/ProgenitorGenec-myb Gene regulation c-myb locus. Upstream sequences were amplified by PCR and cloned in front of a in zebrafish embryos. In zebrafish embryos, zebrafish In embryos. zebrafish in c-myb reporter gene into the into gene reporter rfp transposon vector. The resulting constructs were injected into zebrafish into injected were constructs Tol2vector.resulting transposon The c-myb in the Tol2 reporter vector and tested in transgenic zebrafish. Finally, c-myb. This is why we are investigating the transcriptional locus. The modified BAC has been injected been has BAC modified The locus. c-myb expression in zebrafish in expression c-myb expression is detected is expression c-myb 432 locus c-myb 503 gfp

Posters Posters for degradation of PTC-containing mRNAs in zebrafish embryos using the using embryos zebrafish in mRNAs PTC-containing of degradation for zebrafish the vertebrate, basal a in investigated pathway we this NMD of relevance physiological and evolution the on insight further Togain the courseofevolution. the NMD pathway or to the inhibition of an unknown function that Upf1 or Upf2 acquired over in seen phenotypes the whether of question the organismraises vertebrate a of context the in effectors NMD additional these of Smg1, Smg5–7 have not been described in vertebrates. The lack of information embryonic development on in the the mouse. Remarkably,importance phenotypes associated with the depletion of in Across species, the importance of NMD effectors varies. Indeed, NMD effectors are not essential ubiquitously expressedthroughoutearlyzebrafishembryogenesis. 504 N. Wittkopp and survival Nonsense-mediated mRNA decay effectors are essential for zebrafish embryonic development splicing andNMDhasbeenmaintainedinvertebrates,butlost infungiandinvertebrates. of coupling the that suggest and embryogenesis vertebrate for essential are effectors NMD that and invertebrates to contrast in but fungi, NMD is coupled to splicing in zebrafish.plants, Our results, together with previous studies show and mammals in as Thus, boundaries. exon-exon downstream by stimulated strongly is NMD trigger to PTC a of ability the that patterning demonstrate we embryos, early Smg6-depleted or development, Smg5- Upf2-, embryonic pathway. NMD cells active cultured an Using requires embryogenesis zebrafish in that suggesting on observed are impact phenotypes severe Similar viability. a and has depletion its Moreover, 1 mutants in the the thaliana, orthologs encodes genome zebrafish the upf1, that of show we here NMD; for essential as identified In progression. cycle cell transduction and thereby signal development, transcripts, including processes wild-type biological of of range 1–10% broad a regulates influencing NMD function, protective this to addition In proteins. truncated of accumulation the from resulting mRNAs effects dominant-negative potentially anddegrading from cells eukaryotic protects bydetecting pathway NMD the so, doing expression In mutations. gene nonsense containing of fidelity the quality-control ensures mRNA conserved that a mechanism is (NMD) pathway decay mRNA nonsense-mediated The II, MPI, Tuebingen, Germany or cerevisiae S. 433

upf2, 1 and UPF1 , E.Huntzinger UPF3 gene result in strong phenotypes. smg1, . In contrast, In . elegans C. and two and smg5–7 genes are essential for embryonic viability, and loss-of-function viability,and embryonic for essential are genes SMG7 1 , C.Weiler 2 III, MPI, Tuebingen, Germany metazoa, and upf1 paralogs, which are provided maternally and are and maternally provided are which paralogs, upf3 1 , M.Sonawane ee gns (upf1–3, genes seven knockouts can be ascribed to the inhibition of inhibition the to ascribed be can knockouts upf2 is an essential gene in gene essential an is upf1 Danio rerio rerio Danio Upf1 and 2 andE.Izaurralde and show, that Upf1 is required is Upf1 that show, and and smg1 upf2 are also essential for early A. In . melanogaster D. Gene regulation 1 hv been have smg5–7) golden b1 mutant. Similar event was saw in mice subjects a CR. We can conclude that our work is the first that first the is work our consolidate thezebrafishasmodelofSIRT1that expressionbytrans-resveratrol. conclude can We CR. a subjects mice in saw was event Similar minutes. 60 in level, control the to returning minutes 30 and 15 in decrease a saw we specific, tissue- is that However,expression, SIRT1literature. gene the with agreement in are results our PPAR-and targets PGC1- α the expression Regarding gene significant. not was increase the but γ 60 minutes (1,01 SD ± 0,03 p<0,05). The similar results are observed in PPAR-γ gene expression, the control in 15 and 30 min treatments, but was observed an significantcontrol increase(p<0,01 and ofp<0,05). expressionThe expression in means of PGC1-α was not significant different than The decrease of gene expression of SIRT1 in 15 and 30 minutes are statistically significant versus respectively.minutes, 60 and 30 15, control, in 0,09) ± (SD 0,67 and 0,07) ± (SD 0,56 0,02), ± significant. The means of expression of SIRT1 in liver of zebrafish was 0,75 (SD ± 0,08), 0,36as TukeyP≤0.05 by ANOVA,one-way (SD considering followed by test, compared statistically and was determined by optical densitometry using ImageJ1.37. Results were expressed as mean ± S.D on 1.0% agarose gel with GelRed®. The relative mRNA abundance of each gene versus designed, and PCR optimal conditions were determined. Reactions were performed and analyzed synthesized through semi-quantitative RT-PCR. SIRT1, PGC1-α, PPAR-γ and Total cDNAmin. were and 60 species (n=5) or liver 30 zebrafish adults from RNAisolated was 100 in dissolved (5mg/L solution resveratrol containing L) (2 aquariums PPARγ by resveratrol in the of this work was to demonstrate the and gene expression simplicity of the between SIRT1 balance and its a two targetsshows complexity,PGC-1 SIRT1about work one is there actually but manipulated, easy aim The modulation. it because organism model used an is ) rerio (Danio mechanism.. this by CR mimic partially to proposed been SIRT1has modulate and to Bonan, MaurícioReisBogo, A. Arigony Souto Sul - Brasil; * . Schirmer H. Regulation ofSIRT1, PGC-1αandPPARγ by Resveratrol inLiverZebrafish of Gene regulation such as the transcriptions factors PGC-1 nuclear deacetilase, important an protein, well-studied most the become has 1 SIRT life. of domains all in of modulation activity,the deacetilase NAD+-dependent with enzymes (SIRT),of sirtuin family conserved found a is proposed mechanisms the of One organisms. model of variety wide a in life-span extends and diseases age-associated of appearence the slows (CR) restriction Calorie Laboratório deBiomedicina, Prédio Branco(Brasil) Laboratório de Química e Produtos Naturais - Faculdade de Biociências - PUCRS, Rio Grande do # Curso de Biomedicina - Instituto de Ciência da Saúde - Centro Universitário Feevale, whose substrates include proteins primarily involved in transcriptional regulation, transcriptional in involved primarily proteins include substrates whose *# T Crer Bado eer, . ahc Rc, . rok oebr, . D. C. Rosemberg, Broock D. Rico, Pacheco E. Pereira, Brandão Carneiro T. , zebrafish liver. α and PPARγ. The molecule of For the treatments, animals were introduced into test 434 trans-resveratrol is known L ethanol), during 15, during ethanol), µL β-actin primers were Zebrafish β-actin α and 505

Posters Posters 506 Muscle during Genes Chain Heavy Myosin Development of Medaka Sarcomeric of Expression Spatio-temporal The expressed in zebrafish embryos in a similar manner as found in medaka, suggesting common suggesting regulatory functionsofthe5’-flankingregionfishMYHs inexpressionduringdevelopment. medaka, in found as manner similar a in embryos zebrafish in expressed endogenous of those recapitulated fish transgenic in RFP and GFP of patterns expression specific spatio-temporally The eggs. fertilized into them genes encoding green of fluorescent protein region (GFP) and5’-flanking redthe fluorescent proteinwhich (RFP) andin introduced constructs each several of prepared region 5’-flanking the by regulation transcriptional the analyzed we Then, (dpf), dayspost-fertilization 2 from whereas the other two in whole myoseptum myotome from 3 dpf and intensely expressed at hatching stage. horizontal in specifically expressed was one type muscle skeletal fast by RT-PCR and First, we randomly cloned muscle development in medaka Here, we analyzed the expression of sarcomericdifferent contain muscles vertebrate that MYH genes( s) whichshowspatio-temporallydifferentexpressionpatterns. known well is It by actin. protein to motor bound a when ATPas splitting functioning chain heavy former the with chains, light four and (MYHs) Class II myosin is a major contractile protein in muscle fibers and composed of two heavy chains Graduate Schoolof Agricultural andLifeSciences, The University of Tokyo Y. Ono,S.Kinoshita,Watabe by Transgenic Constructs 435 in situ hybridization at several developmental stages. As a result, three embryonic Oryzias Oryzias latipes MYHs from embryos and larvae and analyzed their expression patterns wr peoiaty xrse drn msl development: muscle during expressed predominantly were MYHs Oryzias latipes Oryzias to investigate their relation to muscle formation. Are Regulated by Their 5’-flanking Region as Revealed MYHs and their transcriptional regulation during s. Furthermore, these constructs were constructs these Furthermore, MYHs. was fused to the marker the to fused was MYH Gene regulation . We MYH. S. Moleri The hmgbgenefamilyinzebrafish: sixmemberswith overlapping expressionpatterns Gene regulation oooos o mammalian to homologous Through expressed duringembryogenesisbuthaveaveryrestrictedexpressionpatterninadults. expression patterns: in fact, HMGB1 is almost ubiquitous their in while and HMGB3, HMGB2 in shorter and and HMGB3 HMGB1 in are longer is largely which tail, acidic the of length the in lie members family HMGB of differences main The tail. C-terminal acidic an by and B) and A box (HMG domains binding DNA tandem homologous two by characterized all HMGB3, and HMGB2 HMGB1, members: conserved highly three comprises family as HMGB the mammals roles In key also have and repair secreted proteins. and replication transcription, including numerous processes regulate HMGBs nuclear chromatin. the to plasticity functional and structural imparting it, bend sharply and specificity sequence limited very with DNA the of groove that minor factors the contact binding chromatin abundant very are proteins Box) Group Mobility (High HMGB degli StudidiMilano, Italy 1 the mammalian homologs. zfhmgb homologs. mammalian the canonical the display proteins zebrafish structure of HMGB factors, i.e. the sixtwo HMG boxes and the acidic tail, The and are highly related to analysis. syntheny and trees phylogenetic of gene and named them hmgb1a/1b, domains although their expression levels are differentially modulated. Later in the development expression overlapping largely display genes zfhmgb all dpf, 1 Until levels . expression in differ all that show we RT-PCR Real-Time and We analyzed the expression of the translated. not is one first the which of exons five contain and introns four by interrupted are they fact, in the central nervous system while system nervous central the that the co-orthologous genes are mainly expressed in the same cell types and therefore are therefore and types cell same the probably implicatedinthesamebiologicalfunctions. in expressed mainly are genes co-orthologous the that Dipartimento di Scienze Biomolecolari e Biotecnologie, e Biomolecolari Scienze di Dipartimento 1 , G.Cappellano bioinformatic analyses, we identified six zebrafish cDNA/genomic sequences cDNA/genomic zebrafish six identified we analyses, bioinformatic hmgb2a/2b hmgb1a, and 1 , D.S.Horner HMGB hmgb3a hmgb1b, hmgb2a, hmgb2b, hmgb3a and hmgb family genes in zebrafish embryos. By hmgb3b genes also possess the typical the possess also genes ee. e on to ootoos o ec mammalian each for co-orthologs two found We genes. are restricted to specific regions of the embryos, mainly in 1 , G.Gaudenzi is barely detectable. Our data support the hypothesis the support data Our detectable. barely is rncit sae mtra oii, u they but origin, maternal a share transcripts hmgb 2 , F. Cotelli 2 Dipartimento di Biologia, Università Biologia, di Dipartimento intron-exon structure: intron-exon HMGB 2 436 andM.Beltrame hmgb3b, on the basis in situ hybridization 1 507

Posters Posters Financial Support: CNPQ; FAPERGS; PUCRS. organism. model the this in genes target sirtuins elucidate and sirtuins both over to consumption alcohol chronic of help effect could organs other pathern, expression tissue-specific a has gene SIRT1 Since zebrafish. in metabolism alcohol to due changes metabolic by mediated be could levels decreased expression at 28 days treatment. Our results suggest that the increase in SIRT1 mRNA a with pattern, similar a demonstrated 0.05) ± 0.4855 0.05*; ± 0.7663 0.08*; ± 0.7670 0.05; ± (0.6196 PPAR-γ and 0.04) ± 0.4244 0.02; ± 0.8763 0.01*; ± genes, 0.8130 target SIRT10.03; ± of (0.8736 expression PGC-1α The respectively). exposure, of days 28 and 0.03 14 ± 7, 0.8885 control, 0.01*; - ± 0.9917 0.07*; ± 0.9849 0.07; ± (0.7842 treatments all in gene SIRT1 Tukey of activation showed patterns expression Gene (*). significant as P≤0.05 considering test, by ANOVA,one-way followed by compared statistically were (mean±S.D) Results ImageJ1.37. versus gene each of abundance mRNA determined. Performed reactions were analyzed on 1% agarose gel with GelRed with gel agarose 1% on analyzed were reactions Performed determined. PPAR-and PGC-1α, γ SIRT1, RT-PCR. semi-quantitative through synthesized species cDNA and (n=5) liver zebrafish adults nitrogen. liquid in frozen livers from RNA their isolated Total and was euthanized were fish the exposure, after Immediately days. 28 and 14 7, during 0.5% ethanol with treated were Animals and 28 days in the gene expression of SIRT1, PGC-1α and PPAR-γ related-genes as the zebrafish. Thisin work aimed tozebrafish study the role of liver. chronic alcohol consumption during 7, 14 these targets, specially using sirtuin cost-effective, easy manipulated with and consolidate Sincestudied model organisms,be could consumption. strategies therapeutic new alcohol enzymes, of set chronic druggble a as by appears family affected were levels protein and mRNA both since levels translational and transcription the at regulated were level SIRT1protein in changes alcohol-induced the that indicates evidence accumulating Furthermore, fibrosis. and necrosis data indicate that chronic ethanol feeding develops to alcohol liver diseases, including steatosis, regulating proliferator-activated or (peroxisome histones PGC-1α receptor- modifying as regulators, direct is transcriptions either Recently, several of by metabolism. activity regulator the and master aging metabolic in a roles as emerging physiological important play that deacetylases 508 NAD conserved evolutionary of class unique a to belongs (SIRT1) 1 Sirtuin RS -Brazil Laboratório de Biologia Genômica e Molecular, Faculdade de Biociências, PUCRS, Porto Alegre, A. Arigony Souto,M.R.Bogo. T. Carneiro Brandão Pereira, Long-term alcoholtreatmentincreasesSIRT1 geneexpressioninzebrafish liver 437 coactivator1 γ ). Previous receptor-proliferator-activatedγ). (peroxisome PPAR-γ and α) -actin primers were designed, and PCR optimal conditions were conditions optimal PCR and designed, were primers β-actin H. Schimer, E. Pacheco Rico, D. Broock Rosemberg, C. D. Bonan, -actin was determined by optical densitometry using densitometry optical by determined was β-actin Gene regulation ® . The relative The . + -dependent lo eut i ls o te eaornhas We ijce into injected When ceratobranchials. the of loss in results also hand2 constructs into wild type embryos leads to mispatterning of the ceratohyal, while our of all of injection that found far thus have we ongoing, is analysis our While development. cartilage analyze to blue alcian with stained and fertilization post days 5 at fixed were Embryos and type wild into injected was constructs these To constructs. (ΔR100D)- and HLH (ΔF110P)- EnR)-hand2 hand2 mutant constructs. Capped mRNA generated from each of - basic created we dimerization, HLH (Engrailed or binding DNA requires action Hand2 super-repressorwhether examine and (VP16)- super-activator created of analysis our For binding. DNA or dimerization bHLH whether Hand2 acts as a transcriptional activator or repressor and whether this function requires determine to begun have we activity.zebrafish, in pathways these understand better to order In its about known is pathway,little signaling Ednra/Dlx the of mediator primary a be to appears Hand2, a basic-Helix-Loop-Helix (bHLH) transcription factor expressed in the mandibular arch, While development. jaw lower in defects of severe and archloss mandibular the within in identity NCC results pathway Ednra-Dlx5/6-Hand2 the of Loss (Ednra). receptor endothelin-A the the form into a lower eventually jaw relies on multiple signaling cascades, including and one mediated by signaling from arches arch pharyngeal first the of portion mandibular the of Development abnormalities. pharyngeal craniofacial to leading the development NCC in defects jaw,with populate the of tissue connective (NCC) and cartilage bone, cells crest neural Cranial Aurora, USA Medicine, Biology,Dental Craniofacial of Denverof ColoradoSchool of Department University J. IkléandD.E.Clouthier Signaling mechanisms regulatinglower jaw development Gene regulation binding. and posterior pharyngeal skeleton and that some of these functions occur independently of DNA anterior the of development the in differently functions Hand2 that appears it data, preliminary of injection ventral cartilages. Injection of Δ R100D EnR-hand2 ece al eta criae, while cartilages, ventral all rescues rescues both the anterio- and posterio-ventral cartilages. Based on this on Based cartilages. posterio-ventral and anterio- the both rescues VP16-hand2 also only rescues the anterio-ventral cartilages, while han s6 hand2 zebrafish embryos at the 1-2 cell stage. cell 1-2 the at embryos zebrafish as an activator/repressor, we activator/repressor, an as hand2 Δ F110P han 438 ny ece te anterio- the rescues only s6 mutants, we find that find we mutants, hand2 Δ R100D 509

Posters Posters 510 could beduetoanoverexpressionofnucleolarproteinsinvolvedinthep53pathway. mouse cells respectively.and These results suggest human that an activated p53 response in L11-deficientin embryos p53 activates Bop1 and gnl3), of homolog human (the Nucleostemin of coincided with the activation of p53 response in the morphants. Interestingly, aberrant course expression analysis at various stages post fertilization indicated that overexpression of displayed morphological abnormalities and an increased p53 localization. Furthermore, a time- of enrichment showed hybridization embryos. wild-type in that to similar was genes protein nucleolar other of expression the RT-PCR revealed a selective upregulation of for identified already bop1), and their role pes, in ribosomal assembly,gnl3, were ncl, analyzed in this bxdc1, study. rrs1, Quantitative and semi-quantitative bap28, wdr55, (wdr36, proteins nucleolar nine L11-deficient of in (morphants). total genes embryos A protein nucleolar several of expression the examined we response, p53 the RP activates an deficiency how investigate to RPL11 of knockdown that demonstrate activates the data p53 pathway previous in zebrafish, our which However, is contrary to loop. the cell regulatory line-based results.MDM2 In an effort p53- the of inhibitor major a RPL11 as identified have lines cell in Studies stress. ribosomal or mutations in nucleolar proteins activate the p53 pathway presumably through a nucleolar stress or RP an of decrease the by induced biogenesis ribosome of Recent impairment that nucleolus. indicate findings the within assembly pre-ribosomal and processing pre-RNA in role critical play proteins nucleolar Several cytoplasm. the in ribosome mature of formation the with ends and nucleolus the in begins that process multi-step controlled tightly a is biogenesis Ribosome Frontier ScienceResearch Center, University ofMiyazaki, Japan A. Chakraborty, T. Uechi,H.Torihara andN.Kenmochi modulatory nucleolarproteins Ribosomal protein L11 (RPL11) deficiency in zebrafish leads to a selective upregulation of P53- 439 and gnl3 gnl3 and in the head region of the morphants that morphants the of region head the in bop1 bop1 transcripts in the morphants whereas Gene regulation gnl3 and In situ In bop1 targeted to morpholino (MO) injections or mutant embryo analysis. Injections of translation-blocking MOs oligonucleotide antisense through analysis knockdown by either out carried was analysis (LOF) function of Loss stages. similar while at embryo (hpf), the throughout fertilization ubiquitously levels post low at hours expressed 48 and 24 both at domains arch branchial and development. craniofacial in involvement and Gene expression analysis of candidate multiple that unknown. currently hypothesize are We development craniofacial zebrafish in family gene this of members other of roles potential The cartilages. ceratobranchial posterior missing including of the branchiostegal rays. We are currently examining markers for neural crest specification, crest patterning andmigration,inadditiontobonedifferentiation. neural for markers examining currently are We rays. branchiostegal the of 1 L. Kwok prdm genesinzebrafish craniofacial development Gene regulation PR/SET domain to mediate protein interactions, and a variable number of DNA-binding zinc DNA-binding domains. of number finger variable a and interactions, protein mediate to domain PR/SET The zebrafishprdmgenefamilyiscomprisedofaset17genesthatallcontainanN-terminal Biology, University ofColorado Denver - MedicalCampus,Anschutz Aurora, USA prdm5 plate. ethmoid and cranii trabeculae the of mispatterning through neurocranium shortened a in of fingers zinc binding DNA the of translation inhibit to MO splice-blocking of el ilg, tm el ad eeomn Gaut Program, Graduate Development and Cells Stem Biology, Cell . These genes are expressed in temporal-spatial patterns consistent with putative with consistent patterns temporal-spatial in expressed are genes These prdm16. hi61 1,2 mutant embryos also exhibit neurocranial shortening, in addition to reduction or loss or reduction to addition in shortening, neurocranial exhibit also embryos mutant , D.E.Clouthier prdm3 and genes are involved in the patterning and development of the zebrafish face.zebrafish the of development and patterning the in involved are genes prdm a ebr f the of member a prdm1a, prdm16 did not result in craniofacial cartilage defects. However, injections 2 andK.B. Artinger prdm genes identified three novel targets – and prdm3 2 ee aiy ehbt caifca defects craniofacial exhibits family, gene prdm

are expressed in specific neural specific in expressed are prdm16 2 440 eatet f Craniofacial of Department prdm3, result prdm16 is prdm5 prdm5 511

Posters Posters 512 autoregulation andexpressionlevelsindifferentspatiotemporalsettings. of requirement the on informative prove will approach an such that Weanticipate locus. functional modules of this element and zinc finger nuclease technology to alter the endogenous of part recapitulates zebrafish the transgenic in the and that enhancer, transcriptional demonstrated a have as We acts sites. sequence binding PTF1 putative ofzebrafish three contains that upstream anamniotes, sequence a non–coding identified have We The domains. progenitor enhancer’s pancreatic activity requires multipotent two to PTF1 binding significantly sites, and which are tissue, thought acinar to mediate and autoregulation. neural to expression transgene directs it expression; Ptf1a endogenous the recapitulates partially Ptf1a in these domains is less well understood. A conserved non–coding sequence upstream of mouse endocrine tissue of the pancreas and hindbrain glutamatergic neurons, although its requirement of precursors in expressed also is complex PTF1 the of component PTF1a The ofretina. and cord spinal fate dorsal cerebellum, cell the of of neurons inhibitory determinant GABAergic the critical and pancreas a exocrine the is PTF1 complex factor transcription heterotrimeric The Cell andDevelopmental Biology, University ofPennsylvania, USA E. Pashos andS.Fisher Mutational analysisofptf1afunctionandtranscriptional regulation 441 ptf1a noeos xrsin W ae sn tasei ehne aayi t dset the dissect to analysis enhancer transgenic using are We expression. endogenous cnevd among conserved ptf1a, Gene regulation ptf1a display distinct and complex patterns of tissue-specific expression, and are found in of kidney, levels Expression are found retina. and and brain, expression, lung, tissue-specific of patterns complex and distinct display derived by transcription from two different promoters and alternative splicing. The three isoforms encephalocele. occipital and degeneration macular and vitreoretinal by characterized disease recessive autosomal an Syndrome, Knobloch to lead the in mutations membranes; basement most of component a is collagen XVIII Typefunction. enhancer of predictor imperfect an proven has conservation sequence although non–coding for sequence. It particularly is commonly sequence, hypothesized DNAthat evolutionary primary conservation predicts from functional DNA, function predict and understand to The completion of multiple vertebrate genome sequences has presented an important challenge Biology, University ofSao Paulo, SaoPaulo, Brazil computational alignmentalonefallsshort. where cases of number large the in elements cis–regulatory conserved reveal can conservation More generally, we show that combining functional data with targeted analyses for phylogenetic and point to additional sequences that may contribute to complex diseases involving of regulation and function biological sequence. the into enhancer insight important human provide results orthologous Our the with overlap functional demonstrated we these, of one for enhancers; zebrafish corresponding the predict hypothesized we which sequences, teleost and mammalian between alignments revealed sequences enhancer positive on analysis sequences did functioned appropriately in used zebrafish transgenics. we Additional post–hoc computational algorithms the the at teleosts to Although human from conservation non–coding analysis. detect not sequence comparative for approach on single relying of a risk the demonstrates it Importantly, sequence. longer the in element regulatory algorithms. In one pair, only the shorter sequence was an active enhancer, suggesting a negative largely different sequences; only three pairs of CNSs, partially overlapping, were found with both pronephros, blood vessels, gut, cartilage and liver. Significantly, VISTA and PhastCons identified transcription consistent with control endogenous that elements four identified We zebrafish. transgenic in transcription specific regulate regulating (CNSs) sequences elements non-coding selected were conserved twenty enhancer algorithms, PhastCons and VISTA potential the Using evaluate functionally to of Therefore, 2. type diabetes identification ofand thehepatocarcinoma regulatory regionsto willpredisposition provide insightin into normaland andvasculogenesis, pathogenic regulation in 1 Institute of Genetic Medicine, Johns Hopkins University, Baltimore, USA; Functionally conserved cis-regulatory elements of COL18A1 identified through zebrafish through E. Kague identified COL18A1 of transgenesis elements cis-regulatory conserved Functionally Gene regulation el n Dvlpetl ilg, nvriy f enyvna Piaepi, USA; Philadelphia, Pennsylvania, of University Biology, Developmental and Cell expression. We have employed an efficient system of transgenesis in the zebrafish the in transgenesis of system efficient WeCOL18A1an expression. employed have 1 , S.Bessling from the interval encompassing interval the from silico in 2 , J.Lee 2 , M.R.Passos-Bueno col18a1, in tissues including retina, pronephric duct and COL18A1 3 , andS.Fisher has 43 exons; three isoforms are isoforms three exons; 43 COL18A1has and tested for their ability to ability their for tested and COL18A1 are thought to be clinically important clinically be to thought are 1 442 locus, the human the locus, COL18A1 transcription. COL18A1 3 Genetics and Evolutive gene COL18A1 COL18A1. COL18A1, 2 Nathans 513

Posters Posters 514 functions areachievedthroughsomeofthetargets. a major transcriptional modulator. Furthermore, we are starting to investigate how specific pax6 encode proteins with DNA-binding or transcription factor activity, reinforcing the idea of Pax6 genes andhave as downs, knock target pax6 putative induced vertebrate morpholino as such methodologies using experimentally,zebrafish, some one hundred in validated than more identified have We the target genesunderdirectregulation. and (ECRs) sequences conserved evolutionarily predict to models probabilistic and sequences genome (TFBS), site binding factor transcription of knowledge bibliographic as such tools, and by regulated transcriptionally data combined of use makes procedure This (TFs). are DNAfactors given transcription any binding that genes identify to procedure bioinformatic a developed Using comparative genomics, we are investigating transcriptional regulation by pax6. We have vertebrates. of regulation Pax6 Furthermore, quantitative evolution. across and spatiotemporal complex expression at by This the transcriptional pancreas. level. and achieved Pax6 transcripts system encode proteins nervous that is are central highly conserved eye, multi-tasking the tissues: developing different in fates cell The General Hospital, Edinburgh, UK Unit,WesternGenetics Human Anaysis,Systems MRC Biomedical & Section Genetics Medical P. Coutinho,D.A.Kleinjan,V. vanHeyningen Deciphering thepax6 Transcription Network 443 pax6 gene has been defined as a selector gene, responsible for the determination of multiple hybridization and ChIP. This set of loci is highly enriched for genes that genes for enriched highly is loci of set ChIP.This and hybridization situ in i-euaoy eune ae lo el osre across conserved well also are sequences cis-regulatory Gene regulation pax6 be verifiedbychromatinimmunoprecipitation(ChIP)andgeneticknockdowninzebrafish. electrophoretic next will hypothesis from This G. and F sites to binds Kreisler that results show (EMSA) assays shift mobility Preliminary (TGCTGANT/CCNGNN). sequence recognition maf Sequence alignment has shown that conserved sites of regulator negative a is zebrafish) in F valentino and mouse and the G in (Kreisler factor show transcription family maf zebrafish some the of member homology a the that suggests to evidence Genetic of the analysis consensus similar A visible. clearly hoxd4a neuralenhancerwillbecarriedoutinzebrafishembryos. are rhombomeres when (hpf) fertilization at embryos zebrafish in assessed be next no will mutants These had expression. together,gene and individually G, C, delayed RARE B, except A-G sites in mutation compound A, but strength, enhancer on effect sites conserved the in mutations that showed Site-directed mutagenesis of the mouse enhancer and functional and tests in differentiating P19 cells G and C B, A, sites remaining identify thetranscriptionfactorsactingthroughneuralenhancer.the characterize will I studies, my of phase this complete Tocharacterized. previously been have RARE the and D-F Sites (RARE). element response acid hoxd4a neural enhancers revealed eight blocks of homology, denoted as sites A-G and a retinoic and 7 in the developing hindbrain. Sequence comparison between mouse and zebrafish of border anterior highly of the vertebrate are They how understand morphogenesis, to development. aims lab Featherstone organogenesis, The gland limbs, mammary and and conserved throughouttheanimalkingdomandtheirexpressionistightlyregulated. production tubercle shaft genital hair axis, hematopoiesis, anterior-posterior the of Hox genes encode homeodomain-containing transcription factors, required for correct patterning Biological Sciences, Nanyang Technological University, Singapore S. Y. LerandM.S.Featherstone Characterization oftheHoxd4 neural enhancer Gene regulation Hoxd4 neural enhancer. The xrsin ewe tasety iie sget (hmoee) 6 (rhombomeres) segments divided transiently between expression Hoxd4 Hoxd4 neural enhancer is required for setting the expression is controlled through the study the through controlled is expression Hox Hoxd4hindbrain. embryonic the in 444 16 hours post- hours ≥16 Hoxd4/ Hoxd4 515

Posters Posters by cold temperature and seemed to be correlated with the number of repeated sequence in CR. temperature The expression of the cytochrome oxidase subunit I seasonal gene in some local population was induced the and environment cold the number variation. In cold (5°C) to acclimated fish, mRNA TR levels varied among related mitochondria coding genes. the data that certain indicated the habitat to corresponding correlated in data meteorological and number TR the between comparisons Comprehensive repeats. no had regions, tropical inhabiting species, other two the while varied, highly was region control mtDNA in (TR) units nucleotide 516 in frequent tRNA more the was slightly one divergent most were the genes, tRNA Among group genes. other than trees Japanese gene rRNA 12S Northern and tRNA that the indicated in genes mtDNA-encoded substitutions the nucleotide on the based trees Phylogenetic strains. inbred 4 and We variation. genetic local distinct stocks local a 8 among Medaka of DNAsequences mitochondrial (mtDNA) whole the and compared distribution geographic large a has and (10–40°C) Medaka ofIntegrated Biosciences,Department The University of Tokyo, Kashiwa, Japan H. Mitani adaptation cold and DNA mitochondrial medaka in repeat tandem of number between relationship The 445 is very hardy and tolerate a wide range of salinities and temperatures and salinities of range wide a tolerate and hardy very is latipes Oryzias Thr gene as reported in humans previously. The number of tandemly repeated 11repeated tandemly of number previously.The humans in reported as gene Gene regulation Oryzias Oryzias somites, diencephalonandbloodat24hourspostfertilization. clustering allowed us to definehierarchical by severaldata groupsexpression ofof grouping genesThe co-expressedbrain). inadult different36hpf, to organs16hpf including of (pool libraries different 2 from activity regulatory transcriptional with genes 900 of cDNA full-length isolated transcription with genes 668 generalclear include DNA to binding 142 also domains, but which comprehensive were and be missed in to precedent only remodelling genome-wide not TF shown screens. was chromatin Weset This then infactors. implicated genes 546 DNA TFs, 1939 includes binding genes of set virtual this of coverage The (ZFishv7). genome zebrafish the map to domains InterPro on based regulators transcriptional of repertoire a We established first in ordertogeneratesituexpressiondataandprovidecDNAsforfunctionalstudies. genes TF zebrafish all of cDNAs full-length of collection a of establishment the is project this of stage specificity, resolution and coherence of annotations and expression patterns. The objective and overall the in situ databases produced so far in many vertebrates are of limited assembly)use ZFishv7 regarding ZFin, in annotation an have TFs of 52% (only small relatively is zebrafish in known TFs of pattern Surprisingly,expression differentiation. the and determination fate cell to proliferation from behaviour cellular of aspects many control and expression gene of regulators and development embryonic of genetic diseases. understanding This is especially and true in networks the case regulatory of transcription factors gene (TFs), which of are critical construction The knowledge of gene regulation both in space and time provides essential information for the Karlsruhe Institutefor Technology (KIT) Institute of Toxicology andGenetics(ITG), Forschungszentrum Karlsruhe, Germany O. Armant, M.März, J.Lampert,L. Yang, M.Ferg, S.Rastegar, U.Strähle Genome wideexpressionanalysisoftranscription factorsinzebrafish embryos Gene regulation 446 517

Posters Posters elements andcognatebindingproteinsinvolvedintestis-specific geneexpression. 1) to express transgenes specifically in the male gonad, 2) to discover new discover to 2) gonad, male the in specifically transgenes express to 1) In conclusion, the short GSDF promoter fragment presented in this study will be a valuable tool to studiesinmammals. some of them are known to be involved in specific gene expression in the Sertoli cells according are conserved within the first 600 bp. These regions contain transcription factors binding motifs, from collected sequences promoter zebrafish,proximal medaka, stickleback,GSDF fugu andthe trout tissues genomesof showed other thatanalysis three complex the comparative DNAAovary. regions in the detectable including barely tissues female the was in detectable was and expression transgene no testis addition, In examined. the in mainly detected was expression Transgenemedaka. male adult the in transgene the of expression the detect to us allowed PCR RT-quantitative the of However,use development. the early the during techniques observation lines transgenic both that macroscopic indicating using GFPdetectable No was site. expression insertion transgene single a harbour observed was transgene gene the (DrGSDF:GFP). of GSDF protein segregation fluorescent the green mendelian Athe of of frame fragment reading producedtwo promoter open the wehave 2kb to upstream a testis, inserted carrying the of lines cell medaka transgenic Sertoli the independent in specifically expression gene drive to In order to determine whether the zebrafish proximal promoter of the GSDF gene was sufficient the orthologousGSDFgenesareexpressedinmalegonadofbothzebrafishandmedaka. that demonstrate we addition, syntheny.In a to belongs gene GSDF the that indicating and species order identity, The tetrapods. in not but orientation of species the genes teleost surrounding in the GSDF genes counterparts are orthologous well conserved found among the and teleost vertebrates in genes GSDF the of analysis phylogenetic a out carried Wehave spermatogonia. Dev Biol, 301:266). This factor increases the proliferation of the primordial germ cells and early (TGF)-β superfamily recently identified in trout Sertoli and Granulosa cells (Sawatari et al., 2007, The Gonadal Soma Derived Factor (GSDF) is a new member of the Transforming Growth Factor Gif-sur-Yvette, France 1 518 A. Gautier fish testis Thezebrafish drivesthe gene of GSDF promoter proximal transgene in specifically expression SCRIBE,INRA,1037 UPR Beaulieu,de Campus Rennes, FRANCE; 447 1 , C.Melin 1 , F. Sohm 2 , J-S.Joly 2 , F. LeGac 1 , J-J.Lareyre 1 2 GIS AMAGENINRA,CNRS Gene regulation cis DNA regulatory DNA and P. Boudinot FinTRIMs: a highly diversified gene family of teleost fish with the hallmarks of antiviral thehallmarks with fish J-P.Levraud ofteleost family gene restriction factors diversified highly a FinTRIMs: Gene regulation involved inimmunityidentifiedoutsidemammals. finTRIMs suggest an important role in fish innate immunity; evolution this would the of make features diversity, them and The that evolution, shuffling. theexon by firstintertwined indicating was TRIMs NLRs and TRIMs (NLR), of receptors closely NOD-like among most found domains are B30.2 finTRIM The to mammals. related in motif recognition viral a within concentrated under positive selection are remarkably congruent in finTRIMs and in mammalian TRIM5alpha, positions The selection. positive strong under evolved finTRIMs zebrafish of domain B30.2 The species. The closest mammalian relatives are trim16 and trim25, but they are not lineage- true orthologs. suggesting subsets different specific diversification revealed events. anddisplaying Accordingly, analysis the number phylogenetic chromosomes, of fintrim A ondifferent genes varies greatly polymorphism. among inclusters fish strong transcripts. distributed diversity 84 genes striking asvirus-induced a harbors with genome finTRIMs, trout zebrafish of The domain. B30.2 a inrainbow include variants long the first length; variable of C-termini and regions identified RING/B-box identical nearly of formed are finTRIMs, FinTRIMs called teleosts, TRIMs of subfamily new in large a describe We selection. positive and expansion gene through strong selective pressures on the defense system, and Virusesexert primates. antiviral many to TRIMs HIV to resistance show confers which protein signs TRIM5alpha the of being diversification example known best the infection; viral against immunity intrinsic including processes cellular various in involved are family protein (TRIM) motif tripartite the of members the mammals, In Développement del’Immunité, InstitutPasteur, CNRSURA2578, Paris, France 1 iooi e Imnlge oéuars INRA, Moléculaires, Immunologie et Virologie 2 , L. van der Aa der van L. 1 1 , M. YahmiM. , 1 , E. Lauret E. , 1

, V., Briolat oye-oa, France; Jouy-en-Josas, 2 , P., Herbomel 448 2 , A. Benmansour A. , 2 arpae et Macrophages 519 1 ,

Posters Posters 520 membrane protein, groupof in the basement (hmcn1). novel potential a wasidentified encodes gene and Athird mutants morphology. zebrafish fin proper for fundamental is This interact with Fras1 and maintain the stability of the protein complex in the basement membrane. membrane proteins to stabilize dermis-epidermis structure. As in mice, zebrafish Frem2 seems to lung of consequence or renal dysfunctions. a During development Fras1 and Frem2 as work together with other die basement newborns the of 45% around and syndactyly cryptophthalmos, present patients The syndrome. human, Fraser as known disorder multisystem in recessive a to related genes, are those of orthologues The proteins. genesas membrane two basement related structurally haveidentified we cloning, positional Applying fins. We have studied a group of non-allelic zebrafish mutants with blistering in the skin of embryonic Institute forDevelopmental Biology, Cologne University, Germany N.M. Feitosa, T. Carney, J.Kluger, M. Hammerschmidt The roleofhemicentininfindevelopment been showntointeractdirectlywithFras/Fremproteins. Hmcn1 indicates that it can bind to nidogens and other basement membrane proteins that have possibility thattheproteinsinteractinanindirectmanner. Themultidomainproteinstructureof Fras1 protein stability within the basement membrane. However, this result does not exclude the fold. fin while cells, epidermal in expressed Zebrafish orthologue. two Vertebrateshave development. of stages same the around 449 nagel nagel (hmcn1) mutants, mutants analysis indicates that in contrast to Frem2, Hmcn1 is not required for required not is Hmcn1 Frem2, to contrast in that indicates analysis mutants frem2 frem2 (blasen) and is primarily expressed in mesenchymal cells of the of cells mesenchymal in expressed primarily is hmcn2 fras1 (pinfin) have similar blister formation fras1, ht encode that frem2 hemicentin1 is hmcn1 M. Andersson Lendahl500 M. Anchelin 416 C. Anastasaki 388 R. Amodeo 312 J.F. Amatruda 75 V. Amato 284 J. Amack 46 Y. Alvarez 289,352448 M. Alvarez 263 A. Alunni 200 J.P. Almeida-King 445 C. Almås 465 K. Alitalo 315 D.W. Ali 181 J.M. Alfaro 366,473 M. Alders 90 F. Alcaraz-Ruiz 337 F. Alcaraz-Pérez 325,416 M. Alcalay 165 R.C. Albertson 428 A. Akalin 451 T. Aizu 442 H. Aizawa 101 N. Aghaallaei 96 D. Aggad 334 M. Agetsuma 101 T. Adachi 199 L. Abrami 182 S. Abke 100,247 S. Abdelilah-Seyfried 299 L. Abbas 218,35 A Authors Index B. Ayari 363 R. Avellino 438 N.S. Asuri 262 O. Astudillo-Fernandez 314 R. Ashworth 178 K.B. Artinger 511 A.B. Arrenberg 273 E. Aronica 356 J. Arnout 345 H. Arnardóttir 296 O. Armant 517 A. Ariza 316 A. Arigony 498,505 F. Argenton 251,38396 R. Arévalo 285 M. Arese 304 S. Ares 490 A. Archer 500 C. Araya 480 L. Araujo-Bazan 372 L.A. Appelbaum 224 B. Appel 261 T Aoki 101 R. Aoki 54 A. Anusha 502 V. Anelli 84,411 M. Andreazzoli 203 M. Andrade 215 F. Baas356 S. Buraschi301, 407 B M. Behra108 H. Begthel58 T. Becker 276 C.G. Becker276 L. Beccari485,6 S. Baxendale466 A. Baumbach428 G. Basso420 C. Barton146 P.G. Barth356 G. Barsacchi203 E. Barriga263 A. Barrallo-Gimeno222 S. Barlati378,390 I. Barinaga-Rementeria M. Barilari203 S. Banks384 T.U. Banisch4 S. Banfi438 L. Bally-Cuif383 J. Balciuniene63 D. Balciunas24,63 K. Balasubramanian443 J. Bakkers409 B. Bajoghli96 T. Bailey106 H. Baier273,4674715 B. Bahn462 C.P. Bagowski409,410 Y.K. Bae254 A.P. Badrock57 G. Badaracco376 Ramirez 403 521

Authors Authors 522 N. Bolli347 M.R. Bogo508 T. Boehm 96,32332 J. Blechman258 B. R.Blazar431 B. Blanco292 P. Blader497 A. Bisazza294 A. Biondi313 A.C. Binot173 H. Bielen260 I.H. Bianco234 R. Bhat362 N. Bhat104 S. Bessling513 J. Bessa451 J.Y. Bertrand92 J. Bertrand97 J.G. Berthet227 C. Berndt227 J.N. Berman408 S. Berger 393 J. Berger 452 I.M. Berger 87 G. Bergamin 396 V. Berg 465 A. Benmansour519 A. Benini378,390 F. Benato168 M. Beltrame304,312 H.G. Belting156 A I.Belo410 D. Beis308 R.H. Brookfield469 J.D. Brook437 D. BroockRosemberg 498,505 C. Broesamle287 J. Brocher360 V. Briolat519 S. Bretaud382 R. Bresciani202,390 E. Bresciani340 T. Brand307 M. Brand47,612292425 P. Braghetta171 L. Braeutigam227 D.F. Bradley421 A. Bozzato390 B. Boyer295 E.R. Boyd408 P. Bovolenta438,4856 P. Bouvet263 F. Bourrat200 D. Bournele308 S. Boué432 P. Boudinot519 F.L. Boss90 G. Borsani202,378390 P. Bovolenta389 F. Bontems43 J. Bonkowsky225 C.D. Bonan498,505 P. Bonaldo171 H. Bolz375 F. Bollig60 508 C. Campos100, 303 S. Campanaro25 7 T. Camarata322 L. Calvarini390 D. Calebiro168 Y. Cai314 C L. Byrnes205 M.F. Bustamante272 A.S. Busta368,380 F. Bussolino304 J. Bussmann90 K. Bushby371 E. Busch-Nentwich454 C.E. Burns95 S.M. Burgess 69, 108 C. Burckly 397 S. Buraschi301,407 B.S. Budde356 A.J. Buckle231 N. Buchan481 E. Bubenshchikova446 E. Brustein367 C. Brusegan489 W.R.A.Brown 469 S.B. Brown327 K.H. Brown66 J. Brown37 K. Brown421 A. Brouwers239 C. Brösamle394 Authors Index W-T. Chen206 J.K. Chen65,352 GD. Chen166 A. Chen95 R. Chauhan70 J. Chau428 A. Chatzopoulou167 P. Chapouton462 J. Chang213 G. Chandrasekar500 G. L.Chandler75 N. Champagne367,395 A. Chakraborty510 J.F. Chai164 S. Cermenati312 J. Cerda147 G.A. Cerda259 C.C. Ceol7 M.L. Cayuela325,416 F. Cavodeassi488 I. Castanon100 F. Casares451 R. Carvalho72 S. Carrella438 S. Carra340 T.J. Carney82 T. CarneiroBrandãoPereira G. Cappellano507 M.C. Capogrossi313 L. Caneparo39 S. Candel338 M.L. Cancela267 Authors Index 498, 505, 508 S. Colanesi402 T.S. Coe 228 D.E. Clouthier509 B. Clissi479 H. Clevers58,373 A. Clement209 A.E. Clatworthy401 J.D.W. Clarke472 M. Clark71 E. Cisneros485,6 S. Cimbro304 F. Cignarella390 F. Ciccarelli411 C. Cianciolo425 H.N. Chuang149 C.Y. Chu166 E.L. Christie57 W. Chow445,456 S. Choorapoikayil177,406 E. Choi74 A. Chitnis176 C-B. Chien266,321482 A. Chien336 T. Chico306 N. Chiarelli378 N. Chi92 G. Chesi389 W.O. Cheong164 H.Y. Cheng149 C.H. Cheng166 Y-Y. Chen20,4 Y-H. Chen370 Y.C. Chen166 F. d’AddadiFagagna 361 M. Dadda294 S. Da’as330, 1 D K.J. Curtin289 P.D. Currie393,398 E. Cuppen58 V.T. Cunliffe 231 W. Cui278 Z. Csenki148 C. Cruz160,163 G. Crump5 P. Crosier80,94336 K. Crosier80,94336 R. Creton223 C.L. Crespo479 B.D. Crawford468 A.D. Crawford191 P. Coutinho514 D. Cottell289 F. Cotelli304 S. Costagliola207 G. Cossu487 P. Corti300 R.A. Cornell104 I. Conte438,4856 M. Connolly428 S. Confalonieri340 N. Conceição267 M. Colombi378 J.E. Collins445 523

Authors Authors 524 J.E. Dick347 P. Devanna313 C. Detzer309 N. Detry192 G. DeSena301,407 G. DeRienzo363 M.L. Dequeant52 J. denHertog177,406 B. Demarest83 A. Delogu226 G. DelMoro420 A. DellaPuppa257 I. Della489 L. DelGiacco359 M. delCarmenRamos366 F. DelBene32,927475 E. delaCalle-Mustienes451 J.L.O. deJong95 G. Deflorian84 T. deFilippis168 I.F.M. deCoo374 E. deBruijn58 A. Deepa365 M. Debiais-Thibaud265 D. Dormann377 A.J. Davidson60 A. Davidson195 V.M. Darras191 C. Danesin255 C. Dambly-Chaudière484 M.J. Dallman327 F. Dai169 F. Dafhnis-Calas469 M. Ekker265,40 4 O. Ek210 J.S. Eisen230 B.F. Eames56 E R. DutraDias498 S. Duga359 S. Dudczig244 H.J. Duckers90, 315 D. Duboule208 W. Driever41,15621024 P. Drapeau243,9367395 J.J. Dowling368,380 X. Doss298 R. Dosch12,43 B. D’Orsi203 D. Dormann377 J.R. Dorin333 C. Dooley400 S. Donaldson444,5 A. Domenichini251 J. T. Dobson408 G. Djordjevic60 M. Distel64 R. Dirks72 Dipietromaria A. 162 H. Dill360 S. Diks239 P.P. DiFiore340 C.Q. Diep23,60 D. Diekhoff326 A. Fernández451 M. Ferg 517 F. Fenyes454 M. Fenske470 X. Feng169 H.A. Feldman95 E.L. Feldman368,380 B. Feldman37 N. Feitosa-Martins82 N.M. Feitosa520 M. Featherstone502 A. Faucherre107,17271 C. Fassier103 U. Fascio313 A. Faro58 L. Facchin102 F C. Eugster492 R. Ethier193 A.L. Estep388 T. Erickson232 I. Erhard415 D. Epting302 K.E. Epley85 C. Englert60 A. Emelyanov419,443 S. Elworthy164 S. Ellingsen172 P. Elks329 S.C. Ekker70 406 Authors Index C. Fritegotto168 D. Freudenreich61,229244 K. Freson345 D.V. French204,232 C.R. French232 J.L. Freeman6 J.K. Frazer76 S.E. Fraser39 B. Franco389 D. Fraher428 N.S. Foulkes49 A.E. Fortunato208 A.M. Forrester341 M. Folgueira472 E. Foglia304 K.E. Fogarty8 K. Fluiter356 G. ParkerFlowers185,6 M. V. Flores94 P. Flicek67 A. Fjose172 S. Fisher512 U. Fischer360 T.S. Fiona502 E. Finch163 A. Filippi240,257 A.L. Filby228 H.A. Field57 F.F. Ferre7 M.I. Ferrante81 A. Ferrando331 C. Fernández432 Authors Index 256 A. Ghilardi359 G. Gestri203,219 M. Gesemann280,291 A. Germanà250,284 M. Gering311,3448503 S. Gerety459 A.S. Georgiou 231 F. Geng358 R. Geisler400,45 F. Geier41 J. Gehrig68 M. Geffarth256 N. Gebhart252 A. Gautier518 G. Gaudenzi507 H. Gaude397 L.G. Garraway7 G. Gariano161 M. Garcia-Lecea 447 Z. Garavito-Aguilar324 J. Ganz229,244256 M. Gallagher51 A. Gal375 M. Gajewski298 G T. Furukawa281 M. Fürthauer100 A. Fujiyama442 K. Fujita214 N. Fujimori235 C.Y. Fu201 M. Grealy205 H. Grandel229,256 M. Graf360 C. Grabher346 V. Gouriev58 D.A. Goulding81 K.L. Gould209 K. Goudevenou151 M. Goudarzi 4 D. Gotti202 M. Goto101 Y. Gothilf49 B. Gorsi306 L. Gordon242 J.M. GonzálezRosa316 R. Gonzalez-Quevedo98 M. González-Gaitán100,303 S. Gonzalez372 K. Gonzalez63 M. Gonzalez-Gaitan182,247 Z. Gong419 J-L. Gómez-Skarmeta316 E. Glynn52 T. J.Glass431 B. Giros103 A.J. Giraldez89,184 J. Gilthorpe226 P.C. Gilligan45 M.J. Gilchrist67 J.G. Gilbert444 G. Gibon282 E. Gibbs380 J. Ghosh163 525

Authors Authors 526 Y. Han154 H.W. Han 166 K.L. Hammond105 M. Hammerschmidt212 M.E. Halpern102 M.C. Halloran262 T.E. Hall 398 C. Hall80,94336 L.A. Hale230 J.L. Hakkesteeg145 M.A. Haendel73 Y. Hadzhiev68 C. Haass362,3779 H A. Gutnick258 P. Gut216 T. Gupta43 F. Günthner307 D. Gunther46 J. Guinea372 I. Guerrero-Garduño190 M.C. Guerrera250 M. Guarienti161 A. Guarda376 L. Guajardo263 D.J. Grunwald46 B. Grotek435 J.M. Gross268 Y. Grinblat238 M. Grimaldi195 E. Griffiths445 J. Hescheler298 R. Herpers315 J-M Hermel200 C. Herd454 P. Herbomel334 R.C. Hennekam90 A.T.M. Hendrickx 374 C.P. Heisenberg 182, 481 C-D Heidecke409 A. Hegyi148 Z. Hegedus328 L.W. Heaton230 J.K. Heath452 S. He72 J. Hazan103 T.A. Hawkins472 G. Hausladen383 G. Hauptmann227,241 M. Haug280 A.K. Hatzopoulos320 G. Hattem52 K. Hatta214 D. Hassel319 H. Hashimoto446 M.R. Harrison231 W.A. Harris496 M. Harris40 G. Harms476 A.P. Haramis373 S. Hans24,612425 R.D. Hannan217 R.I. Handin60 N. Hanahara235 K-L. Huang283 F-L. Huang245 R. Hua443 J. Hu320 D. Hu83 J-C. Hsu503 K.M. Hsiao150 A. Hruscha377,9 K. Howe444,5456 Y.H. Houvras7 C. Houart103 W. Horsley76 J. Horsfield384 D.S. Horner507 J.H. Horne460 S-K.Hong 37 J. Holzschuh413 A. Holmgren227 E. Holmes437 N. Holder208 M. Holder495 B.M. Hogan315 R. Hoffmans53 S. Hochmann229 A. Hirmer375 H. Hirata278 J. Hillmer248 S. Higashijima101 M. Hibi254 A. Heuze200 E.C. Hett401 I. Hess326 B. Hesl383 Authors Index J.C. IzpisúaBelmonte432 E. Izaurralde504 N. Iwanami332 E.G. Ivashkin153 S. Isogai317 H. Ishizaki402 E.Y. Isacoff9 K. Inoue439 P.W. Ingham164 A. Indrieri389 F. Imai189 J. Iklé509 T. Ikenaga 252 A. Iervolino203 G. Iaffaldano313 I D.R. Hyde106 S.P. L.Hwang166 I.G. Huttner398 J. Hutt103 S.A. Hutchinson83 M.R. Huska215 A.F. Hurlstone405 E. Huntzinger504 D.T. Hung401 C-C. Hung245 S. Hundt310 N.A. Hukriede60 J. Huisken307 M. Hughes403 T. Hubbard444,5456 Y-Y. Huang280 Authors Index H. Kagoshima44 2 B. Kagermeier-Schenk 435 E. Kabashi367,39 5 K P.R. Jusuf496 S. Just87 M. Jurynec46 B. Jungblut309,310 F. Jülicher493 J.L. Juárez-Morales246 E. Jongen374 P. Jones151 M.E. Jones405 J-S. Joly200,518 A.B. Johnston347 I.Þ. Jóhannesdóttir296 L. Jing242 S. Jia169 J.R. Jessen159 K-H. Jeong74,41 J-Y. Jeong74,41 H-Y. Jeong74 L. Jensen314 E. Janssens236 J.J. Jane-Valbuena 7 F. Jamen200 C. Jainok240 R. Jagasia462 I.J. Jackson402 J S. Kitambi501 B.C. Kirchmaier 307 M.L. Kirby86 S. Kinoshita194,506 M. Kinoshita446 E. Kimura317 C.B. Kimmel56 H.R. Kim175 R. Kettleborough454 A. Ketley437 U. Kern225 K. Kern302 B.N. Kennedy289 N. Kenmochi510 R.N. Kelsh174 P. Keller, 479 D. Keefe67 K.D. Kaya457 K. Kawakami361 H.A. Katus319 E. Kastenhuber225 S.C. Kassen106 J. Kaslin61,22924425 P. Kasher356 K. Karlsson296 M. Karali438 M. Kapsimali274 J.P. Kanki347 S. Kani254 E. Kalmar68 M. Kai481 F-B. Kai408 E. Kague513 527

Authors Authors 528 A. Kumichel52 J. Kuja-Panula253 K.M. Kuerner264 T. Kudoh 458 A. Kudo349 G. Kronnie420 J. Kroll302 A. Krol52 V. Kroehne244 C. Krivcevska178 M. Kraussling152 M. Kraeussling170 B.Y. Kovacs148 Z. Kostun193,58 V. Korzh 447 J. Korving58 M. Köppen38 W.J.A. Koopmans62 O. Konu457 I. Kondrychyn447 T. Kondoh208 S. Kondo196 M. Konantz400 Y. Kohara442 C.H. V. Koh419 R.W. Koester362 K. Kobayashi461 E. Knust290 L.D. Knogler249 E.W. Knapik222 N. Klüver449 D.A. Kleinjan514 E. Klee70 M.Y. Law266 S.H. Laval371 G. Lauter241 E. Lauret519 R. Laurà284 C. Laufer319 P. Lau311 J-J. Lareyre518 M. Lardelli353,369 M. Lapointe367 M. Langworthy261 F. Langellotto220 L. Lanfrancone361 E. Landi203 J. Lampert517 J.R. Lamb327 E. Yi NiLam94 M.K.Lalwani 70 J. Laisney78,415 G.K. Laird445 M. Lahne178 A.K. Lagendijk409 M. Lacovich313 K. Lachani67 M. Lachance286 L H-J. Kwon104 L. Kwok511 K. Kwan482 J.Y. Kuwada278 V. Kuscha276 C. Lillesaar383 E.G. Lightman231 G. Lieschke393 G.J. Lieschke57 D. Liedtke78,415 U. Liebel68 M. Liao367 B-K. Liao491 J. Liang69 S. Li422 C. Li422 K. Lewis246,259 J. Lewis495 J-P. Levraud334 G. Levkowitz258 C.Levin 37 L. Leung474 M. Lerch 79 S.Y. Ler515 A. Lepier462 F. LeGac518 K.K Lefler148 Y. Lee1428 O. Lee74,41 J. Lee513 H-O. Lee74,41 H. Lee26,744142 H-C. Lee201,6 C. Lee6,20142 C.H. Lee51 E.E. LeClair109 E. Lechman347 N.D. Lawson88 Authors Index K. Lunde156 T.C. Lund 431 A. Lumsden226 J. Lu422 L.A. Lowery213 C.A. Loynes342 M. Lowe403 S. Low278 S. Louwette345 R. Louren494 C. LoraLamia313 H. Lopez-Schier107 M.A. López-Muñoz325 M. Lopez212 S. Lopes494 A.T. Look347 E.E. Locke83 H. Lochmüller371 D.M. Lloyd444 Y. Liu159 M. Liu50 J.J. Liu355 F. Liu306 J.A. Lister402 B. Link270 B. Linder360 R.E. Lindeman183 Y-Y. Lin 385 S. Lin422 L. Lin395 C.Y. Lin206 Y.M.A. Lim355 C.H. Lillig227 Authors Index S.S. Markmiller57 K. Mark401 W.M. Marin224 R. Marco Ferreres438, 485 R. Marais388 M. Marai429 E. Maquet207 M. Manzoni202 I. Manfroid192 E.M. Mandelkow362 E. Manalo269 L. Malvaux440 N. Malissovas387 J. Malicki281 S. Malhotra70 E. Maldonado190 C. Maios367 J. Maini70 A. Magnabosco399 R. Madelaine497 D. Macmillan333 A. Machate47 R. MacDonald265 A. MacDonald336 X. Ma306 D. Ma60 M J.L. Lyche 465 J. Lyautey 43 M. Luz290 T. Luo187 N. Mercader 316 A. Meng154 D.W. Melton412 C. Melin518 S.H. Meijsing471 A.H. Meijer328 S. Meierjohann78,170415 S. Megason477 N. Meeker421 B. Meder87,319 T. Meckel62 N. Meani165 K. McMahon311 S. McLoughlin314,448 C.E. McDeed193 M. Mazel334 E.A. Mayhall95 A. Mavropoulos212 C.M. Maurer288 L. Matthews453 M. Matsuda176 S. Mathavan360 I. Masai54,1889 K. Maruyama349 E. Martin-Blanco158 P. Martin151 E.D. Martin205 J.A. Martial440 J. Marshall330 M. Marsal158 I.J. Marques 409,410 G.M. Maro224 F. Marlow43 529

Authors Authors 530 L. Morelli493 L. Moons236 L. Montoliu451 E. Monti202,418 R. Monteiro335 C. Monteccucco399 G. Montalbano250,284 P.K. Moly214 R. Molteni479 A. Molina263 S. Moleri312 R. Moessner41 M. Moennich384 N. Modena257 A. Mode501 T. Mochizuki188 S. Mitra343 T.J. Mitchison346 D.L. Mitchell351 H. Mitani516 Y. Mishima89 M. Mione84 A. Miñán451 Y. Minakuchi442 M. Milanetto171 E.M. Mignot224 L. Micale418 M. Mia401 M. Gajewski298 G. Messina487 D. Messerschmidt41 J. Meseguer338 G. Merla418 J.W. Nelson 155 C.M. Nelson106 A.C. Nelson441 E. Negrisoli257 F. Naye192,210 K. Naruse442 S. Narawane172 S. Naranjo451 Y. Namavar356 S. Nakayama214 R. Nakayama101 T. Nakata 199 A. Nagiel277 D. Nagelberg 63 N A. Muto9,471 V. Muñoz263 M.C. Mullins43 J.S. Müller371 M. Muller212 V. Mulero325,338 F. Mueller68 M. Moyano488 P.M. Mourrain224 P. Motte173 M.L. Mostacciuolo396 E. Moro381,396 R.H. Morley67 A. Moriyama349 M.A. Moriarty205 M. Moreno325,416 L. O’Brien195, 8 E. Ober57 A.C. Oates464 O M.K. Nyholm238 C. Nüsslein-Volhard 40,40 P. Nürnberg 356 R.J. Nuckels268 M. Nowak47 R. Nourizadeh-Lillabadi465 A. Norris427 S. Nornes353,369 C. Norden270,474 R. Noche478 S. Noble404 O. Nivelles440 C. Nitsche409 R.W. Nipper76 P. Niethammer346 R. Niemann298 S. Nicoli301 G. Nica212 M.T. Nguyen275 H.B.G. Ng463 A. Ng268 M. Newman353,369 J. Neumann75 S.C. Neuhauss291 S.C.F. Neuhauss280,8 D.S. Neuberg 330 K.A. Nembhard86 Authors Index L.I. Partecke79 A.C. Parslow57 S. Park322 S. Parinov419,443 J. Paridaen239 V. Parente429 R. Pardi479 D. Paquet362,3779 P. Panula253,386 P. Pantazis39 V. Pandey70 H. Pan149 G. Paganini202 P R. Ozturk457 H. Ozdag457 X. Ouyang65 H. Otsuna482 A. Oswald490 R. Opitz207 Y. Ono194,506 F. Ono252 D. Onichtchouk41,156 Y. Omori281 K. Okuda80 H. Okamoto54,427 K. Ohishi442 S. Ohata54 M. Ogueta285 S. Oehlers80 J. O’Connor352 Authors Index B. Polok41,156 B.K. Polok272 G. Polo396 F. Pistollato420 A. Pistocchi359 T. Piotrowski 275 S. Piloto48 D.B. Pilgrim204,232 L. Piffer294 T. Pietri269,27 R. Philp45 J.B. Phillips292 F. Pezzimenti84,14 M. Petron399 T. Peterkin305 M. Pesce313 L. Persani168 S.L. Perkins421 E. Peris366 T. CarneiroBrandãoPereira M. Peppelenbosch436 H-C. Peng370 H. Pendeville440 F. Pelegri183 B. Peers173,192210 E.M. Payne347 E.E. Patton333, 427 S.A. Patten181 A. Patowary70 R. Patient306,335 M.R. Passos-Bueno513 E. Pashos512 498, 505 K.A. Rauen388 S. Rastegar517 E. Rampazzo420 J.L. Ramos-Balderas190 D. Rambaldi411 A. Ramanathan443 B. Rajasekaran493 R Y. Quiroz212 H.N.B. Quach443 Q R. Purkanti70 T. Purea336 J. Pujol-Marti271 E. Pugach95 M. Priyadarshini391 F. Priller215 M. Presta301,407 A. Prades366 O. Pozzoli429 M. Poyatos366 J. Postlethwaite71 A. Porteros285,488 M. Porte216 I. Porreca221 P. Porazzi168 A.N. Popper69 K. Pooja45 G. Pompilio429 531

Authors Authors 532 I. Rodriguez-Martin372 F. Rodrigues174 F. J.Roca337 A.L. Robertson342 A. Roberts246 M. Ritelli378 A. Rissone304 J. Ripoll366 B.B. Riley104 M. Rigoni399 Y. Rifat57 E. PachecoRico498,505 Y. Richaud432 J. Richardson414 N.L. Reynolds333 A.L. Reynolds289 A.E. Reyes499 M. Reuter470 S. Renshaw329 S.L. Renninger291 P. Renner449 J. Renn211 P. Rengtved-Lundegaard402 G. Renaud69 M. Reischl68 B. Reimholz444,5456 M.M. Reimer427 L.M. Reaume341 E. Raz4 F. Raycroft106 M. Raya432 Á. Raya432 H. Rauvala253 B. Sapetto-Rebow289,448 B. Santoso92 A. Santos-Ledo285,488 M.M. Santoro91 C. Santoriello84,361 L. Sangiorgio 304 N.A. Sanek238 K. Sampath443 R. Salomon397 S.A. Sakowski368 H. Sakamoto439 L. Saint-Amant269,286 A. Sachinidis298 F. Sablitzky151 S L. Rygier 331 J. Ryan 269,286 J.M. Ruiz485 F. Ruf-Zamojski477 L. Rudner421 J. Rowlinson344 M. Roussigne234 W. Rottbauer87,319 I. Rothenaigner462 I. Roszko159 O. Rossetto399 F. Rosa295 F.M. Rosa274 I. Rønnestad172 B.L. Roman300 D. Roellig52 D.F. Schorderet272 H.B. Schönthaler288 P. Schoonheim167,273 S. Scholz449 S. Scholpp103 K. Scholich309 S. Schneider-Maunoury 103, E. Schnapp487 T.S. Schmidt62 B. Schmid362,3779 H. Schirmer505 S. Schindler68 H. Schimer508 T. Schilling48 V. Schiller470 A. Schiavinato171 F. Schepis489 A. Schepis155 K. Scheffler449 M. Schartl152,1704145 C. Schäfers470 K. Schaefer287 M.J.M. Schaaf167 V. Scaria70 E. Sawatari199 A. Sawada483 S. Saunier397 L. Saúde494 M. Sartori239 S. Sarmah222 T.D. Sargent 187 179 Authors Index T. Sýker 470 M.K. Sýffker228 C. Stewart-Swift357 C. Standley88 N. Soussi-Yanicostas 162, 363 J.U. Skåre465 F.C. Simões305 A.F. Siekmann88 T. Shiraki 101 M. Shinya446 T. Shin-I442 L. Shine289,448 T. Shimizu254 I.A. Shestopalov65 D. Sheng355 S. Shaw401 M.P. Sepulcre337,8 D. Sepich159 C.A. Semple333 R. Selleck195 R. Seki199 H.K. Sehra445 E.K. Scott9 T. Schwerte307 J. Schweitzer210 H. Schwarz 400 S. Schulte-Merker90 D. Schul476 P. Schu161 C. Schroter42 M. Schorpp326,332 Authors Index J. Tolar 431 C. Tobia 161,30407 N. Tiso 168,202396 J. Timmer 41 B. Tien Poll-The356 R. Thummel106 E. Thoma170 R. Thambyrajah348 M. Teucke 377,9 J. Tena 451 N. Temperley 402 E.M Teh 330 J-M. Tee 239,436 V. Taylor 41 K. Taylor 402 H.B. Taylor 327 A. Taylor 238 O. Tassy 52 Y. Taniguchi 78 M. Tanaka 461 H. Tanaka 430 K. Tanabe 254 X. Tan 157 W.S. Talbot 155 Y. Takeda 439 S. Takeda 78 M. Takamiya 23,59 M. Takahoko 101 M. Tada 458 T.L. Tabone 57 N. Tabaneras 486 T K. Uesugi214 T. Uechi510 U C.R.Tyler 228 M. Tyers 412 E. Turola 489 L.T. Turner 7 K. Turner 472 J. Tuimala 391 B. Tucker 353 L.C. Tu 150 S. Tsuruoka-Kinoshita 54 M. Tsang 203 H-J. Tsai 201,6 A.J. Trotter 217 L. Trinh 477 N.S. Trede 76, 83, 421 D. Traver 92,7 D. Trauner 293 A. Toyoda 442 D. Tosh 174 C. Torroja 454,5 J. Torres-Vázquez 297 M. Torres 316 J. Torrance 444,5456 J. Torrace 455 H. Torihara 510 J. Topczewski 12,109185 J.M. Topczewska 185,6 J. Tong 154 186, 322 533

Authors Authors 534 J.A. Vega 250,284 J. Veerkamp 299 G. Vazza 396 G. Vatine 49 M. Varga 143,426 L.Varadi 148 P. vanTijn 354,436 C. vanRooijen354 T. Van Raay483 J. vanNoort62 M. vanLohuizen373 V. vanHeyningen514 C. Vangeet 345 F. vanEeden454 P. vanDuijn177,406 G. vanderGoot182 L. vander Aa 519 Y.U. vandervelden373 B.J.C. vandenBosch374 T. vanBoxtel357 F. vanBebber377 D. Vallone 49 E. Vaccari 257 V R.A. Uribe268 B. Urbanyi148 J. Urban252 A. Urasaki317 A.U. Uong7 J.A. Ulloa318 S. Uji199 G. Weidinger 180,43 5 J. Wegner 292 B. Weger 59 M. Watanabe 196 S. Watabe 194,506 A.J. Waskiewicz 232 P. Washbourne 269,27 E. Warp 293,475 F.C. Wardle 441 X.G. Wang 164 W-D. Wang 320 N. Wang 422 M. Wang 365 G.W. Wang 224 C.N. Walpita 191 S. Walmsley 329 N. Waghorne 314,352 H. Wada 54 W M. Voz 173,210 E. Voronezhskaya 153 V. Von Berg 192 D. Volpin 171 D.H. Vlecken409 E. Villa 489 A. Vettori 396 C. Vesque 179,397 P. Vernier 383 H. Verkade 57 M.I. Vera 263 P. Vella 313 C. Wyart 467,475 Y-T. Wu 350 C.C. Wu 149 B.K. Wu 166 G.K. Wong 474 T. Wolfsberg 69 S.A. Wolfe 8 N. Wittkopp 504 C. Wittevrongel 345 J. Wittbrodt 479 M. Witte 90 V. Wittamer 30,97 J. Winkler 298 C. Winkler 211,364502 R. Wingert 195 S. Wilton 393 S.W. Wilson 426 L. Wilson 369 C. Wilson 143 T. Wilm 483 B. Willems 211 D. G.Wilkinson 459 B. Wilde 415 J. Widenhain 412 A. Wichmann 470 M.K.B. Whyte342 T.T. Whitfield105,21835 M. Westerfield 292 B. Wendik 156,302 F.U. Weiss 409 C. Weiler 504 Authors Index R.S. Yu 47 S. Young 270 R. Young 426 J. Yost 46 A. Yoshizawa 189 Y. Yoshimura 235 T.Y. Yokogawa 224 C. Yin 483 L. Yieh 268 D. Yelon 324 Y.J. Yeh 151 C. Yanicostas 233,434 Z. Yang 422 S. Yang 154 P-H. Yang 206 L. Yang 517 H-J. Yang 206 J. Yan 206 T. Yamamoto 21 4 M. Yamaguchi 188,235 M. Yahmi 519 Y Y. Xu157,18 P. Xu154 S. Xin422 X Authors Index T. Zygmunt 297 N. Zoppi378 L.Z. Zon26 L.I. Zon95 D. Zizioli161 D. Zivkovic239 L. Ziv471 A. Zink413 Y. Zhou95 W. Zhou278 H. Zhong422 X. Zheng154 Y. Zhao422 X. Zhao253 Z. Zhang365 Y. Zhang157 P.J. Zhang157 B. Zhang2 H. Zhan423 Z. Zeng423 A. Zaucker68,14 A. Zakrzewska 72 Z 535

Authors First published june 2009 Arti Grafiche Ramberti, Rimini, Italy Organizing Committee Marina Mione - Milan, Italy Karuna Sampath - Singapore, Singapore [email protected]

Francesco Argenton - Padua, Italy Massimo Santoro - Turin, Italy [email protected]

Organizing Secretariat

Adria Congrex srl and Meetitaly Via Sassonia, 30 - 47900 Rimini - Italy [email protected]

Congress Venue Palazzo dei Congressi Piazza John Kennedy, 1 00144 Rome - Italy

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