Report 2003-2008 Report 2003-2008 Editors

Report 2003-2008 Report 2003-2008 Editors:

The Board of Directors

Interdisciplinary Center for Neurosciences (IZN) University of Heidelberg Im Neuenheimer Feld 364 D-69120 Heidelberg

www.izn.uni-heidelberg.de

Concept, Coordination and Editorial Office:

Dipl.-Biol. Ute Volbehr Susanne Kilian M.A. Cover illustration by Simon Wiegert, Neurobiology, IZN: Dr. Otto Bräunling Depth-coded maximum z-projection of hippocampal CA1- neurons expressing Dronpa-labelled ERK1. The colour-spectrum from violet to red was used to artificially colour-code the position Production and Layout: of neurons in z-direction. Rolf Nonnenmacher, Dipl. Designer (FH) Image Sources: Sven Weimann Images were provided by the IZN Investigators and the Heidelberg University Hospital Media Center, the University of Heidelberg, the German Cancer Research Institute, Photolab/European Printed by: Molecular Biology Laboratory, Max Planck Institute for Medical Research, and Central Institute for Mental Health Mannheim. CITY-DRUCK HEIDELBERG

2 Table of Contents

Table of Contents

Boards 5 Thomas Kuner 93 Siegfried Mense 96 Research groups / IZN-Investigators 6 Andreas Meyer-Lindenberg 99 Concept and strucure of the IZN 9 Hannah Monyer 101 Appointments 14 Ulrike Müller 104 Christof Niehrs 107 Awards 16 Sabina Pauen 110 IZN Funding and grant giving bodies 18 Gabriele Elisabeth Pollerberg 113 Gudrun Rappold 116 Guest scientists 19 André Rupp 119 Collaborations 21 Martin Schmelz 122 Research Groups 35 Gerhard Schratt 124 Detlev Arendt 36 Christoph Schuster 127 Hilmar Bading 39 Günther Schütz 130 Dusan Bartsch 42 Markus Schwaninger 134 Martin Bohus 45 Peter H. Seeburg 136 Francesca Ciccolini 48 Horst Simon 137 Andreas von Deimling 51 Thomas Söllner 140 Winfried Denk 54 Rainer Spanagel 143 Andreas Draguhn 55 Hartwig Spors 147 Uwe Ernsberger 58 Rolf Sprengel 150 Thomas Euler 61 Kerry Tucker 153 Christian Fiebach 64 Klaus Unsicker 156 Herta Flor 67 Wolfgang Wick 159 Stephan Frings 70 Otmar D. Wiestler 162 Peter Gass 73 Joachim Wittbrodt 164 Darren Gilmour 76 Gabriel Wittum 167 Thomas Holstein 78 Veit Witzemann 170 Stefan Kins 81 Teaching 173 Joachim Kirsch 84 Seminars and Symposia 201 Georg Köhr 87 Rohini Kuner 90 Map of Campus 232

3 4 Boards

Boards

Board of Directors Scientific Advisory Board

Prof. Dr. Hilmar Bading (Acting Director) Prof. Dr. Adriano Aguzzi, MD, Zürich Prof. Dr. Winfried Denk Prof. Yehezkel Ben-Ari, MD, PhD, Marseille Prof. Dr. Andreas Draguhn Prof. Dr. Anders Björklund, Lund Prof. Dr. Herta Flor Prof. Dr. Magdalena Götz, München Prof. Dr. Stephan Frings Prof. Nathaniel Heintz, MD, PhD, New York Prof. Dr. Joachim Kirsch Prof. Dr. Arthur Konnerth, München PD Dr. Georg Köhr Prof. Mark P. Mattson, MD, PhD, Baltimore Prof. Dr. Hannah Monyer Prof. Dr. Hans-Christian Pape, Magdeburg Prof. Dr. Christof Niehrs Prof. Dr. Mart Saarma, Helsinki Dr. Gerhard Schratt Prof. Stephen G. Waxman, MD, PhD, New Haven Prof. Dr. Christoph Schuster Prof. Dr. Markus Schwaninger Prof. Dr. Klaus Unsicker Prof. Dr. Joachim Wittbrodt

5 Research groups / IZN Investigators

IZN Investigators Department of Pharmacology

University of Heidelberg Rohini Kuner Molecular mechanisms underlying chronic pain Faculty of Medicine Heidelberg Stefan Offermanns Department of Anatomy and Cell Biology Cellular signaling Markus Schwaninger Uwe Ernsberger Gene regulation in cerebral ischemia Development of nerve cells and generation of neuronal diversity Department of Human Genetics Joachim Kirsch Molecular cell biology of synapses Gudrun Rappold Thomas Kuner Molecular pathogenesis of genetic disorders Structure and function of synapses Siegfried Mense Department of Neuropathology Neurobiological mechanisms of chronic muscle pain Andreas von Deimling Gerhard Schratt Molecular genetics of tumours of the central (CNS) and The role of microRNAs in synaptic development and the peripheral nervous system (PNS) plasticity Horst Simon Neurology Differentiation and survival of mesencephalic dopaminergic neurons André Rupp Auditory evoked fields, auditory modelling, and psy- Kerry Tucker choacoustics Early events in neurogenesis and nerve development in the central and peripheral nervous systems Wolfgang Wick Biology and experimental therapy of malignant glioma Klaus Unsicker Faculty of Medicine Mannheim TGF-ß, FGF and neurotrophins in neural development and functions Anaesthesiology

Department of Physiology and Pathophysiology Martin Schmelz Translational pain research Andreas Draguhn Functional analysis of cortical neuronal networks Faculty of Biosciences Ulrich Misgeld Plasticity of GABAA receptor mediated inhibition Centre for Biochemistry

Thomas Söllner Molecular mechanisms mediating regulated exocytosis

6 Research groups / IZN Investigators

Department of Neurobiology Thomas Holstein The origin of the metazoan nervous system and body Hilmar Bading plan Nuclear calcium signaling in the dialogue between Joachim Wittbrodt synapse and nucleus: Role in neuronal survival and Vertebrate retina development and differentiation memory Francesca Ciccolini Faculty of Behavioural and Cultural Studies Proliferation and differentiation of neural stem cells Institute for Psychology Christoph Schuster Mechanisms of experience-dependent synaptic plasti- Christian Fiebach city, learning and memory, and memory extinction Cognitive neuroscience: Neural mechanisms of higher cognitive functions Department of Clinical Neurobiology Sabina Pauen Early childhood brain and cognitive development Hannah Monyer Neuronal synchrony and plasticity Faculty of Mathematics and Computer Science Andrei Rozov Mechanisms of short lasting synaptic plasticity Institute for Computer Science

Gabriel Wittum Institute of Pharmacy and Molecular Biotechnology Computational neuroscience as part of the chair simulation in technology Ulrike Müller Functional genomics of neurodegenerative diseases¸ Alzheimer’s disease, physiological and pathological Central Institute for Mental Health Mannheim

function of APP family proteins Dusan Bartsch Centre for Molecular Biology Heidelberg Molecular and cellular basis of normal and pathologic cognition, learning and memory, and mental retardati- Stefan Kins on Alzheimer’s disease, APP function and transport Martin Bohus Pain perception, learning and memory, and treatment Department of Zoology development for patients with trauma-related disorders G. Elisabeth Pollerberg Herta Flor Growth and orientation of axons in the developing ner- Learning and neuronal plasticity vous system Peter Gass Stephan Frings Animal models of psychiatric disorders Signal transduction in sensory neurons Andreas Meyer-Lindenberg Translational neurogenetics of psychiatric disorders

7 Research groups / IZN Investigators

Rainer Spanagel European Molecular Biology Laboratory The neurobiology of drug abuse Detlev Arendt Max-Planck Institute for Medical Research The evolution of the animal central nervous system Darren Gilmour Winfried Denk The role of chemokine-mediated tissue migration in Structure of and activity in neuronal networks. generating mechanosensory organs in the zebrafish lateral line Thomas Euler Signal processing in the retina Georg Köhr Cross talk between excitatory and inhibitory synapses and ion channel and signalling function of NMDA receptors Peter Seeburg Fast excitatory neurotransmission, synaptic plasticity and learning Hartwig Spors Spatio-temporal patterns of neuronal activity in the healthy and diseased brain Rolf Sprengel The role of glutamate receptors in hippocampus mediated learning, emotional behaviors and mood disorders Veit Witzemann Molecular anatomy of the developing neuromuscular junction

German Cancer Research Center

Christof Niehrs Molecular embryology Günther Schütz Molecular genetics of signal-dependent gene expression Otmar D. Wiestler Molecular neuropathology of human brain tumors / surgical neuropathology of CNS tumors

8 Concept and Structure of the IZN

9 10 Introduction

Concept and Mission of the IZN

The Interdisciplinary Center for Neurosciences (IZN) was other research units both within and external to the Uni- founded in 2000 as a research network incorporating neu- versity of Heidelberg that collectively formed the IZN’s roscientists from all faculties and local research institutions ’ring‘ structure. in Heidelberg. Its mission is to enhance brain research, to At present, the IZN encompasses all research activities coordinate technology transfer and to improve graduate in the neurosciences in the Heidelberg/Mannheim area, and post-graduate education in the neurosciences. Origi- providing a forum for scientific exchange at all levels. Our nally consisting of three ’core’ institutes, including the weekly seminar series brings together junior and senior Departments of Neurobiology, Neuroanatomy, and Clini- scientists with diverse neuroscience backgrounds. Annual cal Neurobiology, the IZN has been joined by groups from retreats foster extensive communication and planning of the Departments of Physiology and Pathophysiology, common projects, and are complemented by regular, infor- Anatomy and Medical Cell Biology, as well as by various mal exchanges between group leaders and the IZN Board

Scienti c Advisory Board Acting Director Rectorate

Board of Directors 11 Senior and 2 Junior

IZN Administration

IZN Investigators

Faculty of Faculty of Faculty of Medical Medical Central Max-Planck German European Biosciences Mathematics Behavioral Faculty of Faculty of Institute for Institute Cancer Molecular and and Cultural Heidelberg Mannheim Mental for Medical Research Laboratory Computer Studies Health Research Center Sciences Mannheim 10 1 2 15 2 6 8 3 2

Excellence- BMBF DFG DFG DFG DFG DFG DFG HBIGS cluster Bernstein Klinische SFB 636 SFB 488 Graduate Forscher- Forscher- Cell Forscher- College 791 gruppe 577 gruppe 643 Networks gruppe 107

11 Introduction

Concept and Mission of the IZN Developmental Molecular Behavioural Computational Neuroscience and Physiology and Basic Neuroscience Neuroscience Genetics Imaging Sciences

Neurogenetic Cognitive Diseases Disorders Brain and Neuro- Tumors Addiction degenerative Diseases Applications AD, PD, ... Stroke Pain

Aging

Gene Pharma- Therapy cology and Regeneration Drug- and Repair Design Perspectives Behavior Pain Therapy Therapy Stem Cell Therapy

12 Introduction

of Directors. The IZN facilitates the common usage of spe- and methodological innovations have meant that the bor- cialized equipment and access to resources, and supports der between molecular and cellular neurosciences, on one joint funding schemes such as Sonderforschungsbereiche, hand, and behavioral/systems neurosciences, on the other, Forschergruppen, and Graduierten Kollegien. It shapes is becoming progressively more permeable. Secondly, the future of the local neuroscience community by partici- translational neurosciences are becoming more signifi- pating in search committees for new junior and senior sci- cant and powerful insofar as investigators are increasingly entists as well as by helping to decide upon new directions able to transfer knowledge from the basic neurosciences of research in the University of Heidelberg’s molecular life into clinical contexts. The IZN is actively involved in both sciences. The IZN runs a graduate program leading to the of these promising advancements. Prominent examples of degree ‘MSc Molecular Bioscience, Major Neuroscience‘, this involvement include the Collaborative Research Cen- and a PhD program. The IZN also plays a very active role ter ‘SFB 636‘ in which clinical and basic scientists are jointly in the university’s Cluster of Excellence ‘Cellular Networks: investigating the phenomena of learning and memory in From Molecular Mechanisms to a Quantitative Understan- the context of psychopathological processes. Additionally, ding of Complex Functions‘ (CellNetworks) and in the re- the SFB 636 has initiated a graduate school for translatio- search network for the ‘Quantitative Analysis of Molecular nal neurosciences, which will foster cooperative research and Cellular Biological Systems‘ (BIOQUANT). and interaction between clinical and basic neuroscientists. While Heidelberg has traditionally been regarded as a lea- In 2007, the Directors of the IZN have, together with the ding center in molecular and cellular biology, the IZN now IZN Scientific Advisory Board, reorganized the structure of includes several groups working on transgenic animal mo- the IZN to further improve its presence and impact in the dels, in vivo recording and imaging, and behavioral testing. international scientific community. Strategic planning, co- This systems-based approach has already been expanded ordinating future, high-profile faculty appointments, and to include new groups from the ZI in Mannheim and will establishing new funding initiatives in Heidelberg and be further improved by the creation of a new research Mannheim all require the active participation of a broad group for behavioral/systems neuroscience. and diverse group of neuroscientists extending beyond traditional institutional boundaries. To this end, a new In summary, the IZN has extended its scope of enquiry Board of Directors has been selected, representing most while simultaneously improving its capacity to conduct re- of the institutions and faculties that participate in the IZN. search in particular behavioral/systems and translational neurosciences. Its new structure is efficient as well as mo- These institutions are: the University of Heidelberg, the dern, and will assist both junior and senior investigators in University Hospital, the Max Planck Institute for Medical the Heidelberg/Mannheim area in addressing future chal- Research, the German Cancer Research Center (DKFZ), the lenges in the neurosciences. European Molecular Biology Laboratory (EMBL), and the Central Institute of Mental Health (ZI) in Mannheim. Ad- ditionally, the distinction between members of the IZN’s ‘core‘ and ‘ring‘ structures has been abolished in favor of a minimal hierarchy currently comprised of 51 ‘IZN Investi- gators‘ selected on the basis of their scientific merits. The IZN Investigators are introduced in the Research Profiles (pp 35).

Such organizational modifications acknowledge two recent developments in the neurosciences. First, conceptual

13 Appointments

Developments in the IZN since 2003: Darren Gilmour came to the European Molecular Biology Laboratory Appointments (EMBL) Heidelberg as a Group Leader in 2004.

During the period 2003 to 2008, the IZN has further conso- Hans-Hermann Gerdes lidated and gained international visibility. Scientists at the took a professorship at the University of Bergen, Nor- IZN have been very successful with major discoveries from way, Department of Biomedicine. the molecular to the systems level. Several IZN scientists have been appointed to positions outside Heidelberg, and Thomas Holstein a number of new recruitments to the Heidelberg neuro- was appointed to a professorship at the Department of science community could be made. Various members of Molecular Evolution and Genomics, Zoological Institute, the IZN have received honourable prizes and awards. University of Heidelberg in 2004.

Detlev Arendt Harald Hutter was appointed to a professorship in Zoology at the Uni- left the Max Planck Institute for Medical Research in versity of Heidelberg in 2008. 2005 to become an associate professor at the Simon Fraser University, Department of Biological Sciences, Martin Bohus Burnaby, Canada. was appointed to a chair in Psychosomatics and Psycho- therapy, University of Heidelberg, and Director of the Stefan Kins Dept. of Psychosomatics and Psychotherapy, Central became Group Leader at the ZMBH in 2007; in 2008 he Institute of Mental Health. was appointed to a chair for Human Biology and Hu- man Genetics at the TU Kaiserslautern. Andreas von Deimling became chair of the Department of Neuropathology, Georg Köhr Institute of Pathology, University of Heidelberg, in 2007. became Research Group Leader at the Max Planck Insti- tute for Medical Research in 2007. Thomas Euler became Research Group Leader at the Max Planck Insti- Rohini Kuner tute for Medical Research in 2006. was appointed to a professorship in Pharmacology & Toxicology at the University of Heidelberg in 2006. Dirk Feldmeyer took a professorship at the Federal Research Centre Thomas Kuner Jülich. in 2006 was appointed as full professor at the Institute for Anatomy and Cell Biology, University of Heidelberg. Rainer Friedrich MPI for Medical Research, in 2005 was offered a leading Andreas Meyer-Lindenberg position at the Miescher Institute Basel. was appointed to the chair in psychiatry at the Central Institute of Mental Health in Mannheim, as the succes- Peter Gass sor of Fritz Henn. became extraordinary professor of Psychiatry at the Med. Faculty Mannheim, consultant and research group Ulrike Müller leader at the Central Institute of Mental Health in 2004. became a professor for Functional Genomics at the In-

14 Appointments

stitute for Pharmacy and Molecular Biotechnology, Wolfgang Wick IPMB, at the University of Heidelberg in 2005. was appointed to a professorship at the Neurology Cli- nic of Heidelberg University Hospital, Department of Renato Paro Neurooncology, in 2007. left the ZMBH to the ETH Zürich to become founding director of the Center of Biosystems Science and Engi- Bill Wisden neering of the ETH Zürich, located in Basel. took a professorship in Neurobiology at the University of Aberdeen, Scotland. Gudrun Rappold was appointed to a professorship and director of the Gabriel Wittum newly established Department of Human Molecular Was appointed to a chair for Modelling and Simulation Genetics, Institute of Human Genetics, University of Hei- at the Institute for Informatics, Johann Wolfgang Goe- delberg. the-University, Frankfurt/Main, in 2008.

Andrei Rozov Jochen Wittbrodt left Heidelberg to become a senior lecturer at the Cen- Accepted offers from the Universities of Karlsruhe as di- tre for Neuroscience, University of Dundee, beginning rector at the Research Center Karlsruhe, and Heidelberg of 2008. as professor of Developmental Biology at the Heidel- berg Institute of Zoology (HIZ). Gerhard Schratt and Kerry Tucker were appointed as DFG funded Junior Group Leaders in the Sonderforschungsbereich 488, University of Heidel- berg, and established their groups. We welcome all new group leaders and wish the leaving colleagues all the best with their new and challenging Christoph Schuster positions. received a professorship of the Hertie Foundation within the Excellence Program in Neurosciences. He took a professorship in Developmental Neurobiology at the University of Heidelberg, Faculty of Biosciences, and established his group in Neurobiology.

Markus Schwaninger moved from the Neurology Clinic of Heidelberg Univer- sity Hospital to a professorship in Pharmacology at the Faculty of Biology and Pharmacy.

Thomas Söllner was appointed as full professor at the Biochemistry Cen- ter, University of Heidelberg, in 2005.

Hartwig Spors moved to the Max Planck Institute of Biophysics in Frankfurt/M. in 2008 as a Research Group Leader.

15 Awards

Awards Member of the German Academy of Sciences Leopoldina (2008) Hilmar Bading Stephan Frings ERC Advanced Grant (2008) Lehrpreis des Landes Baden-Württemberg (2007) Martin Bohus Thomas Holstein Established Investigator Award (2003) Borderline Personality Disorder Research Foundation, Member of the Heidelberg Academy of Sciences and New York Humanities (2007) Research Award (2003) Rohini Kuner International Society for the Investigation and Rudolf-Buchheim Annual Prize (2005) Teaching of Dialectical Behavioral Therapy, Boston German Society for Experimental and Clinical Psychotherapy Award (2004) Pharmacology and Toxicology German Association of Psychiatry, Psychotherapy Chica & Heinz Schaller Prize (2006) and Neurology Research Award (2006) Outstanding Research Award (2005) Rotary Club, Heidelberg International Society for the Investigation and Teaching of Dialectical Behavioural Therapy, First Prize for Basic Research on Pain (2007) Washington German Pain Society (DGSS) Ingrid zu Solms Wissenschaftspreis (2008) Winfried Denk Gottfried Wilhelm Leibniz Prize (2003) Siegfried Mense German Research Council Muscle Pain Award (2004) Alden Spencer Award (2006) Wiss. Arbeitskreis Muskel und Schmerz Columbia University, New York Honarary Pain Award (2006) Kavli Lecture (2006) Deutsche Gesellschaft für Schmerztherapie Society for Neuroscience Andreas Meyer-Lindenberg Henri Sack Lecture (2008) Bench-to-Bedside Award (2004-2006) Cornell University, Ithaca NIMH/ORD/NIAAA Herta Flor Roche/Nature Medicine Award for Translational Research Award (2003) Neuroscience (2006) German Society for Neurotraumatology and Clinical Joel Elkes International Award for Clinical Research Neuropsychology (2006) Award for Basic Research (2004) American College of Neuropsychopharmacology State Baden-Württemberg E. Bennett Research Award (2007) ERC Advanced Grant (2008) Society of Biological Psychiatry Marsilius College Fellow (2008) University of Heidelberg

16 Awards

Hannah Monyer Christoph Schuster Gottfried Wilhelm Leibniz Prize (2004) Neurosciences Excellence Award (2004) German Research Council Hertie-Foundation Prix Franco-Allemand Gay-Lussac–Humboldt (2005) Peter Seeburg Member of the German Academy of Sciences InBev-Baillet Latour Health Prize (2007) Leopoldina (2006) Rainer Spanagel Philip Morris Foundation Research Award (2006) Sir Hans Krebs Award (2003) Member of the Heidelberg Academy of Sciences and Humanities (2008) Albrecht-Ludwig-Berblinger Award (2005) James B. Isaacson Award (2008) Ulrike Müller Alzheimer Forschungspreis (2008) Hartwig Spors Hans und Ilse Breuer Stiftung Otto-Hahn-Medal (2003) Max-Planck Society Christof Niehrs Gottfried Wilhelm Leibniz Prize (2003) Klaus Unsicker German Research Council Honorary Doctorate Univ. Marburg (2003) Member of the Heidelberg Academy of Sciences and Honorary Member of the Romanian Society for Cell Humanities (2007) Biology (2007) Gudrun Rappold Wolfgang Wick European Society of Human Genetics Award (2003) Novartis-Award for Clinical Research (2003) (S. Schiller) Sibylle-Assmus Award for Neurooncology (2005) Young Scientist Award (2004) Pette Award (2006) European Society of Human Genetics (N. Sabherwal) German Society of Neurology Martin Schmelz Young Investigator Award (2008) Sertürner Award (2003) American Society of Clinical Oncology (M. Weiler) Award of the German Society for Anesthesiology Gabriel Wittum (2003) doIT Software Award (2005) Collaborative Research Award (2003) International Association for the Study of Pain Gerhard Schratt Career development Award (CDA) (2006) Human Frontier Science Program Analytica Forschungspreis (2008) (jointly with Albert Sickmann)

17 IZN Funding and grant giving bodies

IZN Funding and grant giving bodies

Basic funding Basic funding for IZN research groups is provided by their home institutions: • University of Heidelberg • University Clinics Heidelberg • University Clinics Mannheim • Central Institute for Mental Health, Mannheim (ZI) • German Cancer Research Center (DKFZ) • Max-Planck Institute for Medical Research (MPIMF) • European Molecular Biology Laboratoy (EMBL)

Grant giving Organizations Additional funding (‘Drittmittel‘) is provided by the fol- • Génoscope, France lowing grant giving organizations: • German-Israeli Foundation (GIF) • Hans und Ilse Breuer Stiftung • Abbott GmbH • Heidelberg Academy of Sciences • Alexander von Humboldt-Stiftung • Hertie Stiftung • ALS Association, USA • Human Frontier Science Program (HFSP) • Alzheimer Forschung Initiative e.V. • Klaus Tschira Stiftung • American Tinnitus Association, USA • Landesgraduiertenförderung Baden-Württemberg • Association for International Cancer Research (AICR), UK • Landesstiftung Baden Württemberg • BASF-Foundation • Manfred Lautenschläger Stiftung • Becton Dickinson GmbH • Merz AG • Boehringer Ingelheim Fonds • Michael J. Fox Foundation, USA • BPDRF, USA • Ministerium für Wissenschaft, Forschung und Kunst • Bundesministerium für Bildung und Forschung (BMBF) Baden-Württemberg • Cluster of Excellence, Cellular Networks, University of • National Institute of Drug Abuse (NIDA), USA Heidelberg • National Institutes of Health (NIH), USA • Deutsche Forschungsgemeinschaft (DFG) • National Parkinson Foundation, USA • Deutsche José Carreras Leukämie-Stiftung eV • Netzwerk Alternsforschung (NAR) • Deutsche Krebshilfe e.V. • Pfizer Pharma GmbH • Deutscher Akademischer Austauschdienst (DAAD) • PROSKELIA S.A.S., France • Dietmar-Hopp-Stiftung • Schering GmbH • Eli Lilly Foundation, USA • Schilling Stiftung • European Molecular Biology Organisation (EMBO) • Storz, Swizerland • European Society for Pediatric Endocrinology (ESPE) • Tinnitus Research Initiative • European Union (EU) • Volkswagenstiftung • Forschungsschwerpunktprogramm des Landes Baden- • Wilhelm Sander Stiftung Württemberg • Fritz Thyssen Stiftung

18 Guest scientists

Guest scientists Kirsch Many colleagues from abroad have taken advantage to Dean Smith (2008), Texas Tech University, Lubbock, work at the laboratories of various IZN groups for a certain USA period of time, to discuss ideas and exchange methods, Mense and to make use of the common infrastructure provided by the IZN. D.B. Simons (2001-2003), Emory University, Atlanta, USA Bading Heinz Steffens (2003-2004), Physiology, Universität Peter Vanhoutte (2003), MRC Lab. of Molecular Biology, Göttingen, Germany Cambridge, UK Siavash Gholami (2004-2005), Shiraz University, Iran Marvin Steijaert (2004), Technische Universiteit, Marucia Chacur (2007), Sao Paulo University, Brazil Eindhoven, The Netherlands Monyer Dmitri Tkachev (2004), Babraham Institute, Cambridge, Smaragda Lambrianou (2003), Pasteur Institute, Paris, UK France Xiaoxuan Qi (2007), University of Saint Andrews, UK Roberto Bruzzone (2002-2004), Pasteur Institute, Paris, Draguhn France Rafael Gutierrez (2003), Fisiologia, Biofisca y Neuro- Mark Cunningham (2004-2006) University of Leeds, UK sciencias, Centro de Investigacion y Estudios Ruth Benavides (2004), Instiuto Cajal, Madris, Spain Avanzados del I.P.N., Mexico Pablo Pelegrin (2006), University of Sheffield, UK Frings Jason McLean (2006), Columbia University, New York, Joseph Lynch (2003), University of Queensland, USA Australia Brendan Watson (2006), Columbia University, New Gilmour York, USA Michael Granato (2006-2007), University of Matthew McGinley (2007), Portland, Oregon, USA Pennsylvania, USA Roger Traub (2007-2008), SUNY Downstate Medical Holstein Center, Brooklyn, N.Y., USA Mihaela Žigman (2006-2007), Institute of Molecular Pierre Pratley (2008), University of Groningen, The Biotechnology of the Austrian Academy of Science, Netherlands Vienna Niehrs Toshika Fujisawa (2007-2008), National Institute of Wei Wu (2008), Tshingua University, China Genetics, Mishima, Japan Pauen David Vactor (2008), Harvard Medical School, Boston, USA Laraine McDonough, Brooklyn College, New York, USA Shan Wang (2008), National Institute of Genetics, Pollerberg Mishima, Japan Galina Schevzov (2006), University of Sidney, Australia Susanne Theiss (2006), University of Vancouver, Canada

19 Guest scientists

Francisco Ropero-Padilla (2007), University of Madrid, Dorota Dudys (2004), Dept. Of Pharmacology, Institute Spain of Phamacology, Krakau, Poland Shinji Fushiki (2008), University of Kyoto, Japan Schu-Fee Yang (2004), MCB Program, DKFZ, University of Heidelberg, DKFZ, Heidelberg, Germany Rappold Nato Kotaria (2004-2005), Group of Chemical Neuro- Zilin Zhong (2007-2008), Anhui Medical University, anatomy, Institute of Physiology, Tbilisi, Georgia Hefei, Anhui, PR China Maxim Sheroziya (2004-2005, 2007), Institute of Schmelz Higher Nervous Activity and Neurophysioloy, Martin Angst (2003), Stanford University, USA Russian Academy of Science, Moscow, Russia Schratt Shlomi Krispin (2007), Hebrew University, Jerusalem, Israel Mette Christensen (2007), University of Kopenhagen, Denmark Von Deimling Schuster Prabal Deb (2007), All India Institute of Medical Sciences, New Delhi, India Hiroshi Kuromi (2005), Gunma University, Maebashi, Japan Wittbrodt Yoshiaki Kidokoro (2006-2007), University of California Pilar Esteve (2004), Instituto Cajal, Madrid, Spain at Los Angeles, USA Juan-Ramon Martinez-Morales (2008), University of Schütz Sevilla, Spain Ute Moll (2006-2007), Stonybrook University, New York, Wittum USA Randolph E. Bank, UCSD, La Jolla, USA Simon J. Xu, PennState, College Park, USA Richard Dyck (2004-2005), University of Calgary, Roger Traub, SUNY, New York, USA Canada Mary Wheeler, ICES, UTexas, Austin, USA Spanagel Dai Stephens, University of Sussex, UK Theodora Duka, University of Sussex, UK Tucker Almut M. Ellwanger (2005-2006), Mt. Holyoke College, USA Fabiola Zelada Gonzalez (2005), Zoology, Universität Heidelberg. Germany Unsicker Noga Ratner (2003), Katzir Highschool, Rehovot, Israel Tuna Cakar (2003), Biological and Bioengeneering Program, Sabanci University, Turkey

20 Collaborations

Collaborations Universität Heidelberg, Germany Learning in Flies IZN Investigators have build up and continuously expand Rainer Spanagel, ZI Mannheim, Germany a widespread network of collaborations, both internal and Addiction external, in which they intensely interact with groups from all fields of neurosciences. Andre Fiala, Universität Würzburg, Germany Learning in Flies Arendt Mathias Bähr, Institut für Neurobiologie, Universität Franςois Nedelec/ Ernst Stelzer, EMBL, Germany Göttingen, Germany Mathematical modelling of Platynereis swimming Neuronal Survival Harald zur Hausen, Freie Universität Berlin, Germany Amin Rustom, AG Spatz, MPI für Metallforschung Electron optic characterisation of Platynereis photo- Stuttgart, Universität Heidelberg, Germany receptor cells Role of TNTs in Ca-Signalling Günter Purschke, Universität Osnabrück, Germany William Wisden, University of Aberdeen, Institute of Electron optic characterisation of Platynereis photo- Medical Sciences, UK receptor cells De novo methyltransferase Nicole Rebscher, Universität Marburg, Germany Patrick Descombes, Genomics Platform at the Platynereis germ cells University of Geneva, Switzerland Mechanisms of neuroprotection Maria Ina Arnone, Stazione Zoologica, Naples, Italy Photoreceptor cell evolution Malte Wittmann, Karolinska Institute, Stockholm, Sweden Michael Akam, Cambridge University, UK Dynamics of nuclear geometry Patterning of the bilaterian body plan Lars Christian Rønn, NeuroSearch, Ballerup, Denmark Gregor Bucher, Universität Bayreuth, Germany Gene regulation in ischemia Patterning of the bilaterian body plan Armando Genazzani, Dept. of Chemical, Food, Rolf Urbach, Universität Mainz, Germany Pharmaceutical and Pharmacological Sciences Patterning of the bilaterian body plan (DiSCAFF) at the Università del Piemonte Orientale A. Avogadro, Novara, Italy Bading Neuroprotective mechanisms in the cerebellum Christof Niehrs, Heidelberg, Germany Functional characterization of a novel Wnt regulator Bartsch in the central nervous system Eric Kandel, Columbia University, USA Klaus Unsicker, Institut für Neuroanatomie und Minibrain Zellbiologie, Universität Heidelberg, Germany Jörg Striessnig, Universität Innsbruck, Österreich Development of neuroendocrine derivatives of the Cav1.3 neural crest (NC) Michael Kiebler, Universität Wien, Österreich Günther Schütz, DKFZ, Heidelberg, Germany Neuronal differentiation The role of the nuclear receptor ´tailless´ in development and adult neural stem cells Cornelius Gross, EMBL, Monterotondo, Italy Stress and serotonin Christoph Schuster, Institut für Neurobiologie,

21 Collaborations

Marlies Knipper, Universität Tübingen, Germany Magnus Institute of Neuroscience, University Cav1.3 and hearing Medical Center, Utrecht, The Netherlands Matteo E. Mangoni, University of Montpellier, France Pain perception in Posttraumatic Stress Disorders Cav1.3 in heart Andrea Lüthi/ I. Mansui, ETH Zürich, Friedrich Miescher Susumu Tonegawa, Massachusetts Institute of Institut, Basel, Schweiz Technology, Boston, USA Entwicklung von Tiermodellen zur Stressregulation Conditional mutations in rat bei Borderline-Störungen (BPDRF) Roland Buelow, Open Monoclonal Technology (OMT) Ciccolini Inc., USA Zn finger nucleases Klaus Unsicker, Institut für Neuroanatomie und Zellbiologie, Universität Heidelberg, Germany Bohus Role of GDF 15 in neural stem cells Marsha M. Linehan, University of Washington, USA Günther Schütz, DKFZ, Heidelberg, Germany Treatment development Role of CREB and CREM in neural stem cells Thomas Lynch, Duke University, Durham, USA Kerry Tucker, Institut für Neuroanatomie und Conditioning in addictive behaviour Zellbiologie, Universität Heidelberg, Germany FACS sorting of neurons and stem cells from the em- Larry Siever, Mount Sinai School of Medicine, Dept. of bryonic mouse brain Psychiatry, Bronx VA Medical Centre, USA Genetic Consortium in BPD Anthony D. Ho, Innere Klinik V, Universität Heidelberg, Germany Mary C. Zanarini, McLean Hospital, Harvard Medical FACS sorting of neural precursors School, USA Genetics in borderline personality disorder A. Paula Monaghan, School of Medicine, University of Pittsburgh, USA Shelly McMain, Dept. of Psychiatry, University of Role of Tlx in neural stem cell maintenance Toronto, Canada Genetic Consortium in BPD Draguhn Ruth Lanius, University of Western Ontario, Canada Uwe Heinemann, Charité, Berlin, Germany fMRI of dissociative states Modulation of hippcoampal networks James D. Bremner, Dept. of Psychiatry, University of Klaus Willecke, Universität Bonn, Germany Atlanta, Georgia, USA Gap junctions and neuronal synchrony Neuroimaging studies of dysfunctional stress regulation in patients with traumatic experiences William Wisden, University of Aberdeen, Scotland, UK Network functions of inhibitory interneurons Bernet Elzinga, University of Amsterdam, The Netherlands Roger D. Traub, SUNY Health Center, New York, USA Pain perception in stress related disorders Modelling of network oscillations John Livesley, Dept. of Psychiatry, Faculty of Medicine, Ernsberger University of British Columbia, Vancouver, Canada Genetics of categorical and dimensional factors of Herrmann Rohrer, MPI für Hirnforschung, Frankfurt, personality disorders Germany Neuron subclass specification Herman G.M. Westenberg, Dept. of Psychiatry, Rudolf

22 Collaborations

Katrin Huber, Institut für Neuroanatomie und Angela Friederici, MPI für Kognitions- und Zellbiologie, Universität Heidelberg, Germany Neurowissenschaften, Leipzig, Germany Transcription factors in neuronal differentiation Neural mechanisms of language processing - con- Klaus Unsicker, Institut für Neuroanatomie und ceptual combination in sentence processing and Zellbiologie, Universität Heidelberg, Germany working memory Adrenal chromaffin cell differentiation Martin Reuter, Psychologisches Institut, Universität Hidemi Misawa, Tokyo Metropolitan Institute for Bonn, Germany Neuroscience, Tokyo, Japan Cognitive effects of neurotransmitter gene polymor- Regulation of cholinergic gene expression phisms Edward Smith, Dept. of Psychology, Columbia Euler University New York, USA Peter B. Detwiler, University of Washington, USA Conceptual combination in sentence processing and Diverse projects concerning retinal information pro- working memory cessing Hauke Heekeren, MPI für Bildungsforschung, Berlin, Silke Haverkamp, Institut für Neuroanatomie, MPI für Germany Hirnforschung, Frankfurt/M., Germany An fMRI study of cost-benefit analysis in the brain Color processing in the non-primate retina Flor Brendan J. O’Brian, University of Auckland , New Zealand Sodium channels in starburst amacrine cells Klaus van Ackern, Institut für Anästhesiologie, Universität Heidelberg, Germany Rowland W. Taylor, Oregon Health & Science KFO 107 University, Portland, WA, USA Direction-selective ganglion cells Martin Schmelz, Institut für Anästhesiologie, Universität Heidelberg, Germany Shigang He, Chinese Academy of Sciences, Beijing, KFO 107 China Receptive field properties of starburst amacrine cells Marcus Schiltenwolf, Institut für Orthopädie, in the rabbit retina Universität Heidelberg, Germany KFO 107 Jost B. Jonas/ Frank Schlichtenbrede, Universitäts- Augenklinik , Mannheim, Germany Eike Martin, Institut für Anästhesiologie, Toxicity Assessment of intravitreal Triamcinolone and Universität Heidelberg, Germany Bevacizumab in an ex-vivo mouse model KFO 107 Thomas Kuner, Institut für Anatomie und Zellbiologie, Gisela Skopp, Institut für Rechtsmedizin, Universität Universität Heidelberg, Germany Heidelberg, Germany Chloride gradients in retinal neurons KFO 107 Hubert J. Bardenheuer, Institut für Anästhesiologie, Fiebach Universität Heidelberg, Germany Mark D’Esposito, Helen Wills Neuroscience Institute KFO 107 and Dept. of Psychology, University of California, Andreas Heinz, Charité – Universitätsmedizin Berlin, Berkeley, USA Germany Neurocognitive bases of verbal working memory IMAGEN

23 Collaborations

Christian Buechel, Universitätsklinikum Hamburg- Jörg Trojanl, Otto-Selz-Institut, Universität Eppendorf, Germany Mannheim, Germany IMAGEN SOMAPS Gunther Schumann, Institute of Psychiatry, King’s Lars Arendt-Nielsen, Dept. of Health Science and College, London, UK Technology, University of Aalborg, Denmark KFO 107, IMAGEN SOMAPS Trevor W. Robbins, University of Cambridge, UK Laura Petrini, Dept. of Health Science and Technology, IMAGEN University of Aalborg, Denmark David N. Stephens, University of Sussex, UK SOMAPS IMAGEN Jan R. Buitenweg, Institute for Biomedical Technology Tomas Paus, University of Nottingham, UK (BMTI), University of Twente, The Netherlands IMAGEN SOMAPS John Rogers, Delosis Ltd.(SME), UK Peter H. Veltink, Institute for Biomedical Technology IMAGEN (BMTI), University of Twente, The Netherlands Romain Valabrègue, SCITO S.A. (SME), France SOMAPS IMAGEN Soledad Ballesteros, UNED Research Institute (CEEN), Jean Baptiste Poline, Commissariat à l’Energie, Dept. of Basic Psychology II, Universidad Nacional Atomique, France de Educación a Distancia (UNED), Spain IMAGEN SOMAPS Mark Lathrop, Consortium National de Recherche en Jose Manuel Reales, UNED Research Institute (CEEN), Genomique, France Dept. of Basic Psychology II, Universidad Nacional IMAGEN de Educación a Distancia (UNED), Spain SOMAPS Patrick Konstant, PERTIMM PERTINENT ET IMMEDIAT (SME), France Beatriz García, UNED Research Institute (CEEN), Dept. IMAGEN of Basic Psychology II, Universidad Nacional de Educación a Distancia (UNED), Spain Jean-Luc Martinot, INSERM - CEA ERM, Orsay, France SOMAPS IMAGEN Tormod Thomsen, NordicNeuroLab (SME), Norway Frings IMAGEN Thomas Kuner/ Hartwig Spors/ Hannah Monyer, Kathinka Evers, Uppsala University, Sweden Institut für Anatomie und Zellbiologie, Universität IMAGEN Heidelberg/ MPI für Medizinische Forschung, Hugh Garavan, Trinity College Dublin, Irland Heidelberg/ Institut für Neurobiologie, Universität IMAGEN Heidelberg, Germany Rupert Hoelzl, Otto-Selz-Institut, Universität DFG Research group “Information processing in the Mannheim, Germany olfactory system” SOMAPS Martina Schnölzer, DKFZ, Heidelberg, Germany Dieter Kleinböhll, Otto-Selz-Institut, Universität Proteomic analysis of olfactory neurons Mannheim, Germany Joseph Lynch, University of Queensland, Australia SOMAPS Chloride imaging methods

24 Collaborations

Jon Bradley, Université Paris 5, France Kins Chloride channel cloning Joachim Kirsch, Institut für Anatomie und Zellbiologie, Johannes Reisert, Monell Taste and Smell Center, USA Universität Heidelberg, Germany Chloride channel biophysics Subcellular localization of APP/APLPs Thomas Gensch, Jülich Research Center, Germany Chloride homeostasis in sensory neurons Kirsch Ulrike Müller, Institut für Pharmazie und Molekulare Gass Biotechnologie, Universität Heidelberg, Germany Marco Riva, Dept. of Pharmacology Science, University Genetic manipulation of mice of Milano, Italy Jochen Herms, Ludwig-Maximilians-Universität, Neuronal signalling and Depression München, Germany Rainer Hellweg, Psychiatrische Klinik, Charité, Berlin, APP mediated synaptogenesis Germany Marino Zerial, MPI für Molekulare Zellbiologie und Neurotrophins and Depression Genetik, Dresden, Germany Susana Aznar, Dept. of Neurobiology, University of Rab5 dependent Endocytosis Copenhagen, Denmark Ruppert Egensperger, Ludwig-Maximilians-Universtität, Serotonin and Depression München, Germany Electronmicroscopic anlyses of APP transport vesic- Gilmour les Jochen Wittbrodt, EMBL, Heidelberg, Germany Scott Brady/ Gerardo Morfini, University of Illinois, Three-Dimensional imaging/tracking of Cell Move- Chicago, USA ments in embryos using DSLM Molecular interaction of APP and Kinesin Sangram Sisodia, University of Chicago, Chicago, USA Holstein Molecular interaction of APP and Kinesin Brad Amos, Cancer research UK, imaging and Ole Kjaerulff, Copenhagen University, Copenhagen, microscopy laboratory, University of Cambridge , UK Denmark EURESCO Multi-harmonic Generation imaging using Electrophysiological studies of APP transgenic Dro- a confocal microscope sophila larvae Hans C. Gerritsen, Dept. of Molecular Biophysics, Dept. Gunther Merdes, Institute for System Biology, Basel, of Physics & Astronomy, Science Faculty, Universiteit Switzerland van Utrecht, The Netherlands Generation of transgenic flies Maarten Balzar, Nikon Instruments Europe B.V., Takuya Sasaki, University of Tokushima, Tokushima, Product Support Group, The Netherlands Japan Hans Bode, Dept. of Developmental Biology, Genetic manipulation of Rab3GAP University of Irvine, USA Eckart Friauf, Universität Kaiserslautern, Germany Hydra Genome Project Organotypic cultures of the brain stem Dan Rokhsar, Dept. of Molecular and Cell Biology, Heinrich Betz, MPI für Hirnforschung, Frankfurt, University of Berkeley, USA Germany Hydra Genome Project Cellular analysis of collybistin knock-out mice

25 Collaborations

Dean Smith, Dept. of Physiology, Texas Tech University, Shanelle W. Ko/ Min Zhuo, Dept. of Physiology, Faculty USA of Medicine, University of Toronto, Canada Electrical properties of newborn and immature neu- Forebrain plasticity in pain rons Anthony Dickenson, Dept. of Neuropharmacology, Peter Lauf, Cell Biophysics Group, Dept. of Pathology, University College, London, UK Wright State University, Dayton, USA Tumor-induced pain Regulation KCC2 in newborn and immature neurons Seifollah Ahmadi/ Hanns U. Zeilhofer, Institut für Pharmakologie und Toxikologie, Universitätsklinikum Köhr Erlangen, Germany Egidio D’Angelo, Dept. of Cellular-Molecular Physiology Synaptic plasticity in pain and Pharmacology, University of Pavia, Italy Gary Lewin, Molecular Physiology of Somatic Cerebellar plasticity Sensation, Max-Delbrück-Zentrum für Molekulare Chris I. De Zeeuw, Dept. of Neuroscience, University of Medizin, Berlin, Germany Rotterdam, The Netherlands Inflammation and pain Eyeblink conditioning Giovanni Marsciano/ Beat Lutz, Institut für Anita Lüthi, University of Lausanne, Switzerland Physiologische Chemie, Johannes Gutenberg- Long-term depression Universität, Mainz, Germany Øivind Hvalby/ Vidar Jensen, Dept. of Neurophysiology, Cannabinoid receptors and pain University of Oslo, Norway Robert Feil, Interfakultäres Institut für Biochemie, Dopamine-NMDA-Interactions Universitätsklinikum Tübingen, Tübingen, Germany cGK1-mechanisms in pain Kuner R Kuner T Bernhard Bettler/ E. Casanova, Dept. of Biomedicine, Institute of Physiology, University of Basel, George Augustine, Duke University Medical Center, Switzerland USA Endogenous control of pain by GABA-B1 Chloride signalling Silvia Arber, Biozentrum, University of Basel, Daniel Gitler, Ben Gurion University, Israel Switzerland Synaptic vesicle cycle Genetic labelling of sensory neurons Eckart Gundelfinger, Institut für Neurobiologie, Mike Costigan/ Clifford Woolf/ Irmgard Tegeder, Universität Leibzig, Germany Dept. of Anesthesia and Critical Care, Massachusetts CAZ proteins bassoon and piccolo in calyx General Hospital and Harvard Medical School, USA Benedikt Grothe, Ludwig-Maximilians-Universität, (Costigan/ Woolf)/ Goethe- Universität, Frankfurt, München, Germany Germany Auditory system Neuropathic pain in transgenic mice Rohini Kuner, Institut für Pharmakologie, Universität Sugiura Takeshi/ Gerald Gebhart, Dept. of Anesthesio- Heidelberg, Germany logy, Pittsburgh Center for Pain Research, University Virus-mediated perturbations of Pittsburgh, USA Peter Seeburg, MPI für Medizinische Forschung, Visceral pain in transgenic mice Heidelberg, Germany Glutamate receptor mouse models

26 Collaborations

Mense Bill Wisden, Institute of Medical Sciences, University of Thomas Unger, Center for Cardiovascular Research/ Aberdeen, UK Institut für Pharmakologie and Toxikologie, Charité, Mice with altered GABA receptors in parvalbumin- Berlin, Germany positive cells, AMPA receptor KO in cerebellar granu- Neuroinflammation and chronic muscle pain: role of le cells glial cells in central sensitisation Hans van Hooft, Institute of Neurobiology, University Lars Arendt-Nielsen, Center for Sensory-Motor of Amsterdam, The Netherlands Interaction, Aalborg, Denmark Analysis of 5HT3 EGFP transgenic mice NGF-induced hyperalgesia in muscle Roberto Bruzzone, Pasteur Institute, Paris, France Kazue Kumazawa, Dept. of Neuroscience II, Division Functional characterization of recombinant panne- of Stress Recognition and Response Research xins Institute of Environmental Medicine, Nagoya Reto Weiler, Institut für Biologie und Umweltwissen University, Japan schaften, Universität Oldenburg, Germany Glia cells and muscle pain Analysis of connexin and pannexin expression in Karl Messlinger, Institut für Physiologie und identified neurons in the retina Pathophysiologie, Peter Seeburg, MPI für Medizinische Forschung, Universität Erlangen-Nürnberg, Germany Heidelberg, Germany Environmental influences on muscle pain Conditional NR2B knockout mice. Generation of Bob Gerwin, Pain & Rehabilitation Medicine, transgenic mice with altered C-termini Bethesda, USA Andrei Rozov, MPI für Medizinische Forschung, Myofascial trigger points Heidelberg, Germany Functional characterization of calretinin-positive in- Monyer terneurons. Analysis of GluRD knockout mice Roger Traub, SUNY Health Center, New York, USA Markus Schwaninger, Institut für Pharmakologie, The role of AMPA receptors in GABAergic interneu- Universität Heidelberg, Germany rons for network synchronicity Stroke model in genetically altered mice Gary Westbrook, Vollum Institute, Portland, USA Study of defined cell types in the olfactory bulb; Müller exchange of Postdocs Heinrich Betz, MPI für Hirnforschung, Frankfurt, Gyorgy Buzsaki, Rutgers University, Newark, USA Germany Analysis of data obtained from in vivo recordings Glycine receptors Miles Whittington, University of Newcastle, UK Konrad Beyreuther, Zentrum für Molekulare Biologie The role of AMPA receptors in GABAergic interneu- Heidelberg, Universität Heidelberg , Germany rons for network synchronicity APP transport Nick Rawlins, Dept. Exp. Psychology, University of Sangram Sisodia, University of Chicago, USA Oxford, UK APP transgenic mice Behavioral studies of genetically altered mice Martin Korte, Technische Universität Braunschweig, Germany APP and synaptic function

27 Collaborations

Ulrich Zeilhofer, University of Zürich, Switzerland Oliver Müller, MPI für Molekulare Physiologie, Pharmacology of pain Dortmund, Germany David Wolfer, University of Zürich, Switzerland Microtubule-associated protein APC: impact on Mouse behavioural phenotyping growth cones Jochen Herms, Institut für Neuropathologie, Ludwig- Harald Hutter, MPI für Medizinische Forschung Maximilians-Universität, München, Germany Heidelberg, Germany Neuropathology of transgenic mice IgSF cell adhesion molecules: Role for axonal navigation in vertebrates and invertebrates Bart DeStrooper, University of Leuven, Belgium APP physiology Osborne Almeida/ I. Sotiropulos, MPI für Psychiatrie, München, Germany Benedikt Brors, DKFZ Heidelberg, Germany Cytoskeletal protein phosphorylation in Alzheimer Bioinformatics, transcriptome analysis disease Niehrs Joachim Spatz, MPI für Metallforschung, Stuttgart, Germany Thomas Holstein, Zoologisches Institut, Universität Nano-patterned proteins: impact on neurons Heidelberg, Germany Wnt signalling in Hydra, role of Hydra dkk Dieter Langosch, Technische Universität, München- Weihenstephan, Germany Pauen Properties of transmembrane domains of axonal proteins Usha Goswami, Cambridge University, UK Development of intermodal matching Jacob Piehler, Universität Frankfurt, Germany Nanopatterns Elisabetha Visalberghi, Rome Primate Center, Italy Tool use in infants and capucino monkeys Anthony Ho/ Sandra Gottschling, Innere Medizin V, Universität Heidelberg, Germany Josef Call, MPI Kognitions- und Neurowissenschaften, Adult human stem cell differentiation Leipzig, Germany Causal reasoning in infants and primates Gudrun Rappold, Institut für Humangenetik, Universität Heidelberg, Germany Stella Vosniadou, University of Athens, Greece MEGAP in growth cones Using analogies in educational context Konrad Beyreuther/ Tobias Hartmann, ZMBH, Pollerberg Heidelberg, Germany Stem cell differentiation in the retinal context Friedrich Propst, Institut für Biochemie und molekulare Zellbiologie, Universität Wien, Austria Benny Geiger, Weizmann Institute, Rehovot, Israel Analysis of the visual system of MAP1B deficient Intracellular signalling and micro-patterned substra- mice tes Klein, T., Keyence, Germany Enrique de la Rosa, Centro des Investigaciones in vivo imaging Biologicas, Spain Analysis of retinal cell lines with respect to apoptosis Fritz G. Rathjen, Max-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany Peter Sonderegger, University of Zürich, Switzerland Characterisation of the visual system of NrCAM defi- Interaction of CAMs cient mice

28 Collaborations

W. James Nelson, Stanford University, Beckmann Frank B. Gertler, Massachusetts Institute of Technology, Center, USA Boston, USA Impact of MAPs on axons Lamellipodin Susan McConnell, Stanford University, Biological Sciences, USA Rupp Embryonic development of the central nervous sys- Roy Patterson, Centre for Neural Basis of Hearing, tem Physiology Dept., Cambridge University, Great Britain George J. Weiner, University of Iowa, Iowa City, USA Pitch and Size Perception: Auditory Image Model Analysis of the visual system in DM-GRASP deficient and Magnetoencephalography mice Israel Nelken, Dept. of Neurobiology, The Alexander Patricia F. Maness, University of North Carolina, Silberman Institute of Life Sciences, The Hebrew Chapel Hill, USA University of Jerusalem, Israel CAM signalling and trafficking Comodulation Masking Release. Systematic comparison of intracellular Cat AI data and Neuromagnetic Claus Kremoser, MPI für Biologische Kybernetik, Recordings in Human Listeners Tübingen, Germany Genes involved in differentiation of retinal cells Georg Klump, Institut für Biologie, Carl von Ossietzky Universität Oldenburg, Germany Henning Bauch, Alfred-Wegener-Institut für Polar-und Comodulation Detection Difference: From the audi- Meeresforschung, Bremerhaven, Germany tory nerve to the auditory cortex. Auditory Models 3D and 4D imaging of axons in histotypic environ- and Neurophysiological Representation ment Stefan Uppenkamp, Institut für Physik, Carl von Barbara Knowles, Jackson Labs, Bar Harbor, Maine, USA Ossietzky Universität Oldenburg, Germany Screening of mutant mice for deficiencies in the Auditory evoked Fields of Melody Processing visual system T. Spieker, Softimage Systems, USA Schmelz Long-term time lapse imaging of axons in histotypic Eric Torebjörk, Dept. of Clinical Neurophysiology, context University of Uppsala , Sweden Microneurography in volunteers and pain patients Rappold Elen Jorum, Dept. of Neurology, University of Oslo, Thomas Holstein, Zoologisches Institut, Universität Norway Heidelberg, Germany Microneurography in volunteers and pain patients Dynamics of MEGAP in Xenopus neuronal growth cones R.A. Meyer, Dept. of Neurosurgery, Johns Hopkins University, Baltimore, USA Stefan Offermanns, Institut für Pharmakologie, Single fiber recordings in the pig Universität Heidelberg, Germany Role of the MEGAP-mDia interaction on actin dyna- Robert H. LaMotte, Dept. of Anesthesiology, Yale mics University, USA Pruritus models G. Elisabeth Pollerberg, Zoologisches Institut, Universität Heidelberg, Germany David C. Yeomans/ Martin S. Angst, Dept. of Anesthe- Role of MEGAP in Retinal Projections siology, Stanford University, USA Microdialysis of skin inflammation

29 Collaborations

Torsten Gordh, Dept.of Anesthesiology, University of Maebashi, Japan, Uppsala, Sweden Presynaptic vesicle traffic Detection methods for pro-NGF Grigori Enikolopov, Cold Spring Harbor Laboratories Per-Anders Jansson, Lundberg Lab., University of Cold Spring Harbor, USA Gothenburg, Sweden NO-signalling in Drosophila Measurement of tissue insulin by microdialysis Armin Huber, Universität Karlsruhe, Germany, Frank L. Rice, Albany Medical College, USA TRP-signalling in Drosophila Analysis of skin biopsies in chronic pain patients Martin Heisenberg/ Bertram Gerber, Universität Würzburg, Germany, Schratt Learning & Memory in Drosophila Michael E. Greenberg, Children´s Hospital, Boston, Heinrich Betz/ Bertram Schmitt, MPI für Hirnfor- USA schung, Frankfurt, Germany, The role of MEF2 in transcriptional regulation of mi- Drosophila NMDA receptors croRNAs Hilmar Bading, Institut für Neurobiologie, Universität Javier Martinez, Institut für Molekulare Biotechnologie, Heidelberg, Germany, Wien, Austria Nuclear calcium signalling in Drosophila The role of miR-138 in dendritic spine development Andreas Draguhn, Institut für Physiologie und Patho- Markus S. Kauppinen, University of Copenhagen, physiologie, Universität Heidelberg, Germany, Denmark Planned work on presynaptic quantal size regulati- LNA technology for the study of neuronal micro on RNAs Peter, Seeburg, MPI für Medizinische Forschung, Schütz Heidelberg, Germany Ingrid Grummt, DKFZ, Heidelberg, Germany In vivo analysis of neuronal microRNAs using AAV Nucleolar disruption by inactivation of the RNA poly- Kerry, Tucker, Institut für Neuroanatomie und merase I transcription factor TIF-IA leads to cell cycle Zellbiologie, Universität Heidelberg, Germany arrest and p53-mediated apoptosis The role of microRNAs in axon development Herrmann-Josef Gröne, DKFZ, Heidelberg, Germany Andreas Draguhn, Institut für Physiologie und Histopathological analysis of nuclear receptor Pathophysiologie, Universität Heidelberg, Germany mouse mutants Electrophysiological characterization of microRNA Peter Krammer, DKFZ, Heidelberg, Germany phenotypes Histopathological analysis of nuclear receptor mouse Herbert Waldmann, MPI für Molekulare Physiologie, mutants Dortmund, Germany Peter Lichter, DKFZ, Heidelberg,Germany APT1 function in spine development Nuclear receptor tailless allows to distinguish neural Alfred Nordheim, Universität Tübingen, Germany stem cells from progenitors in the adult subventricu- SRF regulated microRNAs in the brain lar zone Peter Gass, Central Institute for Mental Health (ZI), Schuster Mannheim, Germany Yoshiaki Kidokoro, Gunma University Medical School, Analysis of learning and memory in glucocorticoid receptor for mutants

30 Collaborations

Norbert Gretz, Center of Medical Research, Universität Nancy J. Rothwell, University of Manchester, UK Mannheim, Germany Inflammation in stroke Affymetrix-based analysis of gene expression Shizuo Akira, Osaka University, Japan Stefan Offermanns, Institut für Pharmakologie, Gene regulation of NF-kB Universität Heidelberg, Germany Clemens Sommer, Universität Mainz, Germany Gq/11-family G-proteins are required for endocanna- TWEAK in cerebral ischemia binoid-mediated neuroprotection Moritz Rossner, MPI für Experimentelle Medizin, Alfred Nordheim, Institut für Zellbiology, Universität Göttingen, Germany Tübingen, Germany Neurogenesis Neuronal migration in the murine rostral migratory stream requires SRF Seeburg Hans-Peter Lipp, Institute of Anatomy, University of Øivind Vidar Jensen, Dept. of Neurophysiology, Zürich, Switzerland University of Oslo, Norway Loss of the limbic mineralocorticoid receptor impairs LTP Expression behavioral plasticity Leszek Kaczmarek, Nencki Institute of Experimental Simon Biology, Warsaw, Poland Andreas Draguhn, Institut für Physiologie and Patho- The role of CREB and CREM in neuronal survival, sy- physiologie, Universität Heidelberg, Germany naptic plasticity and drug addiction – A combined K-ATP channels in MN9D cells molecular genetic and neurobiological approach. Maria Grazia Spillantini, Centre for Brain Repair, Nancy Hynes, Friedrich Miescher Institute, Basel, University of Cambridge, UK Switzerland Regulation of α-synuclein by the engrailed transcrip- β1 integrins regulate mammary gland proliferation tion factors and maintain the integrity of mammary luminal al- veoli. Sang-Hun Lee, Dept. of Biochemistry and Molecular Biology, College of Medicine, Hanyang University, David Ginty, Dept. of Neuroscience, Johns Hopkins South Korea University, Baltimore, USA Regulation of stem cell differentiation by the en- Distinct requirements for SRF and CREB in neuronal grailed genes survival and synaptic plasticity Thomas Perlmann, Ludwig Institute for Cancer Ute Moll, Stony Brook University, New York, USA Research, Karolinska Institute, Sweden p53 expression in mitochondria Nurr1 and the dopaminergic projection to the basal Allan Herbison, Centre for Neuroendocrinology/ Dept. ganglia of Physiology, University of Otago, Dunedin, Joseph Bryan, Dept. of Molecular and Cellular Biology, New Zealand Baylor College of Medicine, USA The role of GABA for GnRH neuron, migration and Loss of midbrain dopaminergic neurons in Sur1 mu- function and the olfactory GnRH system. tant mice

Schwaninger Michael Cleary, Dept. of Pathology, Stanford University School of Medicine, USA Allan Butterfield, University of Kentucky, USA Pbx1 and midbrain dopaminergic neurons ROS in cerebral ischemia

31 Collaborations

Licia Selleri, Dept. of Cell and Developmental Biology, Cornell University Med. School, USA Sprengel Function of Pbx1 in adult mesencephalic dopaminer- Øivind Hvalby/ Vidar Jensen, University of Oslo, Norway gic neurons LTP-Expression Richard Dyck, Dept. of Psychology/ Dept. of Neuro- Andreas Lüthi, Friedrich Miescher Institute, Basel, science, University of Calgary, Canada Switzerland Motor deficiencies in mutant mice Emotional Learning Rüdiger Klein, MPI für Neurobiologie, Martinsried, Carolina Eva, University of Turino, Italy Germany Function of NPY Receptors ErbB4 and the midbrain dopaminergic neurons Steven Petrou, University of Melbourne, Australia Cairine Logan, Dept. of Cell Biology and Anatomy, Epileptic activities in GABA-A receptor mutant mice University of Calgary, Canada Engrailed dependent survival of midbrain dopa- Esa Korpi, University of Helsinki, Finland minergic neurons GluRs in addiction Ulrich Misgeld, Institut für Physiologie and Pathophy- Tucker siologie, Universität Heidelberg, Germany Glucose sensing by KATP channels in mesencephalic Hilmar Bading, Institut für Neurobiologie, Universität dopaminergic neurons Heidelberg, Germany CaMKIV and nucleocytoplasmic HDAC shuttling Söllner Norikazu Nishino, Kyushu Institute of Technology, Jan Behrends, Institut für Physiologie, Universität Kitakushu, Japan Freiburg, Germany Novel synthetic inhibitors of HDAC function Electrophysiology Kirk Mykytyn, Dept. of Pharmacology, Ohio State Bernd Simon, EMBL, Heidelberg, Germany University, Columbus, USA NMR analysis Examination of brain defects in BBS4 knockout mice John Briggs, EMBL, Heidelberg, Germany Bradley Yoder, Dept. of Cell Biology, University of Cryo-electronmicroscopy Alabama, Birmingham, USA Analysis of brain development in floxed Ift88 knock- Spors out mice Amiram Grinvald, Weizmann Institute of Science, Israel Reha Erzurumlu, UMB School of Medicine, Baltimore, Voltage Sensitive Dye Imaging USA Analysis of the barrel cortex in mouse mutants ex- Adi Mizrahi, Hebrew University, Jerusalem, Israel pressing modulators of Rho GTPase function Epilepsy Thomas Theil, University of Edinburgh, Edinburgh, UK Steven Petrou, University of Melbourne, Australia In situ analysis of brain development in the cobble- Epilepsy stone mouse mutant Andreas Schaefer, University College London, England Stefan Wölfl, Institut für Pharmazie und Molekulare Olfactory Behavior Biotechnologie, Universität Heidelberg, Germany Matt Wachowiak, Boston University, USA Biochemical analysis of the cobblestone mouse Calcium Imaging of Odor Responses mutant

32 Collaborations

Joachim Spatz, MPI für Metallforschung, Stuttgart, Ludwig Aigner, Universität Regensburg, Germany Germany Embryonic analysis of mice expressing fluorescent Scanning electron microscopy of organotypic slice proteins under the control of the doublecortin cultures and ciliary defects in the brains of cobbles- promoter tone mutant mice Michael Sporn, Dartmouth Medical School, Hanover, Kurt Gottmann, Universität Düsseldorf, Germany USA EGFP transgenesis of mutant ES cells via lentivirus Triterpenoid compounds and their effect on neuro- Peter Hortschansky, Leibniz Institute for Natural genesis Product Research and Infection Biology, Jena, Germany Unsicker Recombinant BMPs and BMP inhibitors Liliana Minichiello, EMBL Moterotondo, Italy Francesca Ciccolini, Institut für Neurobiologie, Involvement of CEPBs in brain functions Universität Heidelberg, Germany Chaya Kalcheim, Hebrew University Jerusalem, Israel FACS sorting of neurons and stem cells from embry- Development of the SA cell lineage from the NC onic mouse brain Beata Legutko, Polish Academy of Sciences, Krakov, Sangeeta Chawla, Dept. of Pharmacology, University Poland of Cambridge, Cambridge, UK Role of FGF-2 in depression Nucleocytoplasmic shuttling of HDAC5 and its effect Mart Saarma, Biocenter, University of Helsinki, Finland upon neurogenesis GDNF signaling Stefan Wölfl, Institut für Pharmazie und Molekulare Juha Partanen, Biocenter, University of Helsinki, Biotechnologie, Universität Heidelberg, Germany Finland Affymetrix microarray analysis of cortical and stria FGFRs in glial development tal neurogenesis David Kaplan, Montreal Neurological Institute, Canada Stefan Wölfl, Institut für Pharmazie und Molekulare ERK and neuron death Biotechnologie, Universität Heidelberg, Germany Molecular dissection of class II HDAC shuttling and Kerstin Krieglstein, Forschungszentrum Molekular- its role in neurogenesis physiologie des Gehirns (CMPB), Göttingen, Germany Steven Dooley, Medizinische Klinik II, Universitätsklini- Neural functions of TGF-ß kum Mannheim, Germany Adenovirus expressing reporters for and modifiers of Michael Sendtner, Institut für Neurologie, Universität BMP signaling Würzburg, Germany GDF-15 KO analysis Klaus Unsicker, Institut für Neuroanatomie und Zellbiologie, Universität Heidelberg, Germany Rudolf Martini/ Klaus Toyka, Institut für Neurologie, Transmission electron microscopy of organotypic Universität Würzburg, Germany slice cultures for the imaging of peripheral nerve de- GDF-15 and myelination velopment Rüdiger Klein, MPI für Neurobiologie, Martinsried, Peter Wieacker, Institut für Humangenetik, Universitäts- Germany klinikum Magdeburg, Germany Generation of GDF-15 knockout Transcript analysis of fibroblast cultures from pseu- dotrisomy 13 patients Wick

33 Collaborations

Michael Weller, Neurology, Tübingen, Germany/ Clemens Gabher, KIT Karlsruhe, Zürich, Switzerland Macrophage migration TGF-β projects Ruggero Pardi, Milan, Wilfried Roth, Institut für Pathologie, Universität Molecular and cell biological aspects of macrophage mig- Heidelberg, Germany ration BCL-xL Wittum Martina Schnölzer, Proteomics Group, DKFZ, Germany Hypoxia candidate validation and characterization Hilmar Bading, Institut für Neurobiologie, Universität Heidelberg, Germany Guido Reifenberger, Institut für Neuropathologie, DMSPiN Universität Düsseldorf, Germany NOA-04 Andreas Draguhn, Institut für Physiologie und Patho- physiologie, Universität Heidelberg, Germany Andreas von Deimling, Institut für Neuropathologie, DMSPiN Universität Heidelberg, Germany NOA-04, MGMT, brain tumor stem cells Hannah Monyer, Institut für Neurobiologie, Universität Heidelberg, Germany Anthony Ho, Innere Medizin V, Universtität Heidelberg, DMSPiN Germany Stem cell projects Christoph Schuster, Institut für Neurobiologie, Universität Heidelberg, Germany Wittbrodt DMSPiN Manfred Schartl, Biozentrum Uni Wuerburg, Roger Traub, SUNY Health Center, New York, USA Mutagenesis screen Humboldt Juan-Ramon Martinez-Morales, University Sevilla, Witzemann Eye morphogenesis Joseph McArdle, New Jersey Medical School-UMDNJ, Felix Loosli, KIT, Karlsruhe Institute of Technology, Retinal polarity and growth USA AChR subunit switch Paoloa Bovolenta, CSIC, Instituto Cajal, Madrid, Spain, Eye development Daniel Hantai, Institut de Myologie, Hôpital de la Salpétriêre, Paris, France Jean-Stephane Joly, Gif-sur-Yvette, Fance, MuSK-induced CMS Transcriptional control of cell proliferation Rüdiger Rudolf, Forschungszentrum Karlsruhe in der Yunhan Hong, NUS, Singapore, Retinal stem cells Helmholtz-Gemeinschaft, Karlsruhe, Germany AChR subunit trafficking Yijun Ruan, GIS, Singapore, Chip-Seq approaches downstream of Six3 Ernst Stelzer, EMBL, Heidelberg, Light sheet microscopy in development and physiology Urban Liebel, KIT Karlsruhe, High content imaging of embryos in genetic screens

34 Research Profiles

35 Detlev Arendt Current Research

The evolution of the animal central nervous The molecular comparison system of neuron types We combine morphological Research Summary and molecular approaches in We are intrigued by one of the remaining great mysteries a novel evo-devo approach, in animal evolution: how did our central nervous system the molecular comparison of come into existence? What did it look like at first and how neuron types. Animal central did it function? We are especially interested in the CNS of nervous systems are made an extinct animal known as Urbilateria, the last common up of different types of sen- ancestor of humans, flies and most other ‘higher’ animals sory neurons, interneurons, that live today. motor and neurosecretory cells as well as support and glial cells. Each type displays a characteristic “molecular Curriculum Vitae fingerprint”, which is the unique combination of specify- Degrees: 1995 M.Sc. ing transcription factors and downstream effector genes 1998 Ph.D. such as receptors, transmitters or neuropeptides that are 1995-1998: Thesis work under supervision of K. Nübler-Jung: expressed in these cells. The molecular fingerprint is a con- “Comparison of central nervous systems of servative trait that can be retained over more than 600 mil- arthropods, annelids and chordates“ lions of years and allows the tracing of cell types through 1998-2002: Postdoctoral fellow at the EMBL, Heidelberg animal evolution (Arendt, 2004; Arendt et al., 2004; Arendt, 2002-2006: Team Leader 2005; Denes and Jekely et al., 2007; Tessmar-Raible et al., Developmental Biology Unit EMBL, Heidelberg 2006-2007: Group Leader 2007). Fig. 1 depicts our current working hypothesis on ho- Developmental Biology Unit EMBL, Heidelberg mologous cell types shared between the polychaete and since 2007: Group Leader and Senior Scientist vertebrate forebrain, as a summary of the past years’ work. Developmental Biology Unit EMBL, Heidelberg Molecular fingerprinting with cellular resolution is greatly facilitated by the novel Wholemount reflection CLSM (see Contact above). Our molecular dissection of Platynereis develop- EMBL ment has been complemented by ultrastructural studies Developmental Biology Unit on cellular morphologies carried out in collaboration with Meyerhofstrasse 1 G. Purschke (Osnabrück) and H. Hausen (Berlin). 69117 Heidelberg Germany Molecular dissection of Platynereis eye and photorecep- 9 +49-6221-387624 tor development E +49-6221-387166 J [email protected] Three pairs of eyes and three pairs of chemo-/mechano- O http://www-db.embl.de/jss/EmblGroupsHD/g_172.html sensory organs connect to the Platynereis brain (e.g., Arendt et al., 2002). Ongoing and future projects aim at the further molecular and functional characterisation of the Platynereis head sensory cell types, in order to trace their evolution throughout Bilateria. We have shown that the Platynereis brain harbours ciliary photoreceptor cells that share a common “molecular fingerprint” with the rods and

36 Detlev Arendt cones of the vertebrate retina (Arendt et al., 2004). For ex- thus resemble Darwin‘s ‚proto-eyes‘, considered to be the ample, we discovered a vertebrate-type Platynereis Opsin first eyes to appear in animal evolution. These eyespots expressed in these cells (violet in Fig. 1). This is strong indi- cannot form images but enable the animal to sense the di- cation that rods and cones have evolved from a precursor rection of light. They are characteristic for the zooplankton population of deep brain ciliary photoreceptors that exis- larvae of marine invertebrates and are thought to mediate ted already in the last common ancestors of the bilaterians, larval swimming towards the light. Phototaxis of inverteb- the Urbilateria. rate larvae contributes to the vertical migration of marine plankton5, which is thought to represent the biggest bio- mass transport on Earth. We have recently shown how simple eyespots in marine zooplankton mediate phototactic swimming, using the marine annelid Platynereis dumerilii as a model.

Fig. 2: Computer simulation of Platynereis phototaxis Sensory-neuroendocrine cell types in the Platynereis brain To elucidate the evolution of neurosecretory centres in bi- Fig. 1: Homologous cell types in the Platynereis larval episphere and in laterian brains, we molecularly characterise and compare the vertebrate prosencephalon. Violet: ciliary photoreceptor cells; yellow: early differentiating neuron types in the nk2.1-positive rhabdomeric photoreceptor cells/ retinal ganglion cells; green: molecular medial forebrain region in Platynereis and in the zebrafish clock cells. Red: Vasotocin-secreting extraocular photoreceptors. (Tessmar-Raible et al., 2007) At the current state of analysis, Blue: serotonergic cells. Brown: FMRFamidergic chemosensory cells. ae: adult eye, cPRC: ciliary photoreceptor cell, le: larval eye, nsc: nucleus we identified two conserved cell types by molecular fin- suprachiasmaticus, pin: pineal, ret: retina, RGC: retinal ganglion cells, gerprint comparison rPRC: rhabdomeric photoreceptor cells First, we characterized a population of cells in Platynereis that co-express the transcription factors nk2.1, otp and rx, We are also investigating the larval eyes of the Platynereis a vasotocin (vasopressin/oxytocin) - neurophysin prohor- trochophora larva. These are of special importance for mone, and ciliary opsin, and discovered cells of identical phylogenetic comparisons because very similar larval eyes “molecular fingerprint” in fish. This indicates the presence also exist in basal Deuterostomia such as enteropneusts. of vasotocinergic deep brain photoreceptor cells in both We propose that such larval eyes existed already in Urbi- species. Second, we identified a population of flask-shaped, lateria. sensory-neurosecretory cells in the Platynereis brain that Platynereis larval eyes are eyespots composed of two cells co-express the transcription factor otp and resemble a only: a photoreceptor and a shading pigment cell. They subset of vertebrate central-spinal-fluid-contacting neu-

37 Detlev Arendt rons by cytoarchitecture, and by RFamide neuropeptide content. Corroborating homology of the vasotocinergic and FMRFamidergic cell types across Bilateria we find that in both Platynereis and fish they form part of a small population of cells in the developing brain that is demarcated early on by the specific expression of the conserved mi- croRNA miR-7.

Motor-, inter, and sensory neurons of the ventral nerve cord The larval polychaete trunk comprises four larval segments. Each segment harbours motor-, inter-, and sensory neurons that control larval swimming. We have started to determine the molecular fingerprint and the mediolateral distribu- Fig. 3: Conservation of miR-7 expression in early differentiating vasotocinergic and FMRFamidergic cells. MiR-7 is expressed in a restricted popu- tion of these neuron types at larval stages, assigning them lation of early differentiating neurons in the developing Platynereis brain to the different mediolateral progenitor domains as outli- (A). Among these are the vasotocinergic cells (B, white arrows) and the ned above (Denes et al., 2007). For example, we found that FMRFamidergic cells (C, white arrowhead, cf. to 15C), as deduced from hb+ cholinergic somatic motor neurons emerge from the spatially correlating the position of these cells to the axonal scaffold (green). A miR-7+ cell population also exists in the developing fish brain Px6+, nk6+ progenitor domain in Platynereis as they do in (D), including also vasotocinergic (E) and FMRFamidergic (F,G) cells. vertebrates. We validated by cholinesterase staining that these neurons directly innervate the somatic musculature also in Platynereis and that contractions are abolished by Top publications acetyl choline receptor antagonists. We further determined that serotonergic neurons emerge • Arendt, D. (2003). Evolution of eyes and photoreceptor cell ty- from the medial nk2.2 column and sensory neurons from pes. Int. J. Dev. Biol. 47, 563-71. the lateral msx+ domains in polychaete and vertebrate. • Arendt, D., et al. (2004). Ciliary photoreceptors with vertebra- Altogether, these findings are consistent with an overall te-type opsins in an invertebrate brain. Science, 306, 869-871. • Arendt, D. (2005). Genes and homology in nervous system conserved mediolateral architecture of the polychaete evolution: comparing gene functions, expression patterns, and vertebrate trunk CNS, and thus with a common evolu- and cell type molecular fingerprints. Theor. Biosci. 124, 185- tionary origin of nervous system centralisation. This would 197. imply modification of the mediolateral architecture in the • Raible, F., et al. (2005). Vertebrate-type intron-rich genes in the fly and in the nematode. marine annelid Platynereis dumerilii. Science 310, 1325-6. • Steinmetz, P. R. H., et al. (2007). Polychaete trunk neuroecto- derm converges and extends by mediolateral cell intercalati- Structure of the Group on. Proc. Natl. Acad. Sci. 104(8), 2727-32. • Denes, A., et al. (2007). Molecular architecture of annelid ner- Group Leader: Detlev Arendt ve cord supports a common origin of nervous system centrali- Postdoctoral fellows: Peter Hantz, Florian Raible, sation in Bilateria. Cell, 129, 277-288. Kristin Tessmar-Raible • Tessmar-Raible, K., et al. (2007). Evolution of the vertebrate PhD students: Foteini Christodoulou, Alexandru Denes, hypothalamus: An ancient set of sensory- neurosecretory cell Antje Fischer, Keren Guy, Raju Tomer types in the polychaete and vertebrate medial forebrain. Cell, Undergraduates: Benjamin Backfisch, Carmen Döring, 129, 1389-400. Nicola Kegel, Katharina Willmann • Jékely, G., et al. (2008) Mechanism of phototaxis in marine zoo- Technicians: Heidi Snyman plankton. Nature, 456:395-9.

38 Current Research Hilmar Bading Nuclear calcium signaling in the dialogue Changes in the concentration of intracellular calcium as between synapse and nucleus: a result of synaptic activity Role in neuronal survival and memory control virtually all adaptive responses in the adult ner- Research Summary vous system. Calcium activa- Synaptic activity-induced changes in the concentration of tes mechanisms that affect intracellular calcium control many processes in neurons synaptic connectivity, regu- including gene expression. Calcium signals propagating late learning and memory, towards the cell soma and the nucleus are key mediators promote survival, modulate of synapse-to-nucleus communication. Nuclear calcium pain, or cause cell death. Most activity-induced adapta- controls CREB/CBP-dependent transcription and regulations are initiated by synaptic NMDA receptors and require tes genomic programs critical for neuronal survival, plas- for their maintenance signal-induced changes in gene ex- ticity, and memory. pression. For synapse-to-nucleus communication neurons exploit the spatial and temporal diversity of calcium tran- Curriculum Vitae sients associated with electrical activation. Transcriptional Degrees: 1984 MD, MPI for Medical Research, Heidelberg responses depend on how calcium enters the neurons, the and University of Heidelberg amplitude of the signal, how long it lasts and what subcel- 1985-1989: Postdoctoral fellow, lular compartment it invades. One means of conveying a MPI for Molecular Genetics, Berlin signal to the nucleus involves ERK-MAP kinases that trans- 1989-1992: Postdoctoral fellow, Dept of Microbiology and locate to the nucleus and stimulate transcription factors Molecular Genetics, Harvard Medical School, upon being activated by a calcium micro-domain in the Boston, USA immediate vicinity of the site of calcium entry. However, 1993-2001: Group Leader at the MRC Laboratory of Molecular Biology, Cambridge, UK the principal mediator in the dialogue between the syn- since 2001: Professor, Chair of Neurobiology, apse and the nucleus is calcium itself. Synaptic activity and Director, Neurobiology, NMDA receptor stimulation can initiate calcium transients IZN, University of Heidelberg that propagate towards the cell soma and enter the cell nucleus. Contact Department of Neurobiology Nuclear calcium: universal signal in neuronal survival IZN and long-term memory University of Heidelberg Nuclear calcium signaling controls the expression of a wide Im Neuenheimer Feld 364 69120 Heidelberg variety of target genes, primarily by stimulating CREB/CBP- Germany mediated transcription. Our hypothesis is that nuclear cal- 9 +49-6221-548218 cium acts as a universal signal for persistent adaptations E +49-6221-546700 in the nervous system. To test this hypothesis, we have fo- J [email protected] cused on two adaptive processes: activity-dependent neu- O http://izn.uni-hd.de/ ronal survival (i.e. acquired neuroprotection) and memory formation. Using tools to interfere selectively with calcium signaling in the nucleus of hippocampal neurons, we

39 Hilmar Bading have been able to demonstrate that the neuroprotection afforded by action potential bursting and synaptic NMDA receptor activation is indeed dependent upon nuclear calcium signaling. Whole genome transcriptional profiling in hippocampal neurons revealed a nuclear calcium-regulated genomic survival program consisting of about a do- zen genes. The neuroprotective activity of some of these genes has been demonstrated in vitro as well as in in vivo models of neurodegeneration. The role of nuclear calcium in learning and memory is being analyzed in the fruit fly Drosophila melanogaster. The results obtained so far indicate that the formation of long-term memory following associative olfactory learning is impaired in transgenic flies expressing an inhibitor of nuclear calcium signaling.

Fig. 2: Organotypic hippocampal slice culture containing a neuron expressing green fluorescent protein. The cell nuclei are stained with Hoechst. Image by Daniela Mauceri.

are using stereotaxic delivery of recombinant adeno-associated viruses to express recombinant calcium indicators targeted to the cell nucleus in the rodent brain. Techniques of imaging calcium signals in vivo are being developed. We have also generated transgenic Drosophila melanogaster expressing a recombinant nuclear calcium indicator in the

Fig. 1: Illustration of a neuron containing a highly infolded nucleus. The geometry of the nucleus was obtained using three-dimensional image reconstruction based on confocal microscopy data. The surface plot illustrates a nuclear calcium signal evoked by a burst of action potential firing. Picture by Anja Eder and Gillian Queisser.

Nuclear calcium imaging in vivo A major challenge for the future is to develop tools and technologies to detect nuclear calcium signals in vivo. Gi- ven the central role of nuclear calcium in synaptic plastici- Fig. 3: Differential signaling by synaptic and extrasynaptic NMDA recep- ty-related gene expression, we are particularly interested tors. Schematic illustration of calcium-regulated pathways controlling in studying nuclear calcium transients during learning. We survival and memory. Illustration by Oliver Dick and Malte Wittmann.

40 Hilmar Bading nervous system. These flies are being used to monitor nuc- Top publications lear calcium signals during associative olfactory learning. • Vanhoutte, P., and Bading, H. (2003). Opposing roles of synaptic and extrasynaptic NMDA receptors in neuronal calcium Extrasynaptic NMDA receptor signaling: CREB shut-off signalling and BDNF gene regulation. Curr. Opin. Neurobiol. and cell death pathways 13, 366-371. The transcription-promoting activities of synaptic NMDA • Arnold, F., et al. (2005). Microelectrode array recordings of cultured hippocampal networks reveal a simple model for receptor-induced nuclear calcium signals are antagonized transcription and protein synthesis-dependent plasticity. J. by a calcium signaling pathway that is initiated by calci- Physiol. 564 (Pt 1), 3-19. um flux through NMDA receptors located outside synaptic • Papadia, S., et al. (2005). Nuclear calcium and CREB family me- contacts. Extrasynaptic NMDA receptors couple to a CREB diate a late-phase of activity-dependent neuroprotection. J. shut-off pathway and cause cell death. Thus, the decision Neurosci. 25, 4279-4287. whether a neuron survives (and perhaps undergoes plasti- • Soriano, F.X., et al. (2006). Preconditioning doses of NMDA city) or dies after glutamate exposure is dependent on the promote neuroprotection by enhancing neuronal excitability. location of the NMDA receptor activated. This concept of J. Neurosci. 26, 4509-4518. differential signaling by synaptic and extrasynaptic NMDA • Eder, A., and Bading, H. (2007). Calcium signals can freely cross the nuclear envelope in hippocampal neurons: Somatic calci- receptors has wide-ranging implications, especially for um increases generate nuclear calcium transients. BMC Neu- the understanding and treatment of neuro-pathological rosci. 8, 57 conditions such as stroke in which brain damage may be • Wiegert, J.S., et al. (2007). Diffusion and Not Active Transport caused by the stimulation of extrasynaptic NMDA recep- Underlies and Limits ERK1/2 Synapse-to-Nucleus Signaling in tors. Hippocampal Neurons. J. Biol. Chem. 282, 29621-29633 • Zhang, S.J., et al. (2007). Decoding NMDA receptor signaling: identification of genomic programs specifying neuronal survival and death. Neuron 53, 549-562. • Wahl, A.-S., et al. (2009). Hypoxic/ischemic conditions induce expression of the putative pro-death gene Clca1 via activation of extrasynaptic N-Methyl-D-Aspartate Receptors. Neuros- ci. 158, 344-352

Structure of the Group Group Leader: Hilmar Bading Postdoctoral fellows: C. Peter Bengtson, Otto Bräunling, Bettina Buchthal, Oliver Dick, Anja Eder, Anna Hagenston, Daniela Mauceri, Ana Oliveira, Jan-Marek Weislogel, Shengjia Zhang PhD students: Désirée Ditzel, Eckehard Freitag, David Lau, Li Lu, Anna-Sophia Wahl, Simon Wiegert Undergraduates: Martina Bujard, Jana Schlüter, Philip Tröster, Yan Yu, Ming Zou Fig. 4: 3D image reconstruction of nuclei from hippocampal neurons. Technicians: Iris Bünzli-Ehret, Andrea Hellwig, Picture by Gillian Queisser and Malte Wittmann. Ruth Jelinek, Ursula Weiss

41 Dusan Bartsch Current Research Molecular and cellular basis of normal and Role of voltage gated pathologic cognition, learning and memory, calcium channels in and mental retardation learning and memory Our focus is the role of Research Summary LVGCCs in learning and We study normal and pathological learning and memory in memory, memory extinction molecular terms. Although the role of genes in basic synaptic and age dependent memory functions can be studied in cultured neurons, the role of genes in loss. Since no subunit- cognition can be studied only in the functional brain. Therefore, specific antagonists or we create targeted and regulated genetic modifications of agonists of LVGCCs are candidate genes specifically in the areas of the rodent brain available, genetic tools are necessary to dissect the specific involved in learning and memory. The genetically modified mice role of Cav1.2 and Cav1.3 in normal and pathologic brain and rats are then studied using current methods of molecular functions. We have generated mice with conditional Cav1.3 biology, electrophysiology, behavior and pharmacology. alleles. By breeding them with mice with tissue-specific expression of tamoxifen-inducible Cre recombinase, we Curriculum Vitae are able to study the roles of Cav1.3 in defined tissues at Degrees: 1984 Ph.D. in Biology, Charles University, specific time-points, thus allowing to differentiate the Praha, Czech Republic role of Cav1.3 in developing, adult and aging organs. We 1979-1983: B.S., Molecular Biology and Virology, have generated and tested forebrain and hippocampus- Charles University, Praha specific promoters driving the Cre recombinase, enabling 1983-1984: PhD, Biology, Charles University, Praha 1984-1987: Staff scientist, Dept. of Exp. Virology, Institute for us to study the role of Cav1.3 in hippocampus-dependent Sera and Vaccines, Praha learning and memory. Recently, we were able to target 1987-1988: Guest scientist, German Cancer Research Center sub-hippocampal regions. Such genetic modifications (DKFZ), Heidelberg, Germany allow us to study the role of Cav1.3 in brain and other 1988-1992: Postdoctoral fellow, DKFZ selected tissues and generate mouse models for specific 1992-1993: Postdoctoral research fellow, HHMI and role of LVGCCs in diverse physiological functions. Center for Neurobiology & Behavior, Columbia University, New York, USA Role of MEGAP in mental retardation 1993-2000: Research associate scientist, Columbia University, New York Mental retardation is a common condition affecting since 2000 Professor of Molecular Biology at University between 0.3-3% of the population, depending on severity. of Heidelberg and Central Institute of Mental We have generated mice with mutations mimicking a Health, Mannheim, Germany recently identified mutation in a novel RhoGAP protein family (MEGAP) found in a patient with severe mental Contact retardation (Endris et al.; 2002). We generated mice with Department of Molecular Biology stop codon in exon 3 of Megap gene, resulting in premature Central Institute of Mental Health termination of MEGAP protein translation. Megap -/-mice J5, 68159 Mannheim, Germany develop hydrocephalus and show defects in neuronal 9 +49-621-1703 6202 morphology. On the behavioral level, Megap -/-mice E +49-621- 1703 6205 demonstrate increase in anxiety. Molecular studies with J [email protected] O http:// www.zi-mannheim.de Megap -/- mice indicate that the ERK1 signaling pathway is significantly affected in Megap knockout mice.

42 Dusan Bartsch

Modeling serotonergic defects in depression using conditional regulation of serotonin concentration in transgenic mice The serotonergic system is an important modulator of many developmental, behavioral and physiological processes, and its perturbation is a major etiological component of depression. The serotonin transporter (SERT) plays a key role in the regulation of central serotonergic neurotransmission by removing serotonin from the synaptic cleft. Stressful life events in humans are known to be followed by depressive episodes in susceptible individuals. Vulnerability seems to be genetically anchored in the serotonergic system. Tph2 is a key enzyme in serotonin synthesis. We have generated congenic B6.D2-Tph2 C1473G mice carrying the Fig. 1: Expression of Megap mRNA in mouse embryo (E11). Wholemount tryptophan hydroxylase 2 allele (Tph2) of the DBA2 mouse in-situ hybridization with antisense Megap RNA probe. At E11, Megap is strain with the C1473G polymorphism in the defined expressed in the brain and spinal cord. C57BL76 genetic background. The C1473G mutation results in reduced activity of Tph2 enzyme. By combining tetracycline inducible tTA system and CreERT2 mediated B6 and B6.D2-Tph2 C1473G mice with conditional alleles recombination and the miRNA technology for generating of Tph2 and Sert generated in our laboratory, we are able inducible gene knockdowns in the rat brain. to manipulate the serotonin concentration in the mouse By combining the above methods, we hope to manipulate brain either during development or in adult animals. bi-directionally (overexpression and knockdown) gene B6.D2-Tph2C1473G mice show increased anxiety. Currently, expression of rat genes and make rat models accessible to we test the interaction of C1473G polymorphism with reverse genetics. chronic mild stress. This model provides an opportunity to study regulatory pathways specifically affected in Age dependent memory loss vulnerable animals under stressful conditions. Aging is associated with a decline in memory. This impairment is progressive and widespread, affecting Inducible and regulated gene expression in the rat about 40% of people over the age of 65. Animal models brain have been developed to study both the genetic and The recent progress in molecular neuroscience allows first environmental components of age-dependent memory insights in mechanisms of neuronal function on molecular decline. Interestingly, even in inbred strains of rats or mice, and cellular level. On the behavioral level, this has been only a part of the genetically homogenous population to some degree complemented by studies in genetically develops memory deficits with aging, indicating that modified mice. Largely due to the lack of germline forming epigenetic changes, involving changes in gene expression, embryonic stem cells (ES) in rats, major effort to re-establish are the molecular keys to the memory decline. We have many of well studied behavioral, pharmacological and generated genetic tools which allow us to manipulate electrophysiological rat models in mice was undertaken; concentrations of both NMDA receptor and LVGCCs so far with only partial success. Many of the complex selectively in the hippocampus of transgenic rats during behaviors exhibited by rats are not matched by similar mice their lifespan. We have generated transgenic rats with behaviors. We therefore developed molecular tools that the NMDA receptor (NR1) and LVGCC ( Cav1.3) expression allow us to study the role of genes in complex behaviors in controlled reversibly by a tetracycline regulated promoter rats. We explore inducible transgene expression using the (NMDAtet-O and LVGCCtet-O). To target the expression

43 Dusan Bartsch

Top publications • Chen, A., et al. (2003). Inducible enhancement of memory storage and synaptic plasticity in transgenic mice expressing an inhibitor of ATF4 (CREB-2) and C/EBP proteins. Neuron 39, 655-669. • Raich, W.B., et al. (2003). Characterization of Caenorhabditis elegans Homologs of the Down Syndrome Candidate Gene DYRK1A. Genetics 163, 571-580. • Strekalova, T., et al. (2004). Stress-induced anhedonia in mice is associated with deficits in forced swimming and exploration. Neuropsychopharmacol. 29, 2007-2017. • Strekalova, T., et al. (2005). Stress-induced hyperlocomotion as a confounding factor in anxiety and depression models in mice. Beh. Pharmacol. 16, 171-180. • Lee, J.A., et al. (2006). PKA-activated ApAF-ApC/EBP heterodi- mer is a key downstream effector of ApCREB and is necessary and sufficient for the consolidation of long-term facilitation. Fig. 2: Inducible expression of eGFP in the forebrain of transgenic rats. The Journal of cell biology 174, 827-838. Rats with the tetracycline activator driven by CamKII promoter CamKII- tTA) were bred to rats with ptet promoter driven eGFP. Double transgenic • Strekalova, T., et al. (2006). Selective effects of citalopram in a rats (CanKII-tTA x ptet-eGFP) show high expression of eGFP as detected mouse model of stress-induced anhedonia with a control for by anti-GFP antibody. The expression of the reporter is detectable in the chronic stress. Beh. Pharmacol.17, 271-287. forebrain and can be regulated by doxycycline in drinking water. Here, we • Lee, S.H., et al. (2007). Nuclear translocation of CAM-associa- used an insulator LC1 BAC to obtain reproducible and highly inducible ted protein activates transcription for long-term facilitation in expression of the tetO driven transgene. Aplysia. Cell 129, 801-812. of both receptors to the hippocampus, we breed the NMDAtet-O and LVGCCtet-O mice with tTACaMKII rats expressing the tTA activator under control of the CaMKII promoter. To inducibly and reversibly decrease the NMDAR and LVGCC concentration in the neurons, we have developed a tTA based expression system for overexpression of miRNAs targeting both receptors. These genetic tools allow us to perform long-term manipulations of NMDAR and LVGCC concentration in the rat hippocampus. We can thus perform longitudal studies with the same rats over their lifetime and combine behavioral analysis with neuroimaging. Structure of the Group Group Leader: Dusan Bartsch Postdoctoral fellows: Stefan Berger, Kai Schönig, Robert Waltereit, Tillmann Weber PhD students: Vera Beier, Katrin Bartels, Sergej Kutscherjawy, Siri Malmgren Technicians: Ariana Frömmig , Katja Lankisch, Sabine Nescholta, Brigitte Pesold

44 Current Research Martin Bohus Pain perception assessed Pain perception, learning and memory, and by Laser-Evoked Potentials treatment development in patients with and functional MRI in trauma related disorders patients with Borderline Personality Disorder Research Summary Background: Borderline The overarching aim is to improve the current treatments for pa- Personality Disorder (BPD) tients with trauma related disorders (esp. Posttraumatic Stress is a frequent psychiatric dis- Disorder (PTSD), Borderline Personality Disorder (BPD). Our re- order, and pain perception search team is investigating the genenvironment interaction in was shown to be attenuated the development, the central neuropsychological mechanisms in BPD. Our findings from a study using laser-evoked pain of maintenance and the interaction of psychopharmacology potentials suggest that sensory-discriminative pain com- and psychotherapy in its treatment. ponents seem to be unaffected in this patient population and affective-motivational pain components may be al- Curriculum Vitae tered in BPD. Studies in healthy subjects have revealed a Degrees: 1988 MD University of Freiburg pain circuit consisting of thalamus, somatosensory cortex, 2001 Habilitation University of Mannheim 1979-1985: Medical School, University of Freiburg insula, and anterior cingulate cortex (ACC). We used func- 1985-1988: Research scientist, Dept. of tional MRI and heat pain stimuli to localize alterations in Immunology, University of Freiburg pain processing in BPD. 1988-1992: Clinical training (Department of Psychiatry and Neurology, University of Freiburg) Method: Patients with Borderline Personality Disorder ac- 1992: Speciality in Psychiatry and Psychotherapy cording to DSM-IV and healthy controls are investigated 1992-1996: Director of interdisciplinary priority program: (IPP) using brief radiant heat pulses and 7 channel EEG recor- Vulnerability Marker for Major depression 2000: Speciality in Psychosomatic medicine dings. In addition participants underwent functional MRI 2000-2003: Leading scientist of the Freiburg/ Bale Center in the frame during heat pain stimulation. Two stimulus conditions of the Borderline personality disorder research foundation were applied in a randomized fashion: First, a fixed tem- since 2000: Director of IPP: perature was used. Second, a temperature that was percei- Neurobiological aspects of borderline personality disorder 2001: Habilitation for Psychiatry and Psychotherapy ved equally painful by all participants was applied. 2001: Full Professorship at the Central Institute of First results: BPD patients tended to have higher pain Mental Health, Mannheim, (ZI) since 2003: Chair in Psychosomatics and Psychotherapy, thresholds and pain ratings were reduced to 25 % of cont- University of Heidelberg, Director, Dept. of rols. In contrast, mean amplitudes of LEPs in the BPD group Psychosomatics and Psychotherapy, ZI were not reduced compared to those of controls. The abi- since 2003: Associated professorship at the lity to spatially discriminate painful stimuli did not differ University of Arts, Karlsruhe between both groups. In fMRI, all classical pain regions Contact were activated by stimulation with heat pain in BPD patients as well as controls. Group comparison revealed less Department of Psychosomatic Medicine and Psychotherapy Central Institute of Mental Health activation in Somatosensory Association Cortex (Area 7) in J5, 68161 Mannheim, Germany BPD compared to controls under fixed stimulus intensity. A +49-621-1703 4001 During stimulation with equal subjective pain intensities E +49-621- 1703 4005 we found more deactivation in ACC (perigenual cingulate) K [email protected] o http://www.zi-mannheim.de in BPD compared to controls.

45 Martin Bohus

Conclusions: Results of our study are consistent with the idea that the anterior cingulate cortex plays a role in altered pain perception in patients with BPD.

Neuroendocrinological dysregulations in female patients with Borderline Personality Disorder Background: Borderline personality disorder (BPD) is characterized by a long lasting, often chronic pattern of dysfunctions in emotional regulation, interpersonal re- lationships, self-image, and impulse control. The cardinal symptom is a state of aversive tension. Experiencing these symptoms several times a day might lead to a state of chronic stress. Several studies have investigated the physiological consequences of BPD symptoms on neuroendocrine systems. Dysregulations of the hypothalamic-pituitary-adrenal (HPA) axis, however inconclusive, have been found. Methods: In this study, we set out to examine BPD patients in a standardized psychosocial stress paradigm, which is known to lead to substantial increases in HPA axis parameters. Cortisol and catecholamines were measured. A total Fig. 1: Group comparison of responses to a temperature individually ad- of 21 female BPD patients and 21 control subjects current- justed to produce equally perceived pain intensity. Brain activity during ly follows the study protocol. All subjects are medication individually adjusted painful heat stimulation differed between patients with borderline personality disorder (BPD) and controls during the early- free, have a regular menstrual cycle and are taking part in stimulation phase in the left dorsolateral prefrontal cortex and the right the study during their luteal phase. Comorbidity with psy- posterior parietal cortex. In the late phase of individually adjusted heat chiatric disorders like schizophrenia, current depression, pain, intergroup differences in the brain activity were seen in the perige- current drug abuse, and current severe eating disorders is nual part of the anterior cingulated cortex and in the right amygdala. excluded. Results: Preliminary results show that patients with a BPD Impact of Hydrocortisone and Psychotherapy on diagnosis showed similar baseline levels in cortisol and Traumatic Memory Processing in Patients with complex catecholamines as in control subjects. However, in the pa- PTSD tients group a substantial hyporeactivity in HPA axis func- Overall aim of this project: to investigate the role of gluco- tion has been found in comparison to the controls as a corticoids (GC) in traumatic memory retrieval. Animal and reaction to the stressor. Furthermore, sympathetic activity human researches indicate that elevation of central glu- as measured by the catecholamines was higher in patients cocorticoid levels facilitates memory consolidation while prior to the stressor, but no differences have been obser- impairing delayed memory retrieval. Thus, we postulate ved after the stressor. that patients suffering from severe PTSD should benefit Conclusions: The results will be discussed with regard to from hydrocortisone application during exposure based the effects that borderline-typical psychopathology might psychotherapy. have exerted on neuroendocrine function. The project is composed of the following parts: 1. A clinical trial investigates both the efficacy of oral hy-

46 Martin Bohus

Top publications • Tebartz van Elst, L., et al. (2003). Frontolimbic Brain Abnorma- lities in Patients with Borderline Personality Disorder. A Volu- metric MRI Study. Biol. Psychiat. 54 (2), 163-71. • Lieb, K., et al. (2004). Borderline personality disorder. Lancet 364, 453-461. • Schmahl, Ch., et al. (2004). Differential nociceptive deficits in patients with borderline personality disorder and self-injuri- ous behavior: Laser-evoked potentials, spatial discremination of noxious stimuli, and pain ratings. Pain 110, 470-479. • Bohus, M., et al. (2006). Neural correlates of antinociception in borderline personality disorder. Arch. Gen. Psych. 63, 659-667. (first authors contributed equally) • Rüsch, N., et al. (2007). Shame and implicit self-concept in wo- men with borderline personality disorder. American J. Psych- iat. 164, 1-9. • Geuze, E., et al. (2007). Altered Affective and Cognitive Pain Processing in Veterans with Posttraumatic Stress Disorder. Arch. Gen. Psych. 64 (1), 76-85. • Philipsen, A., et al. (2008). Attention deficit/hyperactivity disorder as a potentially aggravating factor in borderline personality disorder. British J. Psychiat. 192, 118-123. • Kraus A., et al. (2008). Amygdala deactivation as a neural cor- Fig. 2: Mean salivary cortisol concentrations during the course of the relate of pain processing in patients with borderline personali- stress induction study in 15 BPD patients and 17 healthy controls. The ty disorder and co-occurrent PTSD. Biol. Psychiat. 65, 819-822. social stressor (TSST) is indicated by grey bars (the narrow grey bar indi- • Kraus, A., et al. (2009. Differentiation of Pain Ratings in Com- cates the introduction to the TSST). The data shown are the mean plus bat-related Posttraumatic Stress Disorder. Pain. 143, 179-185. standard error of mean. • Ebner-Priemer, U.W., et al. (2009). Emotional learning during dissociative states in Borderline Personality Disorder. J Psychi- drocortisone and dialectical cognitive therapy (DCT) at. and Neurosci. 34, 214-222. and the combination of both in the retrieval of traumatic memories and overall symptomatology in patients suffering from severe chronic PTSD. 2. In order to define neurobiological predictors for (early) treatment response, HPA - axis function will be assessed in all patients prior to treatment. Structure of the Group 3. To study the central mechanisms of glucocorticoid-mediated traumatic memory processing, individualized Group Leader: Martin Bohus trauma related scripts will be presented during fMRI Senior Scientist: Christian Schmahl and modified by either oral hydrocortisone or the glu- Postdoctoral fellows: Uli Ebner-Priemer, Nicolaus Kleindienst, Petra Ludäscher, Jana Mauchnik cocorticoid- antagonist mifepristone (misoprostol). Scientists: Sonia Kiko, Iris Klossika, Petra Ludaescher, Anna Mall, Jana Mauchnik, Matthias Limberger, Joachim Wiskemann, Technicians: Ingar Niedfeldt

47 Francesca Ciccolini Current Research Proliferation and differentiation of neural NSC proliferation and main- stem cells tenance Research Summary During aging the number Multipotent and self-renewing neural stem cells (NSCs) of neural stem cells and in the adult mammalian brain continue to generate their ability to proliferate new neurons throughout the life span of the animal. progressively decrease. The Proliferation of NSCs in the telencephalon is regulated mechanisms underlying by temporal and regional clues, which reflect modulatory these changes are not clear. signalling interaction between the NSCs and their niche. While proliferating stem cells To directly study this regulation we have developed a can generate two different daughter cells by undergoing strategy to isolate neural stem cells from different regions asymmetric cell division or identical progeny by dividing of the embryonic and the adult brain. The ultimate goal symmetrically. In addition, stem cells are capable of self- of our work is to device a strategy to identify neural stem renewing as well as differentiating cell divisions. Symmetric cell behaviour in vivo. self-renewing, asymmetric self-renewing and symmetric differentiating cell divisions will lead respectively to Curriculum Vitae expansion, maintenance and depletion of the stem cell pool. In adult tissues a further layer of control of the stem Degrees: 1994 Ph.D. cell pool homeostasis is represented by the ability of stem 1991-1994: University of Rome la Sapienza, Rome, Italy, and Imperial Cancer Research Fund (ICRF) Tumour cells to undergo transitions between active and quiescent Virus Group (TVG, Department of Pathology, states of proliferation. Therefore, to fully understand how University of Cambridge, UK any factor may affect stem cell proliferation it is not enough 1994-1996: ICFRF, TVG, Department of Pathology, to determine its mitogenic activity but also to consider University of Cambridge, UK how it may affect stem cells self-renewal and whether it 1997-2002: MRC Cambridge Centre for Brain Repair, and acts at the levels of actively proliferating or quiescent stem Laboratory of Molecular Signalling, cells. Babraham Institute, Cambridge, UK since 2002 Group Leader, Department of Neurobiology, IZN, A major obstacle to such studies is represented by the University of Heidelberg fact that due to the absence of specific markers stem cell Contact identification is mostly based on retrospective attribution of stem cell properties. Defining characteristics of NSCs Department of Neurobiology (IZN) are their ability to maintain long-term self-renewal and University of Heidelberg multipotency and they are tested in vivo by means of Im Neuenheimer Feld 364 69120 Heidelberg Germany clonal assays. Reports from several laboratories, including A +49-6221-548696 ours, have shown that Epidermal Growth Factor not E +49-6221- 546700 only is the main mitogen inducing clone formation from K [email protected] perinatal and postnatal adult NSCs but also that levels of o http://izn.uni-heidelberg.de EGF (EGFR) receptor expression specifically increase during development in NSCs and that FGF-2 increases expression EGFR in NSCs. In addition, using a fluorescently tagged EGF and flow cytomentry (Fig. 1). we have isolated cell

48 Francesca Ciccolini populations expressing high levels of EGFR (EGFRhigh) from NSC, probably by modulation of intracellular calcium different regions of the embryonic and adult telencephalon homeostasis. However, due to the problems associated and have shown that they are highly enriched for NSCs. with NSC identification it is not known whether these We are using this direct mean of NSC identification to effects are direct or indirect. It is also not clear how investigate regional and temporal specification of NSCs neural activity interacts with other identified signalling and the mechanisms underlying the effect of growth components of the stem cell niche. We are using calcium factors on NSC proliferation and maintenance. imaging and electrophysiology to functionally characterize the membrane properties of isolated NSCs (Fig. 2). Effect of neural activity on NSC proliferation and This functional characterisation is combined with differentiation the analysis of gene expression in NSCs of relevant neurotransmitters and the effect of their activation on NSC It is well known that the behaviour of NSC is not only function. modulated by growth factors and cell interactions but also by neural activity. Changes in neural activity affect not only the proliferation and differentiation of NSC derived progenitors but they may also regulate the proliferation of the primitive NSCs. A growing body of evidence indicates that non-synaptic release of neurotransmitters such as GABA and glutamate affects the proliferation of

Fig. 2: Photograph of representative EGFRhigh cells taken before whole- cell patch clamp recording at 40x magnification observed with DIC.

Fig. 1: Detection of cells expressing high levels of EGFR in freshly dissociated telencephalic cells. Example of sorting plot obtained after staining with EGF coupled to alexa 488 (E-A488) and Propidium Iodide (PI).

49 Francesca Ciccolini

Top publications • Ciccolini, F., et al. (2003). Local and global spontaneous calcium events regulate neurite outgrowth and onset of GABAer- gic phenotype during neural precursor differentiation. J. Neu- rosci. 23, 103-11. • Ciccolini, F., et al. (2005). A Prospective isolation of late development multipotent precursors whose migration is promoted by EGFR. Dev. Biol. 284, 112-25. • Gakhar-Koppole, N., et al. (2008). Neuronal activity requires so- luble Amyloid Precursor Protein α (sAPP α to promote neurite outgrowth in neural stem cells derived neurons via activation of MAPK pathway. EJN 28, 871-882. • Gakhar-Koppole, N., et al. (2008). Depolarization promotes GAD 65-mediated GABA synthesis by a post-translational mechanism in neural stem cell-derived neurons. EJN 27, 269-83. • Cesetti, T., et al. (2009). Analysis of stem cell lineage progression in the neonatal SVZ identifies EGFR+/NG2- cells as transit- amplifying precursors. Stem Cells. 27, 1443-1454.

Structure of the Group Group Leader: Francesca Ciccolini Postdoctoral fellows: Tiziana Cesetti, Nidhi Gakhar PhD-Students: Kirsten Obernier, Yongjoon Suh Undergraduate: Phillip Hundeshagen Technicians: Gabriele Hölz-Wenig, Claudia Mandl

50 Current Research Andreas von Deimling

Identification of epigene- Molecular genetics of tumours of the cen- tically silenced tumor sup- tral (CNS) and the peripheral nervous system pressor gene candidates in (PNS) glioblastomas. (Dr. W. Mueller) Research Summary Epigenetic silencing of ge- The research focus is on human tumours of the CNS and nes by promoter hyperme- PNS. Lines of investigation include the search for genes thylation constitutes an involved in the initiation or progression of these tumors, alternative way of gene in- mechanisms of epigenetic regulation, mechanisms of activation. In an effort to identify novel tumor suppressor apoptosis, novel functions of the NF1 gene and molecular gene candidates (TSG), we took advantage of the reversi- analyses in preclinical and clinical studies. bility of epigenetic silencing by pharmacological manipulation of cultured gliomas with the substance 5`-aza 2`-de- Curriculum Vitae oxicitidine (5`-aza-dC). Combined with a genome wide, Degrees: 1988 MD in Virology microarray-based gene expression profiling it was possi- 1995 Habilitation ble to identify novel TSG candidates, that were inactivated 1980-1987: Medical School (Freiburg) 1988-1990: Residency (AIP), Neuropathology, Zurich primarily by promoter hypermethylation. The expression 1990-1992: Postdoctoral fellow, Neurogenetics, profile of cells that were not treated with 5`-aza-dC where Massachusetts compared to the expression profile of the same cells fol- General Hospital and Harvard Boston lowing demethylation treatment. Genes that reveal signifi- 1992-1995: Residency (AIP), Neuropathology, Bonn cant up-regulation of their expression after 5`-aza-dC ma- 1995: Board certification Neuropathology nipulation are candidates to be tested in ongoing studies. 1995-1998: Associate Professor, Neuropathology, Bonn Methylation patterns in promoters of RUNX3 and TES have 1998-2006: Full Professor, Neuropathology, Charité Berlin already been determined (see Fig. 1). Candidates with dif- Since 2007: Full Professor, Neuropathology, Heidelberg ferential methylation in glioblastomas may proof to be relevant prognostic or predictive markers such as MGMT or Contact future targets for tailored therapy. Department of Neuropathology Institute for Pathology Molecular diagnostics in phase I, phase II and phase III University of Heidelberg trials. (PD Dr. C. Hartmann) Im Neuenheimer Feld 220/221 69120 Heidelberg We provide molecular diagnostics for the following Germany studies: A +49-6221-562603 E +49-6221-564566 RTC 22033-26033 (Evaluating primary chemotherapy with K [email protected] temozolomide vs. radiotherapy in patients with low grade o http://www.klinikum.uni-heidelberg.de/ gliomas with stratification for genetic 1p/19q loss: a phase neuropathologie.370.0.html III study) is coordinated by the European Organization for Research and Treatment of Cancer (EORTC). The objectives are to compare the progression-free survival of patients with low-grade gliomas treated with radiotherapy versus

51 Andreas v. Deimling

temozolomide, and to compare overall survival, the time to death before second progression in patients treated incidence of late toxicity, toxic effects and the quality of either by Temozolomide or PCV chemotherapy before life of patients treated with these regimens. Patients are radiation therapy or Temozolomide or PCV chemotherapy stratified according to participating centre, chromosome after radiation therapy, and to validate differences in 1p status, contrast enhancement on MRI, age, and WHO outcome of patients with anaplastic oligodendroglial and performance status. Patients are randomized to 1 of 2 astrocytic gliomas in respect to 1p/19q status and MGMT treatment arms. methylation status. Lilly LY317615 (Enzastaurin (LY317615) with concomitant UKT-05 (Radiotherapy and concomitant low-dose radiation therapy, followed by enzastaurin maintenance Indometacin and temozolomide therapy and adjuvant therapy in subjects with newly diagnosed glioblastoma temozolomide therapy (one week on/one week off) - a multicenter, open-label, uncontrolled Phase II study) in newly diagnosed glioblastoma: a phase II study) is is coordinated by the Klinische Kooperationseinheit coordinated by the department for neurology at the Neuroonkologie (DKFZ). The objectives are evaluation of university Tübingen. The objective is to test if patients progression-free survival in patients with newly diagnosed treated by intensified temozolomide chemotherapy glioblastoma without promoter methylation of the in conjunction with the COX inhibitor indometacin MGMT gene treated with enzastaurin, and to investigate during radiation therapy followed by weekly alternating safety and tolerability. NOA-04 (Randomized phase III temozolomide chemotherapy perform better than those trial of sequential radiochemotherapy for anaplastic patients in the chemotherapeutical arm of the EORTC oligodendroglial and astrocytic gliomas WHO grade 26981 study. III using PCV or temozolomide) is coordinated by the GGN-II/CP1 (German Glioma Network/Central Project department for neurology at the university Tübingen. The 1) is coordinated by the Klinische Kooperationseinheit objectives are to determine time to second progression or Neuropathology (DKFZ). Microarray-based profiling will

Fig. 1: Bisulfite sequencing and MSP (methylation sensitive PCR) of RUNX3 and TES in primary cultures of glioma, U87 and controls. Presence or absence of methylated CpGs matches the results of MSP. Black dot - methylated CpG; white dot - unmethylated CpG; M - methylated in MSP assay; U - unmethylated in MSP assay

52 Andreas v. Deimling be performed at the genomic and transcriptional level Top publications to systematically characterize molecular aberrations in • Mueller, W., et al. (2007). Downregulation of RUNX3 and TES by glioblastomas. hypermethylation in glioblastoma. Oncogene 256(4), 583-93. • Holtkamp, N., et al. (2007). MMP-13 and p53 in the progres- Neurofibromatosis type I (D. Reuss) sion of malignant peripheral nerve sheath tumors. Neoplasia 9(8), 671-7. Plexiform neurofibromas in NF1 patients are at risk • Holtkamp, N., et al. (2007). Characterization of the amplicon for transformation to malignant peripheral nerve on chromosomal segment 4q12 in glioblastoma multiforme. sheath tumors (MPNST). It is widely agreed on that the Neurooncology 9(3), 291-7. occurrence of an additional somatic mutation within the • Krex, D., et al. (2007). Long-term survival with glioblastoma second copy of the NF1 gene in a Schwann cell population multiforme. Brain 130, 2596-606. is an essential early step in the pathogenesis of both • Kunitz, A., et al. (2007). DNA hypermethylation and Aberrant neurofibroma and MPNST. The only well-characterized Expression of the EMP3 Gene at 19q13.3 in Human Gliomas. function of the NF1 gene product neurofibromin is its Brain Pathol. 17(4), 363-70. RasGAP activity, contained in the central GAP related • Weller, M., et al. (2007). Combined 1p/19q loss in oligodendroglial tumors: predictive or prognostic biomarker? Clin. Cancer domain. Neurofibromin function also seems to be involved Res. 13(23), 6933-7. in the cAMP protein kinase A (PKA-) pathway. We aim at • Balss, J., et al. (2008). Analysis of the IDH1 codon 132 mutation identification and characterisation of new Ras-dependent in brain tumors. Acta Neuropathol. 116. and Ras-independent functions of neurofibromin. • Kleber, S., et al. (2008). Yes and PI3K bind CD95 to signal inva- Furthermore we focus on the identification of potential sion of glioblastoma. Cancer Cell 13(3), 235-48. new therapeutic targets to target Schwann cells harbouring • Siegelin, M., et al. (2008). The flavonoid Kaempferol sensitizes a somatic NF1 mutation thus allowing a therapy that does human glioma cells to TRAIL-mediated apoptosis. Mol. Cancer not exhibit substantial toxic side effects on other cell types. Ther. 7(11), 3566-74. As new tumors arise throughout the life of patients with • Michaelis, M., et al. (2009). Reversal of P-glycoprotein-mediated multi-drug resistance by the MDM2 antagonist nutlin-3. NF1, we are also interested in evaluating mechanisms of Cancer Res. 69(2), 416-21. tumor immune escape in peripheral nerve sheath tumors and their implications for pathogenesis and therapeutic approaches.

Apoptosis in malignant gliomas (Dr. M. Siegelin) Despite aggressive multimodal treatment, malignant gliomas are almost fatal. The resistance to apoptosis contributes to low radiation and drug sensitivity of glioma cells. The presence of high levels of anti-apoptotic factors, such as the inhibitor of apoptosis proteins (IAP) confers resistance to cancer cells. One of our interests focuses on Structure of the Group IAP-Proteins and their antagonists. For this purpose, we analyze protein interactions between hypoxia-induced Group Leader: Andreas von Deimling proteins and IAP proteins. Special attention is given to Scientists: Christian Hartmann, Wolf Müller, migration, proliferation and apoptosis of hypoxic glioma David Reuss, Jörg Balß, David Capper, cells in respect to modifications in IAPs. Andrey Korshunov Technicians: Antje Habel, Ulrike Lass, Jochen Meier, Franziska Mößler, Jana Mucha, Kerstin Weber

53 Winfried Denk Top publications • Margrie, T. W., et al. (2003). Targeted whole-cell recordings in Structure of and the mammalian brain in vivo. Neuron 39, 911-918. activity in neuronal • Denk, W., et al. (2004). Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostruc- networks ture. Plos Biol. 2, 1900-1909. • Hasan, M. T., et al. (2004). Functional fluorescent Ca2+ indicator proteins in transgenic mice under TET control. Plos Biol. 2, 763-775. • Helmchen, F., et al. (2005). Deep tissue two-photon microscopy. Nat. Meth. 2, 932-940. • Briggman, K. L., et al. (2006). Towards neural circuit reconstruction with volume electron microscopy techniques. Curr. Opin. Neurobiol. 16, 562-570. Curriculum Vitae • Rueckel, M., et al. (2006). Adaptive wavefront correction in Degrees: 1984 Diploma in Physics two-photon microscopy using coherence-gated wavefront 1990 Ph.D. in Physics sensing. Proc. Natl. Acad. Sci. USA 103, 17137-17142. 1978-1981: Studies in Physics, • Hausselt, S. E., et al. (2007). A dendrite-autonomous mecha- Ludwigs-Maximilians Universität, Munich nism for direction selectivity in retinal starburst amacrine cells. 1981-1984: Studies in Physics, Eidgenössische Technische Plos Biol. 5, 1474-1493. Hochschule (ETH), Zurich, Switzerland • Hennig, P., et al. (2007). Point-spread functions for backscat- 1984-1989: Graduate Studies, tered imaging in the scanning electron microscope. J. Appl. Cornell University, Ithaca, NY, USA Physics. 102, 123101-01-123101-08. 1989: PhD in physics (1990) PhD thesis: • Kerr, J. N. D., et al. (2008). Imaging in vivo: watching the brain “Biophysical studies on saccular hair cells” in action. Nat. Rev. Neurosci. 9, 195-205. in the Laboratory of Prof. Watt W. Webb • Kuhn, B., et al. (2008). In vivo two-photon voltage-sensitive 1989-1991: Postdoctoral Research fellow, IBM Research Lab in dye imaging reveals top-down control of cortical layers 1 and Rueschlikon, Switzerland 2 during wakefulness. Proc. Natl. Acad. Sci. USA 105, 7588- 1991-1999: Independent Research fellow, Bell Laboratories, 7593. Murray Hill NJ Member of the Technical Staff, Biological Computation Research Department since 1999: Director of the department for Biomedical Optics at the Max Planck Institute for Medical Research, Heidelberg, Germany since 2002: Adjunct Professor, Faculty of Physics, University of Heidelberg Contact Department of Biomedical Optics MPI for Medical Research Jahnstraße 29, D-69120 Heidelberg A +49 (0) 6221 486 335 E +49 (0) 6221 486 325 K [email protected] o http://www.mpimf-heidelberg.mpg.de/abteilungen/ biomedizinischeOptik/denk/

54 Current Research Andreas Draguhn Functional analysis of cortical neuronal Function and plasticity of networks GABAergic synapses Research Summary The complex organisation Our group is interested in neuronal communication and of central synapses offers the resulting functional organization of neuronal net- multiple mechanisms for works. We focus on memory-related systems, especially regulation and modulation the mammalian hippocampus and neighbouring structu- of synaptic strength. We res. Using mostly electrophysiological methods, we ana- focus on inhibitory synapses lyse the function and plasticity of inhibitory synapses and in the mammalian CNS which fast network oscillations, trying to elucidate conditions use GABA (gamma-aminobutyric acid) as transmitter. In and mechanisms of information processing. previous work, we and others have provided evidence that changes in presynaptic GABA content do change the efficacy of inhibitory transmission. The availability of GABA Curriculum Vitae is regulated by its uptake, synthesis and degradation. The Degrees: 1991 MD University of Heidelberg relative contribution of these functions can change in 1999 Habilitation Humboldt University Berlin 1980-1987: Studies in medicine, physics and philosophy at situations of enhanced or reduced activity. Increased Bonn University uptake of GABA, for example, may occur under conditions 1987-1990: Thesis work (MPI for Biophysical Chemistry, of high activity when exocytosis of GABA from synaptic Göttingen, MPI for Medical Research, Heidelberg) terminals is enhanced. This will, in turn, increase recycling 1991: Residency (PJ) in Remscheid, Germany of the inhibitory transmitter and thus stabilize inhibitory 1992-1994: Research assistant, Dept. of Physiology, transmission. Prolonged periods of enhanced activity University of Cologne, Germany result in increased expression of the GABA-synthetizing 1994-1997: Research assistant, Dept. of Physiology, enzyme GAD while a chronic decrease of neuronal Humboldt University Berlin (Charité) activity leads to a down-regulation of GAD. Thus, multiple 1997: Senior research fellow, University of Birmingham Medical School, Birmingham, UK mechanisms seem to act together to adapt the efficacy of 1998-2002: Senior research assistant, Dept of Physiology, GABAergic transmission to the degree of activity in the local Humboldt University Berlin (Charité) network. These feedback loops constitute a mechanism since 2002: Professor of Physiology and Chair, Institute of of homeostatic network plasticity. We are presently Physiology and Pathophysiology, testing this hypothesis using various electrophysiological, University of Heidelberg, Germany histological and biochemical techniques. We also have established a model system for epileptiform discharges in Contact mouse brain slices in vitro. We use these pathophysiological Institute for Physiology and Pathophysiology patterns of network activity for the analysis of different University of Heidelberg pharmacological approaches which aim at increasing Im Neuenheimer Feld 326 presynaptic levels of GABA. In a third line of research, we 69120 Heidelberg Germany study GABAergic signalling in the developing nervous A +49-6221-544057 system, focussing on inhibitory interneurons in the early E +49-06221-546364 postnatal dentate gyrus. This work is integrated into the K [email protected] joined research cluster SFB488. o http://physiologie.uni-hd.de/draguhn/agr_dra.html

55 Andreas Draguhn

Leading questions: High-frequency network oscillations in the mouse hippocampus • How does the presynaptic concentration of GABA affect inhibitory synaptic transmission? Rhythmic activity is a key functional feature of the brain, • Which transport systems are involved in regulating as evident from the EEG. Meanwhile, many neuroscientists presynaptic GABA? agree that such network oscillations are “meaningful” and provide an important background for temporal coding • How is GABA release modulated by presynaptic of information. The basic concept is that synchronized ionotropic GABA receptors? oscillations bind neurons into transient assemblies where • How are tonic and phasic activation of GABA receptors co-active neurons constitute transient representations regulated during ontogenesis? (e.g., spatial memories). In our group we focus on one • Can we identify pharmacological means to enhance type of network oscilations in the rodent hippocampus, presynaptic GABA content in epilepsy? namely “ripples” at ~200 Hz as originally described by John O´Keefe, G. Buszáki and others. We are studying high- frequency oscillations in rodent hippocampal slices in vitro. In previous work we have described the crucial role of gap junctions for neuronal synchronization at this fast scale. Presently, we are trying to elucidate the functional properties of those cells which participate in ripples. We are also addressing pathophysiological questions regarding impaired memory in depressive patients. Here, we focus on alterations of high-frequency network oscillations by corticosteroids. This work is done in cooperation with several groups joined in the research cluster SFB636.

Leading questions: • Do members of transient neuronal assemblies have distinct electrophysiological properties? • How does the information encoded by transient assemblies propagate through the hippocampal loop? • How many functional assemblies do coexist in the hippocampus? Ho stable or plastic are they? • Are fast hippocampal network oscillations influenced by changing levels of glucocorticoids? Fig. 1: Plasticity of GABAergic synapses Top: GABA is synthesized from glutamate and packed into synaptic vesicles. Sources of GABA are GABA-uptake by GAT-1 and glutamate- Further projects: uptake by EAAC1. These membrane-bound transporters compete with Role of glutamate-uptake in epileptic seizures uptake of transmitters into adjacent glia cells. • Bottom: Increased strength of GABAergic transmission following high- (cooperation with Department of Pediatric Neurology); frequency stimulation of stratum radiatum in CA1. The right panel shows • Function of native and mutated inwardly rectifying enlarged miniature inhibitory postsynaptic currents following stimulation. potassium channels (cooperation with Clinics of This increase is due to the release of both, GABA and glutamate, which Cardiology); are subsequently taken up into the terminal and increase vesicular GABA content. Thereby, activity within the network is linked to the strength of • Pathogenetic mechanisms of Alzheimer´s disease inhibition. (cooperation with Abbott GmbH, Ludwigshafen).

56 Andreas Draguhn

Top publications • Draguhn, A., et al. (1998). Electrical coupling underlies high- frequency oscillations in the hippocampus in vitro. Nature 394, 189-192. • Traub, R.D., et al. (1999). High-frequency population oscillations are predicted to occur in hippocampal pyramidal neuronal networks interconnected by axoaxonal gap junctions. Neurosci. 92, 407-426. • Engel, D., et al. (2001). Plasticity of rat central inhibitory synapses through GABA metabolism. J. Physiol. 535, 473-482. • Maier, N., et al. (2003). Cellular and network mechanisms underlying spontaneous sharp wave-ripple complexes in mouse hippocampal slices. J. Physiol. 550, 873-887. • Buzsaki, G., et al. (2004). Neuronal oscillations in cortical networks. Science 304, 1926-1929. • Nimmrich, V., et al. (2005). Induced sharp wave-ripple complexes in the absence of synaptic inhibition in mouse hippocampal slices. J. Physiol. 563, 663-670. • Both, M., et al. (2008). Propagation of specific network patterns through the mouse hippocampus. Hippocampus 18, 899-908. • Hartmann, K., et al. (2008). Fast homeostatic plasticity of inhibition via activity-dependent vesicular filling. PLOS One 3, e2979. • Nimmrich, V., et al. (2008). Amyloid β oligomers (Aβ1-42 Glo- Fig. 2: Analysis of fast network oscillations bulomer) suppress spontaneous synaptic activity by inhibiti- Top: Schematic drawing of a neuronal assembly in CA1. The highlighted on of P/Q-type calcium currents. J. Neurosci. 28, 788-97. neurons are selectively activated during a network event (e.g., a high- frequency ripple oscillation) and elicit action potentials in a fixed temporal sequence. All other cells are suppressed by synaptic inhibition. Bottom: Field potential recordings of sharp wave-ripple complexes from a mouse hippocampal slice. Right panel shows extended view of one spontaneous network event. Bottom traces show isolated unit discharges, filtered field potential with oscillation at ripple frequency and original recording. Right: Cross-correlogram between units and field ripples showing the tight synchronization of single action potentials by the underlying network oscillation. Structure of the Group Group Leader: Andreas Draguhn Postdoctoral fellows: Martin Both, Jurij Brackack, Claus Bruehl, Valeri Lopantsev, Astrid Bertsche (guest) PhD-Students: Florian Bähner, Gunnar Birke, Nana Duhme, Martin Oehmen, Susanne Reichinnek, Jan Schönberger, Elisa Weiss, Maura Zylla Undergraduates: Nils Kasties, Beate Hienz Technicians: Andrea Lewen, Nadine Zuber Secretaries: Elke Jochum, Ute Schmitt

57 Uwe Ernsberger Current Research

Development of nerve cells and generation of In recent years, significant neuronal diversity progress has been made to understand the generation Research Summary of neuronal diversity. One of the key models used for To understand how population-specific and general the study of this question is neuronal properties are acquired during neuronal the peripheral sympathetic development, we analyze the expression of genes coding nervous system of mammals for neurotransmitter-synthesizing enzymes and synaptic and birds. Sympathetic proteins. In sympathetic ganglia growth factors from the ganglia contain two neuron BMP family induce early widespread expression of both populations which develop from the same precursor classes of genes. GDNF family ligands are later involved in pool but differ in their transmitter, noradrenaline or the differentiation of neuronal subtypes. acetylcholine, respectively. Analyzing the expression of Curriculum Vitae genes coding for the transmitter-synthesizing enzymes, our interest is to identify growth factors and transcription Degrees: 1985 M.Sc. factors which are crucially involved in the segregation of 1988 Ph.D. neuronal fates. In addition, we analyze signals regulating 2000 Habilitation 1985-1988: Thesis work (MPI for Psychiatry, Martinsried the expression of general neuronal genes such as Christian-Albrechts-Universität, Kiel) those coding for synaptic proteins to understand how 1988-1991: Postdoctoral fellow, general neuronal differentiation and population-specific Department for Biology , development are coordinated. University of California, San Diego, USA 1992-1998: Postdoctoral fellow in the Department for Noradrenergic induction - specifying neurons by Neurochemistry, MPI for Brain Research, Frankfurt coordinate gene expression 1999-2004: Postdoctoral fellow, in the Department for Anatomy and Cell Biology III, Noradrenergic properties are induced early during University of Heidelberg sympathetic neurogenesis by BMP growth factors and since 2000: Group Leader, Neuroanatomy, University of Heidel- Phox2 transcription factors in birds and mammals. berg Comparing the expression of tyrosine hydroxylase and dopamine β-hydroxylase, we could show that these two Contact enzymes of the noradrenaline biosynthesis cascade are Department of Neuroanatomy expressed at the same time during sympathetic neuron Institute for Anatomy and Cell Biology differentiation. The induction by the same growth and Interdisciplinary Center for Neurosciences transcription factors suggests regulation of these enzymes Im Neuenheimer Feld 307 as a synexpression group that is conserved in evolution 69120 Heidelberg Germany from birds to mammals. A +49-6221-548344 E +49-6221-545604 Genes coding for synaptic proteins - induction of general K [email protected] as compared to population-specific properties o http://izn.uni-hd.de/ Synaptic proteins are more generally expressed in different classes of neurons than individual transmitter-synthesizing

58 Uwe Ernsberger

genes provides evidence for early segregation of neuronal and endocrine lineages.

Generation of neuronal diversity in sympathetic ganglia Choline acetyltransferase (ChAT) and the vesicular acetylcholine transporter (VAChT) are two main features of cholinergic neurons which are coordinately transcribed from the cholinergic gene locus. The loss of ChAT and VAChT expression in sympathetic ganglia of c-ret mutant Fig. 1: Schematic representation of a transmitter vesicle, the transmitter mice demonstrates that signaling via receptors for growth synthesizing enzymes tyrosine hydroxylase (TH) and dopamine factors of the GDNF family is necessary for the development b-hydroxylase (DBH), and certain proteins involved in vesicle fusion and transmitter release. SNAP-25 (S25), synaptotagmins (SYT), neurexins of cholinergic sympathetic neurons. Expression of c-ret (NEU), and voltage-gated calcium channels (Ca-K) are involved in in cholinergic sympathetic neurons of the chick embryo different functional aspects of transmitter release and are addressed in suggests that its role may be evolutionary conserved. our studies. Whereas c-ret acts late during maturation in sympathetic ganglia, factors involved in the early induction of enzymes which are restricted to certain neuron populations. cholinergic properties remain to be determined. They function in different aspects of transmitter release and synapse organization. Overexpression studies show that synaptotagmin I and neurexin I can be induced by BMP growth factors and Phox2 transcription factors in sympathetic precursors. Thus, it appears that induction of general and population-specific neuronal characters involves common mechanisms. In the closely related adrenal chromaffin cells, different regulation of these

Fig. 3: Expression of mRNAs for tyrosine hydroxylase (TH) [red] and Fig. 2: Detecion of mRNAs for dopamine β-hydroxylase (DBH) (A) and neurofilament M (NF-M) [black] in sympathoadrenal precursors. Distinct ret (B) by in situ hybridization shows that the vast majority of neurons expression levels for the subclass-specific marker TH and the general in the superior cervical ganglion are noradrenergic. A subpopulation of neuronal marker NF-M demonstrates different regulation of gene neurons expresses ret. expression in neuronal and neuroendocrine cells.

59 Uwe Ernsberger

Future Research and Goals Top publications Our interest is to understand how signalling by • Burau, K., et al. (2004). C-ret regulates cholinergic properties in growth factors and regulation by transcription factors mouse sympathetic neurons: evidence from mutant mice. Eur. coordinates the acquisition of general neuronal features J. Neurosci. 20, 353-362. and the diversification of neuronal lineages. Analyzing • Ernsberger, U. (2004). Gene expression from precursor to mature neuron. In Molecular Biology of the Neuron, 2nd ed; W. developmental expression patterns of genes coding for Davies, et al., eds. (Oxford, UK: Oxford University Press), pp different synaptic proteins which are expressed across 29-73. many neuron populations, we ask whether different • Ernsberger, U., et al. (2005). Expression of neuronal markers neuron classes share common differentiation pathways. suggests heterogeneity of chick sympathoadrenal cells prior We then search for the transcriptional regulators involved to invasion of the adrenal anlagen. Cell Tissue Res. 319, 1-13. to understand whether general neuronal features are • Ernsberger, U., et al. (2006). Phox2a and Phox2b: essential induced by population-specific or rather by widely transcription factors for neuron specification and differentiati- expressed transcription factors. In comparison, we on. In Transcription factors in the nervous system, G. Thiel, ed. investigate the expression of genes coding for enzymes (Weinheim, Germany: Wiley-Verlag Chemie), pp 53-74. • Huber, K., et al. (2006). Cholinergic differentiation occurs ear- of neurotransmitter biosynthesis and transport which are ly in mouse sympathetic neurons and requires Phox2b. Gene specific for defined neuron populations. Understanding Expr. 13, 133-139. the factors involved in their expression, we hope to • Ernsberger, U. (2008). The role of GDNF family ligand signal- comprehend how general and population-specific aspects ling in the differentiation of sympathetic and dorsal root gan- of neuronal differentiation are coordinated. glion neurons. Cell Tissue Res. 333, 353-371.

60 Current Research Thomas Euler Signal processing in the retina Visual information processing begins in the retina. Sti- Research Summary mulus features, such as spatial extent, intensity, color, Our group studies the processing of visual information edges, motion, direction etc., in the retina using electro- and multi-photon opto- are extracted for each retinal physiological techniques. We focus on the question how location, involving circuits dendrites process information. This is of particular interest built from various types in the retina, where the largest class of interneurons, the (~70) of neurons. This infor- amacrine cells, typically lack axons and use their dendrites mation is coded for transmis- both for receiving input and making output synapses. sion via the optic nerve to higher visual centers. The retina is a part of the brain that can be easily isolated and stays Curriculum Vitae fully functional for hours. The tissue is highly transparent Degrees: 1993 Diploma, University of Mainz and, thus, ideal for optical recordings using multi-photon 1996 Dr. rer. nat. (Biology), University of Mainz microscopy (Denk & Detwiler, 1999, PNAS 96:7035-40). Fur- 1993-1996: Thesis work (University of Mainz / thermore, the retina is amenable to genetic manipulations, MPI for Brain Research, Frankfurt) 1996-1997: Postdoctoral fellow for example by viral gene transfer via intraocular injection, (MPI for Brain Research, Frankfurt) which, unlike for other part or the brain, does not require 1997-2000: Postdoctoral fellow (Harvard Medical School / surgery and stereotactical targeting. Mass. General Hospital, Boston, USA) since 2000: in the Dept. of Biomed. Optics, Computation of image motion direction in the retina MPI for Medical Research, Heidelberg 2000-2003: Research fellow One type of interneuron we study (in close collaboration 2003-2006: Project/Group Leader with P. Detwiler, University of Washington, Seattle, and W. since 2006: Research Group Leader Denk) is the so-called ‘starburst’ amacrine cell (SAC, Fig. 1). since 2004: Teaching certificate (University of Heidelberg) SACs are presynaptic to direction-selective (DS) ganglion cells, which fire strongly when a visual stimulus moves in Contact a certain direction but remain silent when the stimulus Department of Biomedical Optics moves into the opposite direction (Barlow et al., 1964, J MPI for Medical Research Physiol 173:377-407). Earlier studies have shown that DS Jahnstr. 29 ganglion cells receive directionally tuned synaptic input 69120 Heidelberg (reviewed in Demb, 2007, Neuron 55:179-186). With multi- Germany photon Ca2+ imaging of SAC dendrites we have shown A +49-6221-486320 E that light stimuli moving from the soma to the dendritic +49-06221-486325 K [email protected] tips (centrifugal, CF) elicit larger Ca2+ signals than motion o http://www.mpimf-heidelberg.mpg.de/~teuler/ in the opposite direction (centripetal, CP). Thus, SAC dendrites provide a directionally tuned output signal. The SAC’s dendritic sectors operate largely independently and can be viewed as processing units that signal CF motion. We studied the mechanisms underlying the computation of DS signals in SAC dendrites by measuring the electrical

61 Thomas Euler responses to CF and CP moving stimuli using whole-cell relationship between non-linearity and holding potential patch-clamp recordings. Spectral analysis revealed larger indicates motion direction-dependent activation of non-linearities in the electrical response to CF motion voltage-gated channels, pointing at a dendritic DS than to CP motion. Dendritic DS persists in the absence mechanism that relies on intrinsic properties. of inhibitory transmission, which indicates that inhibitory Calcium changes in SAC dendrites evoked by voltage steps network interactions are not essential for DS in SACs. The and monitored by multi-photon imaging (Fig. 1) indicate that Ca2+ channels are active at rest. The data suggests that, in part due to glutamatergic input, the distal dendrite is tonically depolarized relative to the soma. The resulting somato-dendritic voltage gradient is likely a key element in “dendrite-autonomous” DS computation, because it allows distal Ca2+ channels to operate at a point in their activation range where the gain is maximal and small voltage deflections result in large currents. In related experiments we map light-stimulus evoked Ca2+ signals along SAC dendrites and apply pharmacology. Here we aim to determine the contribution of different types of Ca2+ channels to the SAC response. Further we want to find out if intracellular Ca2+ signaling, which is known to play a role in transmitter release from amacrine cells, contributes to the SAC´s output signals.

Bipolar cells are parallel information channels in the retina Another focus of the group is on signal processing in retinal bipolar cells. For example, we studied the role of intracellular [Cl-] gradients and their dynamics in neuronal processing (in collaboration with T. Kuner). ON bipolar cells, which are depolarized by light, have long been proposed Fig. 1: Starburst amacrine cell (a) in the rabbit retina filled with fluorescent - Ca2+ indicator via a patch-clamp electrode (from left) in voltage-clamp to sustain an axo-dendritic [Cl ] gradient that allows these configuration. Right: [Ca2+]-dependent fluorescence (in arbitrary units), cells to process GABAergic input differentially at their two recorded by multi-photon microscopy, increased in a distal dendritic ends, the dendrites and the axon terminal. branch when stepping the somatic holding voltage (VCOM) from -75 To test this hypothesis we used ratiometric multi-photon (upper panel) to -2 mV (lower panel; averages of 15 frames). Changes - in somatic current (Im) and dendritic [Ca2+] (b) evoked by somatic microscopy and mapped the local [Cl ] in bipolar cells voltage steps (different cell than in a). The hyperpolarizing step induces expressing the genetically-encoded Cll- indicator a decrease in [Ca2+], indicating that voltage-gated Ca2+ channels in the Clomeleon (Kuner & Augustine, 2000, Neuron 27, 447-459). distal dendrites are open at the somatic resting potential. SACs contain - 2+ We found that ON bipolar cells generate [Cl ] gradients only high-voltage activated (HVA) Ca channels, therefore the dendrites - must be tonically depolarized relative to the soma to allow HVA channels with high [Cl ] in their dendrites (Fig. 2), allowing these to be activated. Ca2+ responses as a function of VCOM (c) before (black) cells to receive via the same neurotransmitter, GABA, during (red) and after (blue) bath application of CNQX (n=3 cells; lines: excitation at their dendrites and inhibition at their axon fitted activation curves). Blocking CNQX-sensitive input shifts the terminal. Dendritic [Cl-] is particularly high in so-called [Ca2+] vs. voltage curve towards depolarized potentials, indicating that a tonic glutamatergic inward current is, at least in part, responsible for blue-cone bipolar cells, which receive input exclusively depolarizing the dendrites. (b: traces are single trials) from short-wavelength (‘blue’) sensitive cones. To find

62 Thomas Euler out if high dendritic [Cl-] in these bipolar cells is playing a Top publications particular role in chromatic processing, we now study the • Euler, T. (2003). Richtungsmelder im Auge. Gehirn & Geist (Ver- blue-cone circuit in mouse retina with both anatomical (in lag Spektrum der Wissenschaft) 4, 64-67. collaboration with Silke Haverkamp, MPIH, Frankfurt) and • Huang, L., et al. (2003). The novel G protein subunit Ggam- physiological techniques. ma(13) is co-expressed with Galpha(o), Gbeta(3) and Gbeta(4) in retinal ON bipolar cells. J. Comp. Neurol. 455, 1-10. • Hasan, M.T., et al. (2004). Functional fluorescent Ca2+ indicator proteins in transgenic mice under TET control. PLoS Biol. 2(6), June, 15. • Haverkamp, S., et al. (2005). The primordial, blue cone color system of the mouse retina. J. Neurosci. 25, 5438-5445. • Oesch N, et al. (2005). Direction-selective dendritic action potentials in rabbit retina. Neuron 47, 739-750. • Duebel J., et al. (2006). Two-photon imaging reveals somato- dendritic chloride gradient in retinal ON-type bipolar cells expressing the biosensor Clomeleon. Neuron 49, 91-94. • Hausselt, S.E., et al. (2007). Dendrite-autonomous computation of motion direction in starburst amacrine cells. PLoS Biolo- gy 5(7), e185. • Euler, T., et al. (2008). Eyecup Scope – Optical recordings of light stimulus-evoked fluorescence signals in the retina. Pflü- gers Arch./Europ. J. Physiol., DOI 10.1007/s00424-008-0603-5. • Margolis, D.J., et al. (2008). Functional Stability of Retinal Gan- glion Cells after Degeneration-Induced Changes in Synaptic Input. J. Neurosci. 28, 6526-36. Fig. 2: Clomeleon-expressing bipolar cells (A) injected with fluorescent • Euler, T. et al. (2008). Direction Selective Cells. In The Senses – dye (red) and imaged with multi-photon microscopy (fixed section; A Comprehensive Reference, Vol. 1 Vision I, A.I. Basbaum, et al., Clomeleon labeling in green). Several retinal layers can be distinguished: eds., (San Diego, USA: Academic Press). 413-422. OPL: outer plexiform layer; INL: inner nuclear layer; IPL: inner plexiform layer; double-labeled bipolar cell in yellow. Fixed flat-mounted retina (B) co-labeled with antibodies against GluR5 (red) revealing cone pedicles. A subset of Clomeleon bipolar cells (*) contacts only few pedicles (examples encircled), which belong to blue-sensitive cones [5]. Blue cone-selective bipolar cells (C) marked by asterisks. Left: YFP fluorescence; right: map of intracellular Cl- concentration ([Cl-]). [Cl-] is high in the varicosities (arrowheads), where cone pedicles are contacted. Traces showing dendritic (red) and somatic (blue) [Cl-] (D) in a blue cone-selective bipolar cell during GABA application and during co-application of GABA antagonists. The transient decrease in dendritic [Cl-] suggests that GABA mediates depolarization in blue cone-selective bipolar cells. (Scale bars: 20µ ; a-c: collapsed image stacks; d: traces are averages of 3 trials)

Structure of the Group Group Leader: Thomas Euler Postdoctoral fellows: Xavier Castell, Julia Mack-Bucher PhD-Students: Tobias Breuninger, Minggang Chen, Sun Le

63 Christian Fiebach Current Research

Cognitive Neuroscience: Neural mechanisms Our research aims at of higher cognitive functions understanding the neural mechanisms that underlie Research Summary higher cognitive functions in humans, such as We use modern, non-invasive neuroimaging techniques language, working memory, such as functional magnetic resonance imaging and elec- or goal-directed behavior tro- and magnetoencephalography to explore the neural in general. To this end, we mechanisms underlying higher cognitive functions in employ functional magnetic humans. Our main areas of research involve cognitive resonance imaging to functions such as language, working memory, decision identify brain regions that are specifically activated under making, and goal directed behavior. In addition, we are certain, highly controlled cognitive demands. In addition, particularly interested in specifying the neural mecha- we also explore how different brain regions are functionally nisms underlying individual differences in these cogniti- coupled during specific task demands, using statistical ve functions. methods that exploit the functional connectivities Curriculum Vitae between brain regions. Degrees: 1998 Diploma ( M. Sc.) Neural bases of individual differences 2001 Ph.D. (Dr. rer. nat. Psychology) 1998-2001: Thesis work, MPI for Human Cognitive and Neural mechanisms underlying inter-individual differences Brain Sciences, Leipzig in working memory capacity and higher cognitive abilities 2001-2003: Postdoctoral fellow, MPI for Human Cognitive (intelligence, complex problem solving) are poorly and Brain Sciences, Leipzig understood. Three potential sources of variability may 2003-2006: Postdoctoral fellow, Department of Psychology contribute to these higher cognitive abilities: variability & Helen Wills Neuroscience Institute, University of California, Berkeley, USA in component processes of cognition, personality since 2006: Group Leader, Emmy-Noether Research group: differences, and variability of dopamine neurotransmitter “Neurocognition of individual differences“, activity. This research project integrates these at present Departments of Psychology, Neuroradiology and mostly unconnected research strands, and explores their Neurology, University of Heidelberg relations and their contribution to higher cognition at a neurocognitive level. Contact Department of Psychology Cortical representations of behavioral goals University of Heidelberg The ability to plan and solve complex problems is crucial Hauptstr. 47-51 69117 Heidelberg for intelligent human behavior in a multitude of domains Germany such as school, work, and everyday life. Planning and A +49-6221-547371 problem solving are driven by goals: We have problems K [email protected]’ to solve when our goals are not immediately achievable. o http://fimlab.uni-hd.de Such complex behaviors therefore require the ability to maintain goals and subgoals over time, to resolve goal conflicts, and to adapt behavioral goals to environmental changes. Human functional neuroimaging studies

64 Christian Fiebach

show that frontopolar cortex is strongly activated when memory maintenance (e.g., Fiebach et al., 2006; cf. Figure planning demands are high. This may be a result of 1). The aim of this work is to model the neurocognitive the need to perform one or more subgoal tasks in the bases of verbal working memory using domain-general, service of a primary task goal: Subgoal processing, or neurophysiologically plausible organizational principles. ‘cognitive branching’ selectively activates the frontopolar cortex. It is unlikely that frontopolar cortex processes Brain mechanisms of emotion x cognition interaction goals and subgoals in isolation; rather, it is assumed that Traditionally, cognitive psychologists and cognitive frontopolar cortex functionally interacts with prefrontal neuroscientists investigate mental processes in isolation. and other task-relevant brain regions. In this project, However, from our everyday experience, we know that we use functional magnetic resonance imaging (fMRI) emotional states and arousal can greatly influence our and electroencephalography (EEG) to investigate the actual performance. For example, in a recent experiment, neurocognitive organization of various aspects of goal we demonstrate that performance in a working memory management and goal maintenance. task is modulated by the incidental presence of positive and negative emotion in the stimulus materials (Fig. 2). Neurocognition of verbal working memory We assume that the influence of emotion on cognition is Verbal working memory is often characterized in the mediated by functional interactions of neuroanatomical context of the multicomponent model of Baddeley and systems involved in these processing domains. We use collaborators. In this model, maintenance of linguistic fMRI and multivariate analyses of functional connectivities information in verbal working is strictly based on to explore how cognition and emotion interact in working phonological codes. This postulate of a special purpose memory and decision making. mechanism for verbal working memory is inconsistent with working memory models based on neurophysiological data, such as Fuster’s active memory model. In work started at the D’Esposito Neuroimaging Laboratory at UC Berkeley, we use fMRI to investigate the contribution of different language systems (i.e., visual-orthographical, lexical, semantic, phonological) to verbal working

Fig. 2: Emotion by cognition interaction in a verbal working memory task. Percentage of correct responses dependent on working memory load (2 words vs. 5 words) and valence (red: negative; grey: neutral; green: positive). A valence-by-cognition interaction shows that under low cognitive demands (i.e., low working memory load), subjects perform Fig. 1: (left panel) Word-sensitive region in left inferotemporal cortex. better for positive and negative relative to neutral items. Under high (right panel) Working memory modulation of brain activation of the cognitive demand, however, only negative emotion has a beneficial inferotemporal region displayed in (left panel), dependent on stimulus influence on performance. This suggests the existence of mechanisms type (words vs. pseudowords) and working memory load (green: 2 items; that prioritize potentially threatening information, even under high orange: 5 items) cognitive resource usage.

65 Christian Fiebach

Functional neuroanatomy of the language system Top publications The ability to produce and understand language is one of • Fiebach, C. J., et al. (2003). Processing concrete words: fMRI the greatest cultural achievements of mankind. Using tech- evidence against a specific right hemispheric involvement. niques such as functional magnetic resonance imaging, Neuropsychologia, 42, 62-70. electroencephalography and magnetoencephalography, • Fiebach, C. J., et al. (2003). Distinct brain representations for early and late learned words. NeuroImage,19, 1627-1637. we study neural mechanisms underlying the recognition • Fiebach, C. J., et al. (2004). Revisiting the localization of syn- of written and spoken words, as well as the mechanisms tax: Syntactic integration vs. syntactic working memory. Hum. underlying the comprehension of complex syntactic struc- Brain Mapp., 24, 79-91. tures (i.e., complex sentences). In Figure 3, data is presen- • Fiebach, C. J., et al. (2005). Neuronal mechanisms of repetiti- ted that demonstrates the involvement of prefrontal cor- on priming in occipito-temporal cortex: Spatio-temporal evi- tex in interference resolution during word recognition, in dence from fMRI and EEG. J. Neurosci., 25, 3414-3422. cases where the orthographic neighborhood of a letter • Fiebach, C. J., et al. (2006). Modulation of inferotemporal cor- string stimulus conflicts with its lexical status (Fiebach et tex during verbal working memory maintenance. Neuron, 51, al., 2007). 251-261. • Fiebach, C. J., et al. (2007). Infero-temporal cortex maintains conceptual-semantic representations in verbal working memory. J. Cogn. Neurosci., 19, 2035-2049. • Fiebach, C. J., et al. (2007). Inhibition and facilitation in visual word recognition: Prefrontal contribution to the orthographic neighborhood size effect. NeuroImage, 36, 901-911. • Stelzel, C., et al. (2009). Effects of dopamine-related gene- gene interactions on working memory component processes. EJN. 29, 1056-1063

Fig. 3: Left dorsolateral prefrontal cortex area modulated by lexicality (words vs. nonwords) and orthographic neighborhood size (N; small vs. large). Behaviorally, it has been shown that a large orthographic neigh- Structure of the Group borhood, i.e., the presence of many words that are similar to the target Group Leader: Christian Fiebach nonword, renders lexical decisions difficult. In the present fMRI study, we demonstrate that the left dorsolateral prefrontal cortex is particularly Postdoctoral fellows: Christine Stelzel, Alexander Roth strongly activated for nonwords with a large neighborhood, presumably PhD Students: Ulrike Basten, Andrea Gäbel, in an effort to resolve the conflict between target nonword and global Sascha Purmann, Ruth Schmitt lexical activation induced by the target’s orthographical neighbors. Undergraduates: Diana Armbruster

66 Current Research Herta Flor Learning and neuronal plasticity Learning, neuronal plasticity and Research Summary psychopathology Our research focuses on the interaction of brain and be- Research on this topic tar- havior, in particular the question how behavior and expe- gets the development and rience influence neural processes and how neural proces- maintenance of emotional ses alter behavior and experience. A special focus is on memories and their neuro- the role of implicit learning and memory processes in the nal correlates in the context development and maintenance of mental and psycho- of psychopathology. Herta physiological disorders. This includes research on memo- Flor is the spokesperson of DFG collaborative research ry processes in chronic pain, tinnitus, anxiety disorders, grant 636 on „Learning, memory and neuronal plasticity: addiction and depression. The methods that are emplo- Implications for psychopathology“. Rather than taking a yed range from experimental psychology to noninvasive nosological approach to mental disorder, which leads to brain imaging and peripheral psychophysiology. the problem of overlapping psychopathological characteristics, similarities and differences of the underlying me- Curriculum Vitae chanisms of learning and neural plasticity across a variety Degrees: 1981 Dipl.-Psych. University of Tübingen of disorders are examined. The Flor group pursues the role 1984 Ph.D. University of Tübingen of basic associative and non-associative learning mecha- 1983-1990: Pre- and postdoctoral fellow and assistant nisms and their neuronal correlates, which are fundamen- professor, Yale University, Universities of Pittsburgh, Bonn, Tübingen tal for the understanding of disorders characterized by 1990-1991: Visiting Professor University of Marburg hyperexcitable neuronal fear circuits (e.g., post-traumatic 1991-1993: Heisenberg fellow stress disorder, social phobia), disorders characterized by 1993-1995: Professor of Somatic psychology, a deficit in associative learning of aversive consequences Humboldt University Berlin (e.g. psychopathy) as well as disorders characterized by al- 1995-2000: Full Professor of Clinical psychology and tered appetitive and aversive motivational processes (e.g. behavioral neuroscience, Humboldt depression, bipolar disorder, addiction). One major the- University Berlin me is the relative contribution of learning processes and since 2000: Full Professor and Chair, Institute of genetic variables in the development of these disorders. Neuropsychology and Clinical Psychology, Central Institute of Mental Health and Behavioral and combined pharmacological-behavioral University of Heidelberg interventions, that address learning related maladaptive plasticity are developed and their influence on cortical Contact and subcortical plasticity are studied. Department of Cognitive and Clinical Neuroscience The group is also involved in a collaborative EU grant that Central Institute of Mental Health studies reinforcement-related learning processes, their J 5 neuronal correlates and genetic determinants with respect 68159 Mannheim, Germany to the development of mental disorders in adolescents. A +49-621-17036302 Finally, aging-related alterations in memory and learning E +49-621-17036305 and associated neuronal plasticity are also investigated. K [email protected] o http://www.zi-mannheim.de

67 Herta Flor

phantom perception and phantom pain. The differential contribution of frontal and parietal brain areas to painful versus nonpainful phantoms and related body illusions is examined. Novel brain-based interventions that use brain-computer interfaces, transcranial magnetic stimulation, mirror treatment or virtual reality are examined in their capability to alter central processes involved in pain and abnormal somatosensory phenomena.

Fig. 1: T-maps revealing increases in functional coupling for the contrasts between genotype-groups during the early acquisition phase/extinction phase and genotype-dependent functional coupling during early acquisition between left amygdala and prefrontal cortex, and during the extinction phase between left prefrontal cortex and left amygdala.

Learning and neuronal plasticity in chronic pain and abnormal body sensation The Flor group was the first to describe the role of cortical reorganization for the development and maintenance of phantom limb pain as well as other pain syndromes such as chronic back pain and fibromyalgia. Within the Fig. 2: The mirror box experiment. When a mirror is placed between the framework of the DFG Clinical Research Group “Learning, two hands of healthy persons or between the intact and phantom hand in amputees, movements of the hand that can be seen in the mirror are Plasticity and Pain” (CRG 107) and within two BMBF-funded perceived as movements of the hand that is hidden behind the mirror (or collaborative grants „Pain Perception“ and „Neuropathic the phantom in amputees). The brain activation that is then seen in pri- Pain“ cortical and subcortical alterations related to chronic mary somatosensory and motor cortex is not only present contralateral pain are examined and novel treatment options that target to the moved hand but can also be seen in the hemisphere contralateral to the hand that is perceived in the mirror. Activation in the brain is thus the extinction of central pain memories by behavioral and also related to perceived not only to actual movement. pharmacological interventions are developed and tested. Another focus is the development of indicators for Neuronal plasticity in the auditory system differential assignments of treatments to psychologically and neurobiologically characterized subgroups of pain One important line of research concerns the contribution patients. In this context we are also studying how early pain of neuronal plasticity in the auditory system to the experience may affect the processing of nociceptive input development and maintenance of Tinnitus symptoms. in children and how this may induce neuronal plasticity After having demonstrated that Tinnitus phenomena of the somatosensory system. A related EU grant focuses are associated with a reorganization of the tonotopic upon the development of innovative procedures to assess map in primary auditory cortex and general cortical phantom phenomena and the exploration of the bases of hyperexcitability, we have developed several new

68 Herta Flor approaches to the treatment of Tinnitus. These include Top publications behavioral auditory discrimination and extinction trainings • Birbaumer, N.,et al. (2005). Deficient fear conditioning in psy- with or without additional pharmacological interventions chopathy: A functional magnetic resonance imaging study. designed to enhance extinction processes. Our most Arch. Gen. Psych. 62, 799-805. recent research focuses on the interaction of limbic and • Heinz, A., et al. (2005). Amygdala–prefrontal coupling de- auditory areas in the processing of emotional stimuli in pends on a genetic variation of the serotonin transporter. Nat. Tinnitus patients and factors that contribute to the long- Neurosci. 8, 20-21. term-development of Tinnitus. This research is currently • Flor, H., et al. (2006). Phantom limb pain - a case of maladap- being extended to focus on affective-sensory interaction tive central nervous system plasticity? Nat. Rev. Neurosci. 7, and the brain processes involved in these interactions in 873-881. • Wessa, M., et al. (2006). Altered cortisol awakening response general. in posttraumatic stress disorder. Psychoneuroendocrinol. 31, 209-215. • Christmann, C., et al. (2007). A simultaneous EEG-fMRI study of painful electric stimulation. NeuroImage 34, 1428-1437. • Heinz, A., et al. (2007). Serotonin transporter genotype (5-HT- TLPR) effects of neutral and undefined conditions on amygdala activation. Biol. Psych. 61, 1011-1014. • Smolka, M. N., et al. (2007). Gene-gene effects on central processing of aversive stimuli. Mol. Psych., 12, 307-317. • Wessa, M. et al. (2007). Failure of extinction of fear responses in posttraumatic stress disorder: evidence from second-order conditioning. Am. J. Psych. 164, 1684-1692. Fig. 3: Tinnitus sufferers showed a higher BOLD response in the left • Hermann, C., et al. (2008). Cortical correlates of an attentional auditory association cortex (BA 22) to negative versus neutral pictures bias to painful and innocuous somatic stimuli in children with compared to controls. recurrent abdominal pain. Pain, [Epub ahead of print]. • Wölfling, K., et al. (2008). Psychophysiological responses to Method development drug-associated stimuli in chronic heavy cannabis use. EJN 27, 976-983. In all research areas methods of experimental psychology are combined with noninvasive brain imaging, brain stimulation and peripheral psychophysiological Structure of the Group methods as well as genetic testing and behavioral and Group Leader: Herta Flor pharmacological interventions to comprehensively study Postdoctoral fellows: Beate Herbert, Christiane Hermann, brain-behavior interactions in healthy humans and patients. Eugen Diesch, Carsten Diener, Martin Diers, A special focus is on the development of multimodal Alexander Kroll, Anita Kult, Simone Lang, imaging methods that integrate structural and functional Christoph Oberthuer, Kati Thieme, aspects and trace also dynamic alterations of cortical and Michèle Wessa subcortical changes related to the perceptual phenomena PhD-Students: Isabelle Bomba, Wenke Brusniak, studied by the group. This also includes the analyses of the Jens Foell, Johanna Hohmeister, Sandra interaction between peripheral and central physiological Kamping, Claudia Liebscher, Slawomira Lipinski, Maurice Moayer, Julia Ofer, changes. These methodological developments are made in Stephanie Ridder, Iris Wollgarten, close interaction with the members of the South German Pinar Yilmaz, Katrin Zohsel Brain Imaging Center that includes researchers from the Technicians: Annette Hornbach, Michael Rehm, Birgül Universities of Mannheim and Heidelberg as well as the Sarun, Claudia Stief, Heike Schmidt, Central Institute of Mental Health. Artemis Tsoupas, Angelika Wieters

69 Stephan Frings Current Research Signal transduction in sensory neurons Regulation of cAMP-gated transduction channels in Research Summary olfactory sensory neurons We examine the molecular pathways of signal transduction Upon detection of odorants in primary afferent neurons of the mammalian olfactory in the inhaled air, a and nociceptive systems. We concentrate on the transduction cascade is regulation of transduction channels and the generation initiated in the sensory cilia of excitatory receptor potentials. Of particular interest of olfactory sensory neurons. for us is the question how these neurons adjust their cAMP is synthesized and sensitivity and bring about adaptation or sensitization. opens calcium-permeable transduction channels. These channels are gated by direct Curriculum Vitae binding of cAMP, and they are regulated by calcium/ Degrees: 1985 Diploma in Biology calmodulin. Activation and inhibition are mediated by 1989 Ph.D. modulatory channel subunits which determine cAMP- 1997 Habilitation in Zoology, sensitivity and calmodulin binding. We explore the allosteric University of Cologne regulatory mechanisms that operate in this channel. We 1980-1985: Studies of Biology at the University of Konstanz perform structure-function studies on heterologously 1985-1989: PhD Student at the University of expressed channels carrying mutations in key domains of Otago Medical School, Dunedin, New Zealand the heteromeric channel protein. 1989-1992: Postdoctoral fellow at the Department of Physiology, University of Saarland, Homburg/Saar We are particularly interested in the formation of a 1992-2002: Postdoctoral fellow at the Institute of Biological regulatory complex that consists of two cytoplasmic Information Processing, Jülich Research Center channel domains and calmodulin. We explore the since 2002: Professor for Molecular Physiology at Heidelberg stoichiometry of this complex in vitro by studying the University assembly of calmodulin with peptides representing the relevant channel domains. Our goal is to understand the Contact regulation of the cAMP-gated channels by calcium. Department of Molecular Physiology Institute for Zoology Chloride-mediated signal amplification in olfactory University of Heidelberg sensory neurons Im Neuenheimer Feld 230 69120 Heidelberg Most of the odor-induced receptor current in olfactory Germany sensory neurons is carried by chloride ions. We characterize A +49-6221-545661 the properties of calcium-activated chloride channels E +49-06221-545627 using a combination of electrophysiology and single-cell K [email protected] fluorescence recording (Fig. 1). We examine the role of o http://www.molecular-physiology.de these channels in stimulus-induced excitation and the molecular mechanisms of chloride accumulation which form the ionic basis of the excitatory chloride currents.

70 Stephan Frings

Chloride-dependent sensitization of nociceptors by inflammatory mediators Just like olfactory neurons, nociceptive neurons of the pain system can use chloride-dependent signal amplification (Fig. 2). But in contrast to olfactory neurons, nociceptors seem to adjust the efficiency of amplification through inflammatory mediators. We investigate this dynamic regulation of sensitivity as a novel concept for the generation of inflammatory hyperalgesia. We use an in vitro inflammation model to study the effects of inflammatory mediators on chloride homeostasis in dorsal root ganglion neurons. Applying the inflammatory agents increases net chloride uptake, raises the intracellular chloride concentration, and increases the driving force for depolarizing chloride currents through calcium-activated chloride channels. Our goal is to quantify the contribution of these chloride currents to the generation of the peripheral nociceptive signal.

Fig. 2: Chloride imaging in somatosensory neurons Using the chloride-sensitive fluorescent dye MQAE, the intracellular chloride concentration was monitored by two-photon fluorescence lifetime imaging microsopy (2P-FLIM) in an intact dorsal root ganglion. The chloride concentration within the cytosol (corresponding to the area between the two circles) differs between individual neurons. Some neurons have low (~20 mM, blue), some medium (~50 mM, green), some high (~75 mM, red) chloride levels. 2P-FLIM measurements in the presence of inflammatory mediators (PGE2, ATP, NGF, bradykinin) reveal a widespread increase of chloride levels among somatosensory neurons.

Our work, so far, shows that the two types of primary afferent neurons - olfactory sensory neurons and nociceptors - share an unusual mechanism for signal amplification. These Fig. 1: Recording calcium-dependent chloride currents neurons are able to actively accumulate chloride, and to To activate calcium-activated chloride channels, a cell is filled with a compound that releases calcium upon illumination (caged calcium) and employ an outward-directed chloride gradient to generate a fluorescent calcium indicator (fluo-5F). A series of UV flashes causes a a depolarizing chloride efflux. This chloride current boosts step-wise increase of the intracellular calcium concentration which, in the receptor potential and promotes electrical excitation. turn, increases the fluorescence signal (F). Triggered by the increasing We study proteins which mediate chloride homeostasis calcium level, chloride channels open and conduct a chloride current (I) out of the cell. In olfactory sensory neurons, this depolarizing chloride in the sensory neurons. In particular, we examine the current leads to electrical excitation. regulation of electroneutral chloride transporters

71 Stephan Frings

(NKCC1 and KCC2) which provide the driving force for Top publications the depolarizing chloride currents. Our data indicate • Reisert, J., et al. (2003). The Ca-activated Cl channel and its that the sensory cilia of olfactory receptor neurons control in rat olfactory receptor neurons. J. Gen. Physiol. 122, accumulate chloride through the activity of NKCC1 as 349-363. well as through chloride-bicarbonate exchange. This • Kaneko, H., et al. (2004). Chloride accumulation in mammalian electroneutral chloride uptake loads the cilia with chloride, olfactory sensory neurons. J. Neurosci. 24, 7931-7938. and the electrogenic chloride efflux amplifies the receptor • Bradley, J., et al. (2004). Calmodulin permanently associates potential 10-fold during odor detection. A central problem with rat olfactory CNG channels under native conditions. Nat. for research of anion-based signal amplification has yet to Neurosci. 7, 705-710. be solved: The gene that encodes the key protein - the • Kaneko, H, et al. (2006). Calmodulin contributes to gating control in olfactory calcium-activated chloride channels. J. Gen. calcium-activated chloride channel - is not identified. In Physiol. 127, 737-748. the absence of any structural infomation it is difficult • Gilbert, D., et al. (2007). Differential maturation of chloride to explore the amplification mechanism in detail. In homeostasis in primary afferent neurons of the somatosenso- collaboration with the DKFZ, we work on the molecular ry system. Int. J. Dev. Neurosci. 25,479-482. identification of this channel in olfactory receptor neurons. • Gilbert, D., et al. (2007). Caged capsaicins: new tools for the Using a proteomic approach, we have isolated the sensory examination of TRPV1 channels in somatosensory neurons. cilia from olfactory receptor neurons and characterized Chem. Bio. Chem. 8, 89-97. the membrane proteins in the ciliary membrane by mass • Funk K, et al. (2008). Modulation of chloride homeostasis by spectrometry. We are currently searching for the calcium- inflammatory mediators in dorsal root ganglion neurons. Mol. Pain 4, 32. activated chloride channel in this ciliary proteome. • Klimmeck D, et al. (2008). Calcium signaling networks in olfac- Once the chloride channel is identified, it will be possible tory receptor neurons. Neurosci. 151, 901-912. to explore anion-based signal amplification on the level of • Mayer, U., et al. (2008). Proteomic analysis of a membrane pre- channel regulation and protein-protein interaction. This paration from rat olfactory sensory cilia. Chem. Sens. 33, 145- will open new possibilities to study sensory transduction 162. in the olfactory and nociceptive systems. • Mayer U, et al. (2009). The proteome of rat olfactory sensory Thus, our group works on sensory transduction on the cilia. Proteomics 9, 322-334. levels of protein structure, ion transport, and cellular physiology. Methods range from protein biochemistry to electrophysiology and fluorescence microanalysis.

Structure of the Group Group Leader: Stephan Frings Postdoctoral fellows: Frank Möhrlen PhD-Students: Philipp Daiber, Katharina Funk, Thomas Hengl, Semir Jeridi, Nicole Ungerer, Kerstin Vocke, Clemens Waldeck Technicians : Gabriele Günther

72 Current Research Peter Gass Animal models of psychiatric disorders The role of glucocorticoid receptors in specific neu- Research Summary ronal systems for emotional behavior: a combined Using a translational approach we have established transgenic and environ- mouse models for affective disorders by two strategies: mental approach (within i) behavioral studies of transgenic mouse strains with SFB 636) mutations of genes that have been implicated in the pathogenesis of depression, e.g. glucocorticoid Glucocorticoid receptors receptors; (GRs) have been postulated ii) acute or chronic stress procedures. Our goal is to induce to play an important role in affective disorders such as de- defined alterations of the animals’ emotional behaviors pression. However, it is not known where in the brain incre- in a panel of specific behavioral tests that could serve to ased corticosteroid levels or altered GR functioning may model essential features of complex psychiatric diseases, have deleterious effects. These questions can be studied such as major depression. using conditional transgenic techniques in combination with specific mouse models for affective disorders. During Curriculum Vitae the first grant period we have characterized behaviorally Degrees: 1988 MD (Neuropathology) and neuroendocrinologically two strains of mice underex- 1996 Habilitation in Neuropathology, pressing (GR+/-) and overexpressing (YGR) glucocorticoid University of Heidelberg receptors throughout the nervous system. We could show 1983-1990: Medical School, University of Heidelberg, that GR+/- mice have a predisposition to develop typical University of Illinois and Cornell University (Ithaca, stress-induced depression-like changes (Fig. 1), while YGR N.Y., USA) mice turned out to be stress-resistant. Searching for po- 1990-1995: Residency in Neuropathology, tential molecular correlates (apart from GR) for this altered University of Heidelberg behavior we found a downregulation of BDNF in “depressi- 1996-1998: Research fellow, Dept. of Molecular Biology of the Cell (Prof. G. Schütz), DKFZ Heidelberg ve” GR+/- mice (Fig. 1) and an upregulation in YGR animals, 1999-2005: Residency in Psychiatry, CIMH which is in agreement with the so-called neurotrophin hy- 2004: Extraordinary Professor of Psychiatry, Clinical pothesis of depression. Faculty Mannheim, University of Heidelberg Since the GR gene is dysregulated during the whole since 2004: Oberarzt (Consultant) at the the Central ontogenesis in these strains, it is not clear, when and Institute of Mental Health Mannheim where (in the brain) a dysbalance of GR is important for since 2004: Head of RG Behavioral Biology of Affective affective disorders. Furthermore it is not known which Disorders at the CIMH pathophysiological and pathobiochemical alterations occur in these brain regions. These questions should Contact be investigated during the next grant period using Department of Psychiatry and Psychotherapy conditional mouse strains with specific alterations of GR Central Institute of Mental Health Mannheim expression, i.e. in the forebrain, the serotonergic system J 5, 68159 Mannheim and the noradrenergic system. To identify/rule out a A +49-621-1703-2931 E developmental role of GR expression, we will study strains +49-621-1703-2005 K [email protected] with a constitutive knockout in these brain regions as well o http://www.zi-mannheim.de as an inducible knockout. Furthermore we will investigate

73 Peter Gass

The Role of Glutamate und Glutamate Receptors in Mouse Models for Emotional Behaviors and Mood Disorders (DFG GA427/8-1, in cooperation with R. Sprengel, MPI Heidelberg) Recent studies have indicated that the neurotransmitter glutamate is involved in the pathophysiology and treatment of mood disorders (Fig. 2). The goal of this proposal is to better understand the role of glutamate in mouse models for affective disorders. We would like to find out to which extent just levels of glutamate (impaired reuptake) are important and how specific alterations of glutamate induced signalling underlie stress-related “disease states” or stress-resistance, respectively. We subject different genetically modified mice with impaired glutamate homeostasis and glutamate signalling to behavioral, neuroendocrinological, molecular Fig. 1: Glucocorticoid receptors play a causal role in the stress response and for the development of depressive behaviors. Mice with a 50% and pharmacological studies. A variety of mutant mice with reduction of GR protein levels - here always shown in red - demonstrate specific deficits in glutamate uptake (EAAT-1 and EAAT-2 a disinhibition of the HPA-system with increased stress-induced transporters) or glutamatergic neurotransmission, e.g. of corticosterone plasma levels, a non-suppression to dexamethason and a ionotropic (AMPA- and NMDA-type) glutamate receptors pathological outcome in the combined DEX/CRH test. Behaviorally these animals show increased learned helplessness in an established depression (GluR-A, GluR-C, NR-1, NR-2A) will be analysed. All mice will model, as well as a downregulation of hippocampal BDNF, a protein that be subjected to stress-induced depression models (learned has been related to pathogenesis and therapy of depression. helplessness, chronic stress) and a testing battery for emotional behaviors. We will use a step by step approach to test the “glutamate hypothesis” of mood disorders, starting with the effect of early and late environmental manipulations conventional mutant strains demonstrating the maximum (i.e. maternal care and enriched environment) in effect of gene deletions (glutamate receptors/transporters GR compromised mice to identify potential gene- genes) on emotional behavior. This will identify those ge- environment-interactions inducing alterations of nes that contribute to a behavioral phenotype. We will then emotional behaviors. As a first line of evidence we will identify the brain regions (e.g. hippocampus, forebrain) study the behavior of these mice in specific depression where the genes of interest are relevant by restricting the models (e.g. learned helplessness, chronic stress) and tests gene deletions to specific brain areas. As a proof of concept for emotional behaviors. In a second step we will analyze we will try to rescue specific behavioral phenotypes by a changes in neural plasticity that could correlate with the transgenic or alternatively a virus-mediated gene transfer. behavioral alterations by biochemical (monoaminergic We will investigate whether glutamatergic drugs that sha- systems), pathophysiological (electrophysiology) and pe emotional behavior in murine depression models have neuroanatomical (spine morphology) methods. In an altered effects in mice with genetically depleted glutamate alternative approach we plan to study in which brain receptors. We will try to identify alterations in molecular/ areas the GR is activated following environmental and biochemical/cellular signalling pathways that have been pharmacological manipulations. For this purpose we postulated for the pathogenesis or pathophysiology of de- will generate a “GR-reporter mouse”, using a construct in pression. Altogether, these experiments will improve our which a GR response element (GRE) drives the expression knowledge of the neurobiologal processes underlying the of a reporter enzyme whose activity can be measured in pathophysiology of mood disorders, and may indicate no- vitro and in situ. vel targets for antidepressant therapy.

74 Peter Gass

Top publications • Balschun, D., et al. (2003). Does cAMP response element- binding protein have a pivotal role in hippocampal synaptic plasticity and hippocampus-dependent memory? J. Neurosci. 23, 6304-6314. • Fleischmann, A., et al. (2003). Impaired long-term memory and NR2A-type NMDA receptor-dependent synaptic plasticity in mice lacking c-Fos in the CNS. J. Neurosci. 23, 9116-9122. • Zorner, B., et al. (2003). Forebrain-specific trkB-receptor knockout mice: behaviorally more hyperactive than “depressive”. Biol. Psychiat. 54, 972-982. • Strekalova, T., et al. (2004). Stress-induced anhedonia in mice is associated with deficits in forced swimming and exploration. Neuropsychopharmacol. 29, 2007-2017. • Ridder, S., et al. (2005). Mice with genetically altered glucocorticoid receptor expression show altered sensitivity for stress- induced depressive reactions. J. Neurosci. 25, 6243-6250. • Chourbaji, S., et al. (2006). IL-6 knockout mice exhibit resistance to stress-induced development of depression-like behaviors. Neurobiol. Dis. 23, 587-594. • Hellweg, R., et al. (2007). Olfactory bulbectomy in mice leads to increased BDNF levels and decreased serotonin turnover in depression-related brain areas. Neurobiol. Dis. 25, 1-7. • Chourbaji, S., et al. (2008). AMPA receptor subunit 1 (GluR-A) knockout mice model the glutamate hypothesis of depression. Faseb J. 22, 3129-3134. • Chourbaji, S., et al. (2008). Mice that under- or overexpress glucocorticoid receptors as models for depression or posttraumatic stress disorder. Prog. Brain. Res. 167, 65-77. • Vogt, M.A., et al. (2008). Suitability of tamoxifen-induced mutagenesis for behavioral phenotyping. Exp. Neurol. 211, 25-33.

Structure of the Group Fig. 2: Hypothetical scheme of abnormal glutamatergic neurotransmissi- Group Leader: Peter Gass on in mood disorders Glu=glutamate; Gln=glutamine; mGluR=metabotropic glu-tamate Postdoctoral fellows: Sabine Chourbaji , Dragos Inta, receptor; EAAT=excitatory amino acid transporter; CREB=cyclic adeno- Nada Ben Abdallah sine monophosphate response binding element protein; BDNF=brain PhD-Students: Miriam Vogt derived neurotrophic factor. Technicians: Christiane Brandwein, Christof Dormann

75 Darren Gilmour Current Research The role of chemokine-mediated tissue Mechanical regulation of migration in generating mechanosensory sensory organ morpho- organs in the zebrafish lateral line genesis in the zebrafish lateral line Research Summary (Virginie Lecaudey) The coordinated migration of groups of cells is a hallmark We have addressed the of morphogenesis. We have developed the zebrafish relationship between the lateral line primordium as a model system for the study formation of rosette-like of this poorly understood process. By combining in vivo neuromast organs and imaging with functional approaches, such as genetic the formation of sensory hair cells at their centre. Both mosaics, laser nanosurgery and small molecule inhibitors, of the processes are initiated while the primordium we have begun to address the chemical and mechanical is still migrating. Surprisingly, we have found that the cues that coordinate cell movement in the PNS. assembly of cells into organ precursors precedes the focussing of proneural genes to their centre. Furthermore, Curriculum Vitae direct mechanical perturbation of tissue migration leads Degrees: 1991 B.Sc. to predictable changes in the size and pattern of the 1996 Ph.D. neuromast organs deposited. 1991-1996: Thesis work, Gurdon Institute, Cambridge University, UK Internal FGF-signaling coordinates cell movements 1996-2004: Postdoctoral fellow, within moving tissues (Gulcin Cakan) MPI for Developmental Biology, Tübingen since 2004: Group Leader at EMBL Heidelberg Our previous work has shown that a small number of Cxcr4- expressing “leader” cells are able to guide the migration Contact of Cxcr4-deficient “followers”. In a search for potential “internal” guidance molecules, we have identified a pair of Department of Cell Biology and Biophysics EMBL FGF-family ligands (FGF3 and FGF10) that are essential for Meyerhofstr. 1 the coordinated movement of cells within this tissue. 69117 Heidelberg Deconstructing a migrating tissue by laser nanosurgery Germany (Petra Haas) A +49-6221-3878294 E +49-6221-3878306 The vast majority of migrating cell move in cohesive groups K [email protected] and the lateral line primordium is no exception. It moves as o http://www.embl-heidelberg.de/ a migrating epithelium, with cells connected via adherens and tight junctions. In order to address the role of cell-cell contacts in allowing coordinated cell movement within tissues, we have performed a comprehensive series of laser dissection experiments using a computer controlled, pulsed “laser-knife” developed by the Stelzer group at EMBL. This has revealed a suprising role for dynamic cell-cell interactions in determining directionality in this context.

76 Darren Gilmour

SDF1a coordinates tissue migration through the spatially restricted activation of chemokine receptors Cxcr7 and Cxcr4b (Guillaume Valentin, Ana Fernandez-Minan) We have previously shown that the lateral line primordium requires spatially-restricted receptors to mediate the response to an external path of the chemokine SDF-1. Cxcr4 is required in leading regions to stretch and guide the tissue along the SDF-1 source whereas Cxcr7 is expressed at the trailing edge from where neuromasts are deposited.

Current work is focused on how the combined expression of these SDF1 receptors allows these different behaviours. One approach we are currently taking is to generate transgenic reporters that allow Cxcr4 and Cxcr7 signaling to be observed in vivo.

Fig 2: Triple-staining showing the lateral line primoridum (green), Cxcr4b mRNA (blue) and Cxcr7 (red). These chemokine receptors are expressed overlapping spatial domains that correlate with different cell behaviours within the migrating tissue.

Top publications • Concha ML, et al. (2003). Local tissue interactions across the dorsal midline of the forebrain establish CNS laterality. Neu- ron 39, 423-438. • Gilmour, D., et al. (2004). Towing of sensory axons by their migrating target cells in vivo. Nat. Neurosci. 7, 491-2. • Haas, P. et al. (2006). Chemokine signaling mediates self-organizing tissue migration in the zebrafish lateral line. Dev. Cell Fig 1: Time-lapse showing rescue of lateral line primordium migration by 10, 673-80. a small number of transplanted Cxcr4b expressing “leaders” cells (red). • Lecaudey, V., et al. (2006). Organizing moving groups during morphogenesis. Curr. Opin. Cell. Biol. 18, 102-107. • Valentin, G., et al. (2007). The chemokine SDF1a coordinates Structure of the Group tissue migration through the spatially restricted activation of Cxcr7 and Cxcr4b. Curr. Biol. 17, 1026-31. Group Leader: Darren Gilmour • Lecaudey, V., et al. (2008). Dynamic FGF-signaling couples Postdoctoral fellows: Ana Fernandez-Minan, Andreea morphogenesis and migration in the zebrafish lateral line pri- Gruia, Petra Haas, Virginie Lecaudey mordium. Dev. 135, 2695-2705. PhD-Students: Gulcin Cakan-Akdogan, • Pouthas, F., et al. (2008) The Golgi apparatus and centrosome Guillaume Valentin positions depend on geometrical constraints of the substra- Technician : Andreas Kunze tum during cell migration. J. Cell. Sci. 121, 2406-2414.

77 Thomas Holstein Current Research

The origin of the metazoan nervous system Cnidarians an the origine of and body plan lower Metazoams The origin of the nervous Research Summary system and the evolution Research in our group is concerned with two major areas of multi-cellular animals in biology: Developmental neurobiology and the evoluti- (Metazoa) from single cell on of body plans. Using molecular approaches we study organisms is largely unclear these questions in basal metazoans (cnidarians) on the at present. The first animals comparative and functional level. Our principal goal is to that developed a nervous understand the basic genetic mechanisms by which the system are the cnidarians nervous system evolved, and how these developmental (coralls, sea anemones and jelly fish). These ancient processes are related to the organisation of the Bilateria. animals (>500 million years ago) have a simple nerve Of further interest are nematocytes, a highly sophistica- net and they are important to understand the origin of a ted neuronal cell type found exclusively in cnidarians. central nervous system (CNS). To trace back the evolution of the nervous system, we are analysing developmental Curriculum Vitae processes and neurogenesis in the freshwater polyp Hydra and the sea anemone Nematostella Genome Degrees: 1988 Ph.D. Vienna (Zoology) projects in Hydra and Nematostella have now revealed the 1980-1984: Postdoctoral fellow at the existence of an extensive set of conserved gene families Universities of Zurich and Munich 1985-1992: Group Leader at University of Munich and developmental pathways regulating embryogenesis 1993-1994/ throughout higher metazoans like worms, flies and 1995-1996: Visiting Professor at University of Vienna vertebrates. 1993-1997: Professor of Cell Biology, Univ. of Frankfurt 1997-2004: Full Professor of Cell Biology, TU Darmstadt Wnt and BMP signalling 1998-2004: Dean of the Faculty of Biology, TU Darmstadt The molecular nature of signalling centers (organizers) plays since 2004: Professor, Chair for Molecular Evolution and Genomics, University of Heidelberg a pivotal role in the origin and evolution of metazoan body since 2007: Dean of the Faculty of Biosciences, axes. Our lab has cloned wnt/wg genes from Hydra and University of Heidelberg Nematostella (Fig.1). The families of Wnt/Wingless secreted proteins act as short-range inducers and long-range Contact organizers in axis formation, organogenesis and tumour formation in vertebrates. In cnidarians, the canonical Wnt Department of Molecular Evolution and Genomics University of Heidelberg signalling pathway is expressed in the polyp’s oral signalling Im Neuenheimer Feld 230 center (organizer), which corresponds to the blastopore of 69120 Heidelberg metazoan embryos. In Nematostella different wnt genes Germany form staggered expression domains along the oral-aboral A +49-6221-545679 axis (“wnt-code”), governing axial differentiation, and E +49-6221-545680 neuronal differentiation of early multi-cellular animals. K [email protected] Notably, an orthologue to vertebrate dickkopf-1, -2, and -4 o http://www.zoo.uni-heidelberg.de/abt1.html genes is expressed in the body column, complementary to the expression domain of the wnt/β-catenin/tcf/brachyury genes. Dickkopf proteins, which are major Wnt antagonist

78 Thomas Holstein in vertebrates, also act in cnidarians and we propose to uncover the molecular mechanism of neurogenesis that they also promote neuronal differentiation. We are in cnidarians and to understand how it is related to the currently testing these hypotheses by using transgenic origin, evolution and patterning of neurogenesis among lines and promotor constructs for wnt, dkk, and bmp genes the metazoans (Fig. 2) as well as by testing corresponding recombinant Hydra and Nematostella proteins.

Fig. 2: The cnidarian nerve net is simple and no brain-like structures have been reported. Patterning of this ancient nervous system may have been the starting point for the formation of a central nervous system in higher animals. © Kostron and Holstein

Comparisons between vertebrates and insects have Fig. 1: The starlet sea anemone (Nematostella vectensis) is a member revealed a great deal of conservation also in the genetic of the Cnidaria. The ancient patterning system may have provided the control of neurogenesis. This suggests that the common springboard for the rapid diversification of animal body plans known as the Cambrian explosion. © Nüchter and Holstein, Nature. ancestor of Bilateria used the same set of genes to regulate neurogenesis. Yet, not all genes seem to be conserved between insects and vertebrates. Dkk proteins for example, Wnt signalling and cell adhesion which are important antagonists in the Wnt pathway and To analyse the potential link between cell signalling and cell play a role in suppressing the anti-neuronal function of adhesion during organizer formation we have identified a wnt genes, were not identified in insects and nematodes. Hydra Cadherin (HyCad) by its physical interaction with Furthermore, zic genes, which have a conserved role in Hydra β-Catenin in a yeast two-hybrid screen. We propose neurogenesis in all vertebrates downstream of Chordin that a basic function of the Cadherin-Catenin complex was and upstream of bHLH transcription factors, have not to stabilize an autocatalytic feedback loop leading to the been reported to act in neurogenesis of insects. We are definition of signalling centers. We test this hypothesis by therefore interested to learn how these regulatory genes ectopic expression and promoter analysis of both genes. are activated by the cnidarian patterning system.

The evolutionary origin of the central nervous system Imaging the dynamics of cnidarian neuronal differentiation The cnidarian nerve net is diffuse and no brain-like structures have been reported. The goal of our work is We use stable transgenes of Hydra interstitial cells to analy-

79 Thomas Holstein ze neuronal differentiation in embryos, intact polyps, and Top publications aggregates. Neuronal cells arise from pluripotent intersti- • Holstein, T.W., et al. (2003). Cnidarians: an evolutionarily con- tial stem cells that differentiate into several populations served model system for regeneration? Dev. Dyn. 226, 257- of neuronal cells (ganglion cells, sensory neurons and ne- 267. matocytes). Commitment of these cells occurs in the body • Lindgens, D., et al. (2004). Hyzic, the Hydra homolog of the zic/ column and precursor cells can migrate to the sites of final odd-paired gene, is involved in the early specification of the differentiation. This stem cell system is highly reminiscent sensory nematocytes. Dev. 131, 191-201. to the neural crest system in vertebrates. We analyze the • Technau, U., et al. (2005). High genetic complexity and mainte- behaviour of proneuronal cells along the oral aboral axis nance of non-animal genes in two basal cnidarians. Trends in in vivo and under various conditions by using confocal Genet. 21, 633-639. (COVER) laser scanning microscopy (CLSM), spinning disc confocal • Kusserow, A., et al. (2005). Unexpected complexity of the Wnt microscopy, and 2-photon microscopy. We also analyze the gene family in a sea anemone. Nat. 433, 156-60. (COVER) mechanisms of recruitment of proneuronal cells in Nema- • Rentzsch, F., et al. (2005). Glycogen Synthase Kinase 3 induces apoptosis during Hydra gametogenesis. Dev. Biol. 278, 1-12. tostella. This is particularly exciting since previous work has (COVER) shown that neuronal cells arise in more basal cnidarians • Rentzsch, F., et al. (2006). Asymmetric expression of the BMP directly from precursor cells located in the ectodermal epi- antagonists chordin and gremlin in the sea anemone Nemato- thelium. Important questions answered by this set of ex- stella vectensis: implications for the evolution of axial patter- periments are, whether precursor cells arise by asymmetric ning. Dev. Biol. 296, 375-87. cell division, whether they undergo additional proliferati- • Guder, C., et al. (2006). The Wnt code: cnidarians signal the on steps, and how the pattern of neuronal and nematocyte way. Oncogene 25, 7450-7460. recruitment is related to the axial patterning of the polyp. • Nüchter, T., et al. (2006). Nanosecond-scale kinetics of nematocyst discharge. Curr. Biol. 16, R316-8. Proteom of nematocytes • Guder, C., et al. (2006). An Ancient Wnt-Dickkopf Antagonism in Hydra. Dev. 33, 901-11. (COVER) The most conspicuous character of Cnidarians is the • Rentzsch, F., et al. W (2007). An ancient chordin-like gene in organizer formation of Hydra. PNAS 104, 3249-3254. presence of nematocysts, which are complex exocytotic • Meier, S., et al. (2007). Molecular evolution of protein folds by organelles formed inside a specialized neuronal cell single amino acid changes. Curr. Biol. 16, 173-178. type, the nematocyte (stinging cell). Nematocytes are • David, C.N., et al. (2008). Evolution of complex structures: mi- used for catching prey, and their discharge is one of the nicollagens shape the cnidarian nematocyst. Trends Genet. 24, fastest events in biology (we showed that the initial phase 431-438. of discharge is 700 nanoseconds short generating an acceleration of 6,000,000 g). The forces sustained during explosive discharge require extraordinary mechanical strength and elastic modulus in the capsular wall material. Although we made substantial progress in unravelling the molecular structure of nematocysts, the molecular Structure of the Group assembly is only partially understood. Based on our cloning Group Leader: Thomas Holstein of minicollagens and NOWA we now analyze the assembly Senior scientists: Toshitaka Fujisawa, Suat Özbek, process in a proteom project (2D-gel electrophoresis, Gabriele Petersen peptide mass fingerprinting, and tandem-MS). In parallel Postdoctoral fellows: Barbara Kostron, Ulrike Engel, Anne Kuhn, we generate antibodies and GFP-fusion constructs to Yukio Nakamura, Hiroshi Watanabe analyze the assembly process. PhD-Students: Patrizia Adamcyk, Prakasch Balasubramanian, Bianca Bertulat, Tobias Lengfeld, Robert Mättner, Martin Rittaler Technicians : Aurelia Procol, Dagmar Sealey,

80 Current Research Stefan Kins Alzheimer’s disease (AD) is Alzheimer’s Disease, APP function and characterized by a gradual transport neuronal degeneration and loss of synapses. The Research Summary microtubule associated protein Tau and the Amyloid Our research focuses on the neuronal function of the Precursor Protein (APP) were Amyloid Precursor Protein (APP) gene family and the identified as key players in molecular mechanisms underlying its intracellular the pathology of AD, but transport in neurons. Thereby we focus on alterations the reasons for the loss of of APP transport and function while aging and its synaptic connections and consequences for AD. the sequential steps of neurodegeneration in the course of AD are not understood yet. Curriculum Vitae The APP gene family consists in mammals of APP, APLP1 Degrees: 1996 M.Sc. (Diploma) and APLP2, and we could show that it functions in diverse 1999 Ph.D. biological processes, such as cell adhesion, regulation of 1996-1999: Thesis work at the MPI for brain research, Frankfurt synaptic function and axonal transport. We assume that 1999-2003: Postdoctoral fellow at the Department of deregulation of the normal APP physiological function Psychiatric research at the University of Zurich causes neurodegeneration and loss of synapses that might and the ZMBH be causative for AD and possibly explain the high risk of 2003-2006: Project Leader at theZMBH aging in AD. 2007-2008: Group Leader at the ZMBH Since 2008: Professor for Human Biology and Human Genetics at the TU Kaiserslautern

Contact University of Kaiserslautern Department of Human Biology and Genetics Erwin-Schrödinger-Straße 13 67663 Kaiserslautern Germany A +49-(0)631-205-2107 K [email protected] o http://www.uni-kl.de/wcms/bio-humanbio.html

Fig.1: Modell of APP function at the synapse The members of the APP gene family, APP, APLP1 and APLP2, form homo- and heterotypic transdimers at the synapse. Further sAPP, promoting neuronal outgrowth, is secreted in the synaptic cleft. Both features argue for an importat role of APP/APLPs in synaptogenesis and/or synaptic plasticity.

81 Stefan Kins

Specifically our research is addressing the following aims: now identified components of the active zone complex as a kinesin cargo receptor of APP transport vesicles, co- 1. Determination of the molecular mechanisms under- transporting α-secretase. As α-secretase activity abolish lying APP anterograde transport generation of the neurotoxic Aβ peptide that accumulates in plaques of AD patients, we assume that changes in the APP is anterogradely transported by very high velocity velocity of APP anterograde transport, as described for (≤10 µm/sec) and is possibly cleaved while transport along aged neurons, play a pivotal role in the etiology of AD. We the axon to the presynaptic terminal. It has been reported will determine the molecular composition of the content that APP couples the motor machinery by direct binding of the APP transport vesicle type, the precise mode of to kinesin-1. However, recent work from our and several interaction of the motor machinery and the physiological other research groups suggests that the hypothesis that consequences of alterations in transport velocity on APP APP serves as a kinesin-1 receptor and that the proteolytic pathogenic and physiological function. processing machinery responsible for generating Aβ is transported in the same vesicular compartment in axons of peripheral nerves requires revision (Back et al., 2007; Lazarov et al., 2005; Rusu et al., 2007). In a proteomic approach we

Fig. 3: Neuronal sorting of APP to dendrites and axons. Fluorescence microscopy image of a primary mouse neuron (DIV8) overexpressing mutant APP co-stained with anti-myc (green) and anti- MAP2 (red) antibodies. Arrowheads mark dendrites, arrows mark axons. Rectangular fields represent areas of quantification. For details see Back et al., 2007.

2. Characterization of the cell adhesion features of APP/APLPs Fig. 2: Model for cis- and trans-interaction of APP family proteins We found that APP/APLPs can trigger cell-cell contacts The schematic model of APP/APLP domain organization and interaction is shown. The N-terminal E1 domain is linked to a highly flexible acidic re- via homo- and heterotypic transdimerization of the gion, followed by the alternatively spliced Kunitz-type protease inhibitor ectodomains (Soba et al., 2005). Interestingly, the hetero- (KPI) domain (for APP and APLP2), the E2 domain, the juxta-/transmemb- complex of APP and APLP1 can be co-immunoprecipitated rane (TM) region, and the cytosolic domain. Based on our results, we sug- from synaptic membrane fractions, suggesting a putative gest that APP family proteins are capable of forming lateral and adhesive dimers in a homo- and heterotypic fashion. The E1 domain is crucial for function of APP/APLPs transdimerization at synaptic both, cis- and trans-interaction, while the TM region could additionally sites. To extend these studies we started now detailed contribute to lateral dimerization. immunohistological analysis with newly generated anti-

82 Stefan Kins

APP/APLPs antibodies on the ultra structural level of Top publications neuronal tissues and investigate in cell culture models its • Kins, S., et al. (2003)Hyperphosphorylation and somatoden- impact on synaptogenesis and synaptic plasticity. Further dritic localization of tau in PP2A dominant negative mutant we initiated studies addressing the consequences of APP/ mice is associated with activation of ERK and JNK signaling. J. APLPs dimerization on processing and cell migration. These Am. Path. 163, 833-843. analyses will give new insights in the physiological function • Soba, P., et al. (2005). Cell interactions are promoted by trans- of APP/APLPs and its relevance for neurodegeneration in dimerization of APP family members, arranged as homo- or AD. hetero-complexes in synaptic membranes. EMBO J. 24, 3624- 3636. • von Rotz, R.C., et al. (2005). The cytosolic ring finger protein 3. Investigations of the amyloidogenic pathway of APP dactylidin is selectively upregulated in brains of patients with processing in recycling vesicles Alzheimer’s disease. Eur. J. Neurosci. 21, 1289-1298. For the understanding of intracellular transport it is of • Lazarov, O., et al. (2005). Axonal Transport, APP, Kinesin and the Processing Apparatus: Revisited. J. Neurosci. 25, 2386-95. high impact to determine crucial sorting motifs and the • Kins, S., et al. (2006). Teasing out the tangles. Nat. Med. 12, underlying molecular machinery, including scaffolding, 764-765. regulatory and motor proteins. We found that PAT1a binds • Kuan, Y.-H., et al. (2006). The processing and anterograde trans- to the basolateral sorting signal of APP/APLPs and affects port of APP, APLP1 and APLP2 is affected by the carboxyl-ter- their subcellular localization and cleavage by α- and minal binding partner PAT1a. J. Biol. Chem. 281, 40114-40123. β-secretase (Back et al., 2007; Kuan et al., 2006). We want • Back, S., et al. (2007). In neurons APP can be transported in- to determine the molecular mechanisms underlying the dependent of any sorting signal to the axonal and dendritic effect of PAT1a on APP/APLPs transport and processing. compartment. J. Neurosci. Res. 25, 1079-1086. For this purpose we perform structural analysis and • Rusu, P., et al. (2007). Axonopathy in APP transgenic Drosophi- la depends on the NPTY motif and is paralleled by defects in investigate the influence of PAT1a on the sorting of APP/ synaptic plasticity. Eur. J. Neurosci. 25, 1079-1086. APLPs in the endocytotic pathway in neuronal cells. Using • Szodorai, A., et al. (2008). APP anterograde transport requires the yeast two-hybrid system, we identified a novel Rab5GEF Rab3A GTPase activity for assembly of the transport vesicle. J. that regulates Rab5-dependent endocytosis as a novel Neurosci. In revision interaction partner of PAT1a. Currently, we determine the • Deboer, S.R., et al. (2008). Conventional Kinesin holoenzymes molecular interplay of Rab5, the novel identified Rab5GEF, are composed of heavy and light chain homodimers. Bioche- PAT1a and APP/APLPs in the endocytotic pathway of APP/ mistry 47, 4535-4543. APLPs.

4. In vivo analyses of APP /APLPs neuronal function using viral miRNA Investigations of the loss of function of APP/APLPs in adult mice were prohibited by early postnatal death of the double- and triple knock-out mice. We have generated lentiviral constructs driving miRNA and EGFP mRNA expression Structure of the Group under a Type II polymerase promoter. After unilateral Group Leader: Stefan Kins stereotactic injection of the recombinant virus in different Postdoctoral fellows: Anita Szodorai regions of the adult brain, we will analyse those neurons PhD-Students: Simone Back, Silke Hunzelmann, with reduced protein levels of APP, APLP1 or APLP2. The Nadine Lauther, Katja Wagner knock-down cells will be analyzed by immunohistological, Undergraduates: Benjamin Gimbler electronmicroscopic and electrophysiological methods. Technicians : Sylvia Kreger

83 Joachim Kirsch Current Research Molecular cell biology of synapses Ontogeny and regulation Research Summary of glycinergic synapses in spinal cord and Fast neurotransmission is mediated by highly organized pre- and hippocampus postsynaptic protein complexes. The molecular mechanisms which establish, maintain, and alter these synaptic nano- Components of the protein machines are poorly understood. Our work focuses on translational machinery are the molecular mechanisms that regulate pre- and postsynaptic associated with juvenile protein complexes. glycine receptors and Curriculum Vitae are redistributed to the Degrees: 1987 MD (Experimental Nuclear Medicine), cytoskeleton upon aging and synaptic activity. University of Bonn Whereas adult-type glycine receptors (GlyR) are hetero- 1995 Habilitation in Anatomy oligomeric ligand-gated ion channels composed of 1978-1981: Medical School, University of Bonn α1 and β subunits, juvenile/early postnatal GlyR are 1981-1983: Dept. of Biochemical Sciences, homo-oligomeric channels composed of α2 subunits. Princeton University, USA The physiological role of juvenile/early postnatal GlyR 1983-1987: Medical School, University of Bonn 1987-1990: Postdoctoral fellow, Dept. of Neurobiology, expression for the functional differentiation of glycinergic Weizmann-Institute of Science, Rehovot, Israel synapses in the developing rat brain is not well understood. 1990-1991: Postdoctoral fellow, Dept. of Cell and Tumour It is believed that the activation of the neonatal GlyR by Biology, German Cancer Research Centre, taurine is depolarizing and accompanied by an increase 1991-1993: Postdoctoral fellow, Dept. of Neurochemistry, in intracellular Ca2+ concentrations. Moreover the Max-Planck-Institute for Brain Research, Frankfurt 1993-1995: Postdoctoral fellow, Centre for Morphology, University of Frankfurt 1995-1997: Senior scientist, Heisenberg Fellowship, Dept. of Neurochemistry, Max-Planck-Institute for Brain Research, Frankfurt 1997-1998: Endowed professorship (Schilling), Dept. of Neurochemistry, Max-Planck-Institute for Brain Research, Frankfurt 1998-2002: Professor, Chair of Anatomy and Cellular Neurobiology, University of Ulm since 2002: Professor, Chair of Medical Cell Biology, Fig. 1: Localization of eEF1 A in 28 days cultured hippocampal neurons University of Heidelberg

Contact failure of neonatal GlyR activation as a consequence of Department of Medical Cell Biology taurine deprivation can severely influence the migration Institute for Anatomy and Cell Biology of cortical neurons and thus the development of the Im Neuenheimer Feld 307 cortical architecture. In the developing rodent brain 69120 Heidelberg, Germany A +49-6221-548656 glycinergic and GABAergic neurotransmission was shown E +49-6221-544952 to be depolarizing leading to Ca2+ -influx through L-type K [email protected] channels or NMDA receptors. We could show that activation o http://izn.uni-hd.de/ of juvenile/early postnatal GlyR is crucial for the formation

84 Joachim Kirsch of postsynaptic gephyrin scaffolds, which subsequently activation. Our findings suggest a functional coupling trap adult-type glycine receptors via binding of the GlyR of eEF1A with both inhibitory and excitatory receptors, β subunit cytoplasmic pole. Moreover, the formation of possibly involving the ERK-signaling pathway. As the gephyrin scaffolds in cultured embryonic rat spinal cord monomeric GTPase eEF1A is not only involved in protein neurons requires Ca2+ -influx through L-type channels. synthesis but also thought to participate in other cellular Recently, we could demonstrate, that the GlyR α2 subunit functions such as actin bundling, cell cycle regulation, and is associated with translation eukaryotic elongation apoptosis, we investigate its cellular functions in neurons factor 1α (eEF1A) in pulldown experiments with rat with respect to dendrite morpholgy and synapse formation brain extracts. Moreover, additional proteins involved in and maintainance. translation like ribosomal S6 protein and p70 ribosomal

Fig. 2: Gephyrin clusters (green) and mCherry (red) in a transfected hippo- Fig. 3: Gephyrin clusters (green) and F-Actin (red) in cultured hippocam- campal neuron at day 21 in vitro. pal neurons after 21 days in vitro.

S6 protein kinase as well as ERK1/2 and calcineurin were Identification of novel cytomatrix proteins identified in the same pulldown approaches. Moreover, GlyR activation in young spinal cord neurons resulted in In analyzing the candidates identified in the screen we an increased phosphorylation of ribosomal S6 protein. focused on a cDNA encoding as an yet uncharacterized Immunocytochemistry showed that eEF1A and ribosomal protein, which we termed mover. Analysis of the deduced S6 protein are localized in the soma, dendrites. Their primary structure revealed that mover is a vertebrate- immunoreactivities were partially overlapping with that specific non-transmembrane protein. Biochemical data of the GlyR at synapses of cultured hippocampal and suggest that it can associate with synaptic vesicles. spinal cord neurons. Surprisingly, eEF1A immunoreactivity Upon overexpression in cultured neurons it is targeted was redistributed to the cytoskeleton in about 45% of to presynaptic terminals. Confocal immunomicroscopy neurons after 5 weeks in culture. Notably, the degree of revealed a differential localization of mover at distinct redistribution could be increased at earlier stages of in vitro subsets of CNS synapses. Whereas mover immunoreactivity differentiation by inhibition of either the ERK1/2 pathway colocalizes with presynaptic markers in the calyx of Held or GlyRs and simultaneous N-methyl-D-aspartate receptor and localizes to mossy fibre terminals in the hippocampus,

85 Joachim Kirsch it is absent from inhibitory nerve terminals in hippocampus Top publications but present at inhibitory terminals throughout the • Kins, S., et al. (2000). Collybistin, a newly identified brain-spe- cerebellar cortex. Our results suggest that mover may act cific GEF, induces submembrane clustering of gephyrin. Nat. in concert with generally expressed scaffolding proteins Neurosci. 3, 22-29. in distinct sets of presynaptic terminals. Future work • Ramming, M., et al. (2000). Diversity and phylogeny of ge- aims at the identification of mover interaction partners phyrin: tissue-specific splice variants, gene structure, and and at investigating the role of mover at distict subsets sequence similarities to molybdenum cofactor-synthesizing of synapses. Moreover, we want to elucidate by RNAi and cytoskeleton-associated proteins. Proc. Natl. Acad. Sci. experiments, which step of synaptic vesicle transport, USA 97 (18), 10266-71. fusion or recycling can be influenced by mover. • Fuhrmann, J. C., et al. (2002). Gephyrin interacts with Dynein light chains 1 and 2, components of motor protein complexes. Proteomics studies on brain peroxisomes J. Neurosci. 22, 5393-5402. • Kleinhenz, B., et al. (2005). Raver2, a new member of the hn- Peroxisomes are single membrane enclosed organelles RNP family. FEBS Lett. 579 (20), 4254-58. that catalyze a broad variety of catabolic as well as • Kuan, Y.-H., et al. (2006). PAT1a modulates intracellular trans- anabolic reactions. Peroxisomal genetic diseases, like port and processing of amyloid precursor protein APP, APLP1 X-linked adrenoleukodystophy (X-ALD) or the Zellweger and APLP2. J. Biol. Chem. 281 (52), 40114-23. syndrome are characterized by severe malformations of • Xiang, S., et al. (2006). The crystal structure of Cdc42 in complex with collybistin II, a Gephyrin-interacting guanine nucle- the CNS, indicating an important role of these organelles otide exchange factor, J. Mol. Biol. 359 (1), 35-46. in brain development. Whereas hepatic peroxisomes were • Back, S., et al. (2007). The beta-Amyloid Precursor Protein can functionally characterized in detail in the past, there is still be transported independent of any sorting signal to the axo- lack of information on the protein pattern of peroxisomes nal and dendritic compartment. J. Neurosci. Res. 85 (12), 2580- in neuronal tissue. Since peroxisomes of high purity are a 90. prerequisite of accurate protein identification, protocols • Kremer, T., et al. (2007). Mover is a novel vertebrate-specific for peroxisome isolation from brain tissue are currently presynaptic protein with differential distribution at subsets of under development using a combination of centrifugation CNS synapses. FEBS Lett. 581(24), 4727-33. techniques and free flow electrophoresis. • Bluem, R., et al. (2007). Components of the translational machinery are associated with juvenile glycine receptors and are Quantitative mass spectrometry will be used to unravel redistributed to the cytoskeleton upon aging and synaptic ac- detailed changes in the protein pattern of ALDp-knock tivity. J. Biol. Chem. 282, 37783 -37793. out and wild type mice, to obtain further insights in the • Pauly, T., et al. (2008). Activity-dependent shedding of the process disturbed in the most prevalent peroxisomal NMDA receptor glycine binding site by matrix metalloprote- disorder X-ALD. inase 3: a putative mechanism of postsynaptic plasticity. PLos ONE 3; e2681. • Dresbach et al. (2008). Molecular architecture of glycinergic synapses. Histochem. Cell. Biol. 130; 617-633.

Structure of the Group Group Leader: Joachim Kirsch Senior scientist: Jochen Kuhse Postdoctoral fellows: Thomas Dresbach, Markus Islinger, Thomas Kremer, Ralph Nawrotzki PhD-Students: Raphael Blüm, Daniel Quinones, Stefanie Schumacher Fig. 4: FLASH during Telophase. Technicians: Andrea Schlicksupp, Ingeborg Vogel

86 Current Research Georg Köhr

Cross talk between excita- Cross talk between excitatory and inhibitory tory and inhibitory synap- synapses and ion channel & signaling function ses of NMDA receptors GABA release from cerebellar molecular layer interneu- Research Summary rons can be modulated by Sensory experience affects neuronal activity, which presynaptic glutamate and/ modifies the strength or efficacy of synaptic transmission or presynaptic GABAB re- at excitatory and/or inhibitory synapses. Synaptic ceptors when perfusing the transmission can be enhanced or depressed lasting from respective agonists. Howe- milliseconds to hours or days. Our group studies short- ver, it is unclear how release and potential spillover of en- term synaptic plasticity between GABAergic interneurons dogenous transmitter lead to activation of presynaptic rein the cerebellum involving spillover of transmitter, ceptors. High frequency firing of granule cells, as observed and long-term synaptic plasticity at two hippocampal in vivo upon sensory stimulation, could lead to glutamate synapses involving NMDA receptors. In addition, we and/or GABA spillover. Using paired recordings from con- are investigating ion channel function and signaling nected stellate cells and imaging techniques (Fig. 1), we pathways of distinct NMDA receptor subtypes. study how sustained glutamatergic activity in the granule cell layer modulates GABA release in the molecular layer. Curriculum Vitae Degrees: 1989 Ph.D. in Neurophysiology Hippocampal synaptic plasticity 1996 Habilitation in Pharmacology and Long-term potentiation (LTP) or long-term depression Toxicology 1985-1989: Thesis work, Department of Neurophysiology, (LTD) of synaptic transmission are widely acknowledged MPI for Psychiatry, Martinsried and Institute of as the primary candidate manifestation of the cellular Physiology, Univ. of Cologne basis of learning and memory. Most forms of LTP and LTD 1990-1991: Postdoctoral fellow, Department of Neurology, follow a Hebbian rule, requiring coincident presynaptic Stanford University, USA and postsynaptic activity during their induction. The 1992-1996: Postdoctoral fellow, Center for Molecular Biology, presynaptic signal is thought to be glutamate release, University of Heidelberg whereas the postsynaptic signal is depolarization, allowing 1996-2000: Lecturer for Pharmacology and Toxicology, influx of calcium through NMDA receptors and/or voltage- University of Heidelberg gated calcium channels, and further supported by release 1997-2006: Project and Group Leader, MPI for Medical Research of calcium from intracellular stores. For the expression since 2001: Lecturer for Physiology, Univ. of Heidelberg of synaptic plasticity, presynaptic and/or postsynaptic since 2007: Research Group Leader, MPI for Medical Research mechanisms can be involved. Within the hippocampus, mossy fiber synapses onto CA3 Contact pyramidal neurons have unique features distinct from MPI for Medical Research other excitatory synapses. Regarding synaptic plasticity, Jahnstrasse 29, 69120 Heidelberg, Germany the most striking difference is that NMDA receptor- A +49-6221-486102 independent LTP can be induced upon high frequency E +49-6221-486110 presynaptic stimulation. Given that mossy fibers activate K [email protected] substantial NMDA receptor-mediated currents in CA3 o http://www.mpimf-heidelberg.mpg.de

87 Georg Köhr

neurons, the role of the NMDA receptors at this synapse is unclear. We investigate whether patterns of activity different from high frequency stimlulation can recruit NMDA receptors to induce synaptic plasticity at mossy fiber-to-CA3 synapses in acute slices of P21 rodents. Fig. 2 illustrates one example of spike timing-dependent plasticity (STDP; +10 ms), which induces LTP. At CA3-to-CA1 synapses, the involvement of NMDA

Fig. 2: Excitatory postsynaptic potentials (EPSPs) were evoked by mossy fiber stimulation (see scheme) in CA3 neurons of P21 rats in 10 µM bicuculline and 10 µM glycine. LTP was induced by 60 EPSP-action potential (AP) pairs. The three APs were evoked at 50 Hz and delayed by 10 ms after the evoked EPSP. The APs of the voltage traces are clipped. Note that the paired-pulse facilitation did not change after spike timing.

receptors during induction of synaptic plasticity is well established. Using genetic and pharmacological tools, our current goal is to determine whether particular NMDAR Fig. 1: (A) Scheme of a cerebellar section including electrodes for paired subtypes (see below) are recruited by different plasticity recordings (ML, molecular layer; GL, granule cell layer). (B) Test and inducing protocols. conditioned IPSCs in the postsynaptic cell (-35 mV; upper row) were evoked by paired depolarization pulses elicited in the presynaptic cell (-60 NMDA receptor subtypes: functional properties and mV; lower row). Conditioned eIPSCs were preceded by train stimulation (30 stimuli at 50 Hz). (C) A stellate cell was filled with 100 µM Oregon localization Green BAPTA-1, and granule cells were electroporated (500 µM Alexa NMDA receptors are heterotetrameric complexes Fluor 594) during train stimulation. Axonal Ca2+ transients were evoked by paired-pulse spiking (20 ms) and were acquired by two-photon composed of two NR1 and two NR2 subunits (NR2A-D) microscopy. Scale bar = 20 µm. Conditioned eIPSCs were preceded by subunits. Increasing expression of NR2A in hippocampal train stimulation (30 stimuli at 50 Hz). (C) A stellate cell was filled with neurons after birth leads to the formation of diheteromeric 100 µM Oregon Green BAPTA-1, and granule cells were electroporated NR1/NR2A and NR1/NR2B as well as triheteromeric NR1/ (500 µM Alexa Fluor 594) during train stimulation. Axonal Ca2+ transients were evoked by paired-pulse spiking (20 ms) and were acquired by two- NR2A/NR2B receptors (Fig. 3A). Despite compelling photon microscopy. Scale bar = 20 µm. evidence for triheteromeric NMDA receptors, the functional

88 Georg Köhr role of these receptors in synapses is still unknown. We are apical and basal dendrites of CA1 neurons by means of currently investigating deactivation (Fig. 3B) and peak open protein overexpression or protein knockdown using short probability of synaptic NMDA receptor subtypes in acute interfering RNAs (siRNAs). hippocampal slices of wild-type and NR2A knockout mice as these functional properties are very different between recombinant NR1/NR2A and NR1/NR2B receptors. Top publications • Köhr, G., et al. (2003). Intracellular domains of NMDA receptor subtypes are determinants for LTP induction. J. Neurosci. 23(34), 10791-10799. • Kolleker, A., et al. (2003). Glutamatergic plasticity by synaptic delivery of GluR-Blong-containing AMPA receptors. Neuron 40, 1199-1212. • Schlett, K., et al. (2004). Long-term NR2B expression in the cerebellum alters granule cell development and leads top NR2A down-regulation and motor deficits. Mol. Cell. Neurosci. 27(3), 215-226. • Pawlak, V., et al. (2005). Frequency dependent impairment of hippocampal LTP from NMDA receptors with reduced calcium permeability. EJN. 22, 476-484. • Berberich, S., et al. (2005). Lack of NMDA receptor subtype selectivity for hippocampal long-term potentiation. J. Neurosci. 25(29), 6907-6910. • Pawlak, V., et al. (2005). Impaired synaptic scaling in mouse Fig. 3: (A) Expression of two different NR2 subunits within a neuron hippocampal neurons expressing NMDA receptors with redu- can lead to the formation of three distinct NMDAR subtypes. (B) NMDA receptor-mediated EPSCs were evoked in CA1 neurons of P28 wild-type ced calcium permeability. J. Physiol. 562, 771-783. (WT) and NR2A knockout (NR2Ako) mice in 10 µM bicuculline, 5 µM NBQX • Köhr, G. (2006). NMDA receptor function: subunit compositi- and 10 µM glycine at –40 mV. (C) Schematic drawing of an excitatory on versus subcellular localization. Special Issue of Cell & Tissue synapse containing a presynaptic terminal (triangle) and a postsynaptic Research: The Synapse: Structure and Function 326, 439-446. spine carrying synaptic NMDA receptors (black paired structures). Two • Berberich, S., et al. (2007). The role of NMDAR subtypes and astrocytes ensheath the synapse and express glutamate transporters (T) charge transfer during hippocampal LTP induction. Neuro- whose activity influences the activation of perisynaptic NMDARs (dark pharmacol. 52, 77-86. gray). An extrasynaptic NMDAR is present in the dendritic shaft (light • Astori, S., et al. (2008). Sustained granule cell activity disinhi- gray). bits juvenile cerebellar stellate cells through presynaptic mechanisms. J. Physiol. 586, 575-592. Besides subunit composition, distinct NMDA receptor • Bannerman, D.M., et al. (2008). NMDA receptor subunit NR2A functions likely also depend on the spatial distribution is required for rapidly acquired, spatial working memory but of NMDA receptor subtypes within a neuron. Certain not incremental spatial reference memory. J. Neurosci. 28(14), interactions of NMDA receptor subunits with other 3623-3630. transmembrane receptors (e.g., Dopamine) and/ or intracellular signaling molecules (e.g., kinases or phosphatases) may occur at synaptic but not at peri- or Structure of the Group extrasynaptic sites (Fig. 3C). Importantly, these interactions Group Leader: Georg Köhr change during postnatal development when scaffolding Postdoctoral fellows: Simone Astori, Marie Pollard, and signaling complexes reorganize in the postsynaptic Sven Berberich densities coinciding with increasing expression of a second PhD-Students: Claudia Rauner NR2 subunit. We investigate distinct signaling pathways in MD-Students: Marina Fendel

89 Rohini Kuner Current Research Molecular mechanisms underlying chronic Mechanisms of plasticity pain at synapses between nociceptors and spinal Research Summary neurons in states of peripheral inflammation We aim at understanding molecular mechanisms underlying chronic pain resulting from long-lasting Several current projects inflammation or cancer. A major focus is laid on addressing are directed at elucidating signalling mechanisms which underlie activity-dependent novel mediators of synaptic changes in primary sensory neurons transmitting pain plasticity at synapses (nociceptors) and their synapses in the spinal dorsal horn. between primary afferents Our current work spans molecular, genetic, behavioural, and spinal neurons. In addition to transgenic approaches, electrophysiological and imaging approaches in vitro as the RNAi methodology and Adeno-associated viruses (AAV) well as in vivo in rodent models of pathological pain. are our key tools for inducing molecular perturbations in the spinal cord. We use several models of inflammatory Curriculum Vitae pain, including arthritis, to study alterations in pain Degrees: 1994 Ph.D. behaviour in vivo. Furthermore, patch-clamp recordings 2005 Habilitation and calcium imaging on spinal cord slices are employed 1994: PhD in Pharmacology und Toxicology, Dept. of for addressing potentiation of synaptic transmission. Pharmacology, College of Medicine, University of Iowa, Iowa City, USA Conditional deletion of gene encoding pain-relevant 1995-1998: Postdoctoral fellow, ZMBH, University of proteins, including receptors, kinases, G-proteins and Heidelberg; Department of Molecular ion-channels, specifically in peripheral nociceptive Neurobiology, MPI for Medical Research neurons 1998-2000: Scientist, BASF-Lynx AG, Heidelberg 2000-2001: Scientist, Pharmacology Institute, Univ. Heidelberg We utilise mouse models to test the contribution of Since 2002: Group Leader of an independent, DFG-financed important neuromediators towards learning and memory Emmy Noether-Program group, Pharmacology in pain pathways and the generation of post-inflammatory Institute, Univ. Heidelberg pain. We work with transgenic mice which lack receptors 2005: Habilitation, Medical Faculty of the of important neuromediators (e.g. Cannabinoid receptors, University of Heidelberg 2006: Professor for Pharmacology, GABA-B receptors, Glutamate receptors, Endothelin University of Heidelberg receptors, TRP channels) or their key signaling effectors (e.g. G-proteins, Protein kinases) in a temporally and Contact spatially-controlled manner using the Cre-loxP approach. Department of Pharmacology We also work on the identification and development of Im Neuenheimer Feld 366, novel therapeutic approaches for chronic pain disorders, 69120 Heidelberg, Germany with the help of viral gene delivery vectors and in vivo A +49-6221-548289 RNAi in mice. E +49-6221-548549 K [email protected] o http://www.klinikum.uni-heidelberg.de/ Pharmakologisches-Institut.106595.0.html

90 Rohini Kuner

input from the cancerous bone and this reorganization generated a neurochemical signature of bone cancer pain that is both dramatic and significantly different from that observed in mouse and rat models of chronic neuropathic or inflammatory pain. Pancreatic carcinoma is also associated with excruciating pain. Cancer pain is believed to be a unique form of pain. A distinguishing feature of cancer pain involves tumor-associated mediators which are released by tumor cells and act on sensory nerves to induce changes that elicit chronic pain. A major goal of our laboratory is to identify such tumor-associated mediators, to clarify their role in cancer pain and to develop novel therapeutic approaches to interfere with their function.

Fig. 2: Transgenic mice expressing EGFP specifically in pain-sensing nerves enable in vivo imaging in the skin of mice with chronic pain.

Structural plasticity of nociceptive nerves in disease states Structural plasticity of nerves in disease states is theme Fig. 1: On the trail of tumor-associated mediators which trigger cancer central to several of our projects. For example, in states of pain – we use the skin-nerve preparation (panel a) to study effects of bone cancer, nociceptive terminals sprout in the epider- putative tumor-associated mediators on activity of sensory nerves (panel mis of the skin adjoining the tumor. Similarly, in chronic b) and then go on to study hypersensitivity evoked by tumors in vivo pancreatitis as well as pancreas carcinoma, nerves inner- (panel c). vating the pancreas demonstrate marked hypertrophy and sprouting. Because these structural changes are usu- Molecular mediators of pain caused by bone cancer and ally studied in post-mortem tissues, it is difficult to judge tumor-nerve interactions whether they are responsible for a range of sensory ab- Pain is a frequent and devastating symptom of various normalities or whether they are mere adaptive changes forms of cancer. Various types of carcinomas and sarcomas occurring in response to the pathology and pain. In order originating from the breast, lung and prostrate metastasize to study the functional aspect of such structural changes, to bones and elicit pain. These tumour cells induce bone it is necessary to use imaging methods in living tissue. To- remodelling as well as ongoing and movement-evoked wards this end, we have established a mouse line, called pain behaviours similar to those found in patients with SNS-EGFP, expressing the live fluorescent label, EGFP so- bone cancer pain. In addition, there is a significant lely in nociceptive fibers. Furthermore, in SST2-EGFP mice, reorganization of the spinal cord that received sensory an additional mouse line which we have generated, EGFP

91 Rohini Kuner is expressed selectively in non-peptidergic nociceptors. Top publications Using such tools, we would like to address molecular me- • Dreyer, J., et al. (2004). NOS-interacting protein interacts with chanisms underlying dynamic changes in nociceptive ter- neuronal NOS and regulates its distribution and activity. J. minals in disease states. Neurosci. 24(46), 10454-10465. • Hartmann, B., et al. (2004). The AMPA receptor subunits, GluR- A and GluR-B reciprocally modulate spinal synaptic plasticity and inflammatory pain. Neuron 44, 637-50. • Kuner, R., et al. (2005). Late-onset motoneuron disease caused by transgenic expression of a functionally modified AMPA receptor subunit. Proc. Natl. Acad. Sci. USA. 102, 5826-31. • Tappe, A., et al. (2006). Regulation of motor performance and striatal function by synaptic scaffolding proteins of the Ho- mer1 family. Proc. Natl. Acad. Sci. USA 103, 774-9. Fig. 3: Pain-sensing neurons in the dorsal root ganglia display unique • Tappe A, et al. (2006) Synaptic scaffolding protein Homer1a properties and form specific synaptic connections with neurons in the protects against chronic inflammatory pain. Nat. Med. 12(6), spinal cord. Activity-dependent plasticity at this first synapse in the 677-81. somatosensory pain pathway is believed to be an important component of mechanisms leading to chronic pain. • Deng, S., et al. (2007) Plexin-B2, but not Plexin-B1, critically modulates neuronal migration and patterning of the developing nervous system in vivo. J. Neurosci. 27(23), 6333-47. Plexin-semaphorin interactions in neural development • Agarwal, N., et al. (2007). Nociceptor-specific conditional gene (joint project with the group of Prof. Stefan Offermanns, deletion reveals that cannabinoids mediate analgesia large- Pharmacology Institute, Univ. Heidelberg) ly via peripheral type 1 cannabinoid receptors. Nat. Neurosci. Semaphorins and their receptors, plexins, are key regulators 10(7), 870-9. • Michalski, C.W., et al. (2007). Cannabinoids ameliorate pain of neuronal migration and axonal pathfinding. In contrast and reduce disease pathology in caerulein-induced acute to the well-characterized plexin-A family, very little is pancreatitis. Gastroenterol. 132(5), 1968-78. known about the expression and functions of plexin-B • Tappe-Theodor, A., et al. (2007). A molecular basis of analgesic family proteins in the developing nervous system. tolerance to cannabinoids. J. Neurosci. 27(15), 4165-77. In the past, we have addressed the role of the plexin-B • Korostylev, A., et al. (2008). A functional role for Semaphorin signaling complex in growth cone behaviour and nerve 4D-Plexin-B1 interactions in epithelial branching morphoge- outgrowth in collaboration with the research group of nesis during organogenesis. Dev. 135(20), 3333-43. Stefan Offermanns. Furthermore, our expression analysis revealed that plexin-B1 and plexin-B2 and their ligand semaphorin 4D (Sema4D) are expressed at key regions at critical time points during the development of the peripheral and central nervous system. We have also addressed the role of plexin B-Sema4D interactions Structure of the Group in epithelial-mesenchymal interactions during kidney Group Leader: Rohini Kuner development. Our current projects are focussed on Postdoctoral fellows: Martina Kurejova, Ceng Luo, Anke Tappe- characterizing mouse mutants lacking plexin-B1 or Theodor, Eszter Paldy, Manuela Simonetti, plexin-B2 for defects in the development of the peripheral Nitin Agarwal, Daniel Vardeh and central nervous system. PhD-Students: Vijayan Gangadharan, Alexandra Kulkova, Kiran Kumar Bali, Lucas Vicuna, Christian Njoo, Deepitha Selvaraj, Jianning Lue Technicians: Dunja Baumgartl-Ahlert, Tamara Djuric, Hans-Joseph Wrede, Eva Reiss

92 Current Research Thomas Kuner Molecular structure and Structure and function of synapses function of central nerve terminals Research Summary Our work is aimed at understanding the molecular Synaptic communication mechanisms of synaptic transmission, mainly focusing on between neurons relies on presynaptic nerve terminals. We employ a multidisciplin- the transfer of all-or-none ary approach ranging from molecules to behavior: signals: presynaptic action molecular perturbation, viral gene transfer, genetically potentials are translated encoded indicators, high-resolution fluorescence into graded synaptic microscopy, electron microscopy, 3D analyses, currents on the postsynaptic quantitative fluorescence imaging, electrophysiology side. Mainly two scaling factors define the efficacy of this and behavior. translation: the magnitude of the response generated by a single action potential and the frequency-dependent Curriculum Vitae modulation of this translation process. Most synapses Degrees: 1998 MD 1992-1997: Thesis work with Peter Seeburg at the ZMBH, University of Heidelberg 1998-2000: Postdoctoral fellow training with George Augustine at Duke University, Durham, USA 2000-2006: Group Leader at the department of Bert Sakmann, MPI for medical research, Heidellberg since 2006: Professor at the University of Heidelberg, Institute for Anatomy and Cell

Contact Department of Medical Cell Biology Institute for Anatomy and Cell Biology Im Neuenheimer Feld 307 69120 Heidelberg Germany A +49-6221-548678 E +49-6221-544952 K [email protected] o http://ana.uni-hd.de/ag-kuner

Fig. 1: Calyx of Held presynaptic nerve terminal visualized by viral expression of GFP. Width of image ~40 µm.

93 Thomas Kuner

respond to repeated action potentials by a transient and the active zones of synaptic terminals. short-lasting reduction of synaptic scaling, known as short-term depression (STD). This synaptic filtering has a We study synaptic transmission in a giant nerve fundamental impact on neuronal computation. Current terminal of the rat auditory brain stem known as the evidence suggests that the depletion of synaptic vesicles calyx of Held. The synapse formed by the calyx with its (SV) and the rate of recruitment of recycled or new SVs to postsynaptic neuron, the principal cell of the medial the active zone (AZ) determines the extent of STD at an nucleus of the trapezoid body (MNTB), is the only individual synapse. Therefore, the dynamics of membrane central synapse of vertebrates from which simultaneous trafficking events control an important parameter of pre- and postsynaptic electrophysiological recordings synaptic transmission and neuronal communication. can be routinely achieved. We combine molecular biology & genetics, electrophysiology, fluorescence imaging, immunohistochemistry, three-dimensional reconstructions and high-pressure quick-freeze electron microscopy to study these questions in an integrated multi-dimensional approach.

Neuronal chloride signaling imaged with a genetically encoded indicator The polarity of GABAergic or glycinergic synaptic transmission is controled by the transmembrane gradient of Cl- and the resting membrane potential. At a typical membrane potential of -70 mV, GABA and glycine yield hyperpolarizing responses at low intracellular concentrations of Cl-, but depolarizing responses at high concentrations of Cl-. Hence, the control of intracellular Cl- can have a fundamental influence on GABAergic and glycinergic neurotransmission by defining the polarity of the postsynaptic response. Using Clomeleon, a genetically Fig. 2: Wrapped donuts - a geometrical organizational principle encoded indicator for Cl-, we study the spatial and found in adult calyces of Held. A: P21 calyx with GFP-synaptophysin- temporal distribution of Cl- in hippocampal neurons and labeled synaptic vesicles (entire shown structure ~ 20 µm diameter). B: the impact of Cl- gradients and local accumulation on arrangement of SVs after EM 3D reconstruction (diameter of structure ~ 1 µm). GABAergic synaptic transmission. Clomeleon is expressed in selected neuronal subtypes by transgenic or viral techniques. We use quantitative ratiometric confocal and To understand the molecular basis of this process it is crucial two-photon imaging combined with electrophysiology to to know more about the reactions that translocate synaptic study intracellular Cl- dynamics in neurons. vesicles from the reserve pool to the AZ and prepare them for Ca2+-dependent release of neurotransmitter (NT) From molecules to behavior: neuronal mechanisms of quanta. In fact, very little is known about the precise spatio- odor discrimination temporal organization and the identity of protein-protein interactions underlying these processes in neurons. Our Lateral inhibition is a prominent mechanism generating goal is to systematically investigate protein function in the contrast enhancement in sensory systems. In the olfactory context of synaptic vesicle translocation to, and priming at, system, lateral inhibition is thought to be mediated by a

94 Thomas Kuner synaptic interaction between mitral cells (MC) and granule Top publications cells (GC) of the olfactory bulb. Dendrites of MCs and GCs • Wimmer, V. C., et al. (2004). Targeted in vivo expression of pro- are connected by the reciprocal synapse, a specialized teins in the calyx of Held. Pflug. Arch./Europ. J. Physiol. 449, bidirectional contact capable to act as a receiver and 319-333. sender of synaptic signals. Thus, glutamate release • Abraham, N.M., et al. (2004). Maintaining accuracy at the ex- from MC dendrites can translate into a graded response pense of speed: stimulus similarity defines odor discriminati- involving either GABA release from the same terminal on time in mice. Neuron 44, 865-876. (reciprocal inhibition), from nearby ones (local lateral • Dübel, J., et al. (2006). Two-photon imaging reveals somato- inhibition), or even from all terminals of a GC (global lateral dendritic chloride gradient in retinal ON-type bipolar cells ex- inhibition). The contribution of these mechanisms to odor pressing the biosensor Clomeleon. Neuron 49, 81-49. • Wimmer, V.C., et al. (2006). Donut-like topology of synaptic discrimination in the mouse is only poorly understood. We vesicles with a central cluster of mitochondria wrapped into study the effect of specific molecular manipulations at the membrane protrusions: a novel structure-function module in reciprocal synapse of GCs on odor discrimination in mice. the adult calyx of Held. J. Neurosci. 26, 109-116. Acute targeted genetic perturbations (ATGp) will be used • Pond, B.B., et al. (2006). The chloride transporter Na(+)-K(+)-Cl- to specifically interfere with molecular targets in granule cotransporter isoform-1 contributes to intracellular chloride cells of the olfactory bulb. The behavioral consequences of increases after in vitro ischemia. J. Neurosci. 26:1396-1406. perturbations will be assessed with an odor discrimination • Berglund, K., et al. (2006). Imaging synaptic inhibition in mice test (Abraham et al., 2004). After behavioral testing expressing the chloride indicator, Clomeleon. Brain Cell. Biol. identified relevant perturbations, the physiological 35, 207-235. • Tappe, A., et al. (2007). A molecular basis of analgesic toleran- consequences will be examined with electrophysiological ce to cannabinoids. J. Neurosci., 4165-4177. and imaging techniques on the level of individual GCs. • Krieger, P., et al. (2007). Synaptic connections between layer 5B pyramidal neurons in mouse somatosensory cortex are independent of apical dendrite bundling. J. Neurosci. 27,11473- 11482. • Kuner, T., et al. (2008). Photolysis of a caged peptide reveals rapid action of N-ethylmaleimide sensitive factor before neurotransmitter release. PNAS 108, 347-352. • Groh, A., et al. (2008). Driver or coincidence detector: modal switch of a corticothalamic giant synapse controlled by spontaneous activity and short-term depression. J. Neurosci. 28, 9652-9663.

Structure of the Group Group Leader: Thomas Kuner Postdoctoral fellows: Nixon Abraham, Robert Renden, Anna Dondzillo Fig. 3: Expression of Clomeleon in hippocampal CA1 neurons of a PhD-Students: Christian Kempf, Christoph Körber, Clomeleon indicator mouse line. Daniel Nunes, Darius Schwenger, Bhavana Shrivastava, Francisco Urra, Maryia Vasileva Technicians: Heinz Horstmann, Michaela Kaiser, Claudia Kocksch

95 Siegfried Mense Current Research Neurobiological mechanisms of chronic Sensitisation of dorsal horn muscle pain neurones by subthreshold potentials Research Summary Electrophysiological experi- Animal experiments in vivo address the role of neuroacti- ments in the rat have shown ve mediators in peripheral and spinal muscle nociception, that under normal circum- as well as the potential involvement of spinal glial cells stances many dorsal horn in the development of chronic muscle pain. Further, the neurones react to input from studies aim at comparing neurobiological mechanisms muscle nociceptors with of chronic pain in a locomotor muscle with those in low subthreshold membrane back muscles. potential changes, only. Intramuscular injections of the neurotrophin nerve growth factor, for example, activate Curriculum Vitae many nociceptive muscle afferents but at the level of Degrees: 1973 MD spinal dorsal horn neurones, this activation elicits mainly 1979 Habilitation subthreshold potentials. These potentials do not evoke 1966-1971: Thesis work at the Dept. of Nuclear Medicine, subjective pain sensations but nevertheless sensitise University of Bonn central nociceptive neurones. The intracellular recordings 1971-1978: Postdoctoral fellow in Electrophysiology at the in vivo give a more complete picture of the spinal wiring of Dept of Physiology, University of Kiel muscle nociceptors compared to the conventional extra- 1978-1979: Assistant professor at the Dept. of Physiology of cellular microelectrode techniques. These data are the the University of North Carolina at Chapel Hill 1979-1985: Group Leader at the Department of Physiology, University of Kiel 1985-2008: Professor at the University of Heidelberg, Institute for Anatomy and Cell Biology

Contact Chair of Neuroanatomy Research Division Neurobiology Center for Biomedicine and Medical Technology Mannheim (CBTM) Medical Faculty Mannheim, University of Heidelberg Theodor-Kutzer-Ufer 68167 Mannheim Fig.1: Stimulating effects of nerve growth factor (NGF) on dorsal horn Germany neurones. A +49-621-383-9713 A) Original registrations of a dorsal horn neurone responding with sub- E +49-621-383-9949 threshold excitatory postsynaptic potentials (EPSP) but no action K potentials (APs) to an intramuscular NGF injection. Likewise, noxious [email protected] pressure (Nox. p.) stimulation of the muscle evoked EPSPs only (upper o http://www.ma.uni-heidelberg.de/inst/cbtm/ panel). B) Number of neurones responding to intramuscular NGF injection. Only a few dorsal horn neurones responded with occasional APs (and EP- SPs) to the NGF injection, a greater number showed EPSPs only. The mean AP frequency after NGF was extremely low (2.6 APs per min).

96 Siegfried Mense basis of a more detailed understanding of spinal processes contribution of spinal microglial cells to the development induced by tonic, subthreshold membrane potential chan- of chronic muscle pain. Microglia is assumed to play an ges in nociceptive dorsal horn neurones. The understan- important role in chronic pain states. In a rat model of ding of central sensitisation evoked by tonic subthreshold chronic muscle inflammation, morphological alterations input provides new insight in neurobiological mechanisms of spinal microglial cells can be evaluated by quantitative that underlie chronic muscle pain syndromes that often OX-42 immunohistochemistry. OX-42 is a specific marker develop without a perceived tissue lesion. of spinal microglia. In behavioural experiments, the effects of intrathecally applied minocycline - a specific blocker of Role of spinal microglia in central sensitisation microglia activation - on pain-related behaviour caused by muscle inflammation are studied. Simultaneously, Previous studies of our group had shown that muscle processes at the neuronal level are investigated with inflammation does not only lead to marked changes intracellular recordings of nociceptive dorsal horn in the responsiveness of dorsal horn neurones but also neurones. In these experiments, the influence of induces morphological and functional alterations in neuroactive substances released by microglial cells- or of spinal astrocytes. One aim of the current research is blockers of these substances - is examined. to employ immunohistochemical, behavioural, and electrophysiological methods to investigate the

Fig. 3: In animals with a chronic muscle inflammation, the threshold of the hindlimb withdrawal response was determined with mechanical pressu- Fig. 2: Quantitative evaluation of microglial OX-42 immunoreactivity (IR) re stimuli. The inflammation led to a persistent drop in the pressure pain in intact rats and in rats with a chronic muscle inflammation. threshold. Continuous intrathecal application of 200 µg minocycline per A) As areas of interest, six fields (179 µm x 133 µm each) were defined, 3 day with an implanted osmotic pump significantly increased the lowered in the superficial dorsal horn and 3 in the deep dorsal horn. Within the pain threshold (*, P < 0.05). These data show that the myositis-induced rectangles, the OX-42 immunoreactive (ir) areas and their boundary drop in pain threshold is largely due to an activation of microglia lengths were measured. B) OX-42 IR in the deep dorsal horn visualised by the green fluorescent marker Cy2 (lamina V, GS muscle inflamed). Neurobiological mechanisms of chronic low back pain C) 8-bit grey image of OX-42 IR (same figure as shown in B). D) Quantitative evaluation of immunostaining. The threshold level for Although the prevalence of chronic pain in low back immunostaining against background was defined as the mean back- muscles is very high, there are practically no basic science ground grey value in unstained areas between the fluorescent microglia (white rectangle in C) plus three standard deviations. data on possible mechanisms of this type of pain. Most of E) In animals with an inflamed muscle (inflamed), the mean boundary the current knowledge of mechanisms of muscle pain was length of the immunoreactive (ir) areas were significantly shorter in obtained in studies on locomotor muscles of the extremities. the deep dorsal horn (Eb; ***, P < 0.001), whereas in these region the We are now performing systematic animal experiments ir area was not reduced (Ea). The decrease in boundary length is probably due to a retraction of the cell processes during inflammation- to investigate the neurobiological mechanisms of low induced activation of the microglial cells. back pain and compare the data with those obtained in

97 Siegfried Mense experiments on locomotor muscles. The central nervous Top publications connections of low back muscles are examined by injecting • Mense, S., et al. (2003). Lesions of rat skeletal muscle after local retrograde tracers into supraspinal centres and visualising block of acetylcholinesterase and neuromuscular stimulation. nociceptive dorsal horn neurones with input from the J. Appl. Physiol. 94, 2494-2501. low back using the cFos reaction. We just completed the • Hoheisel, U., et al. (2004). Acidic pH and capsaicin activate me- first systematic study of the responsiveness of dorsal horn chanosensitive group IV muscle receptors in the rat. Pain 110, neurones to noxious stimulation of low back muscles. In 149-157. these experimens, single neurones were recorded from • Hoheisel, U., et al. (2005). Excitatory and modulatory effects of with extracellular recording techniques. inflammatory cytokines and neurotrophins on mechanosen- sitive group IV muscle afferents in the rat. Pain 114, 168-176. • Hoheisel, U., et al. (2005). The possible role of the NO-cGMP pathway in nociception: Different spinal and supraspinal action of enzyme blockers on rat dorsal horn neurones. Pain 117, 358-367. • Hoheisel, U., et al. (2007). Sensitization of rat dorsal horn neurones by NGF-induced subthreshold potentials and low-frequency activation. A study employing intracellular recordings in vivo. Brain Res. 1169, 34-43. • Lambertz,D., et al. (2008). Influence of a chronic myositis on rat spinal field potentials evoked by TTX-resistent unmyelini- nated skin and muscle afferents. Eur. J. Pain 12, 686-695. • Taguchi, T., et al. (2008). Dorsal horn neurons having input from low back structures in rats. Pain 138, 119-129 Fig. 4: Lumbar dorsal horn neurone retrogadely labelled from the peri- • Chacur, M., et al. (2008). Role of spinal microglia in myosi- aquaeductal gray matter (PAG) with Fluorogold (Aa). The same neurone tis-induced central sensitisation: an immunhistochemical also exhibited cFos-immunoreactivity after noxious stimulation of low and behavioural study in rats. Eur. J. Pain doi: 10.1016/j. back muscles (Ab). B) Responses of a nociceptive dorsal horn neurone ejpain.2008.11.008. to noxious stimulation of deep tissues in the low back. The neurone also responded to noxious pressure applied to the paw. Each vertical line in the original recordings is an action potential. The data show that neurones with input from the low back project to the PAG and exhibit a marked input convergence from various tissue types.

Structure of the Group Group Leader: Siegfried Mense Postdoctoral fellows: Ulrich Hoheisel, Toru Taguchi PhD-Students: Natalia Bode, Viola John, Jonas Tesarz Scientists: Daniela Lambertz Technician: Beate Quenzer, Marion Schmitt, Marina Szymbara, Claudia Tolliver

98 Current Research Andreas Meyer-Lindenberg Neurogenetic risk mecha- Translational neurogenetics of psychiatric nisms of schizophrenia disorders We study the impact on genetic risk variants (COMT, PP- Research Summary P1R1B, DISC1, RGS4, etc.) on Many severe psychiatric disorders and complex neural function and cogni- behaviours are highly heritable. We investigate genetic tion using multimodal neu- variation associated with risk for disorders such as roimaging. A recent focus of schizophrenia, depression and autism and behavioural that work is the investigati- phenomena such as attachment or impulsive violence in on of epistatic interactions a translational approach, with an emphasis on multimodal (COMT x GRM3, COMT x RGS4), relating multiple genetic neuroimaging to identify neural systems mediating variants to neural phenotypes, and the study of neural me- genetic risk. The ultimate goal is the construction of a chanisms predictive of therapeutic response and course neural risk architecture of the studied target behaviour as in a longitudinal perspective (Meyer-Lindenberg et al., guide for the discovery and evaluation of novel treatment 2005b; Meyer-Lindenberg et al., 2005c; Meyer-Lindenberg strategies. et al., 2006b; Goldman et al., 2007; Tan et al., 2007) Curriculum Vitae Neurogenetic risk mechanisms of depression and Degrees: 1991 MD (Neurochemistry) anxiety 1999 Habilitation University of Giessen Here, we focus on mechanisms of gene x environment 1984 -1991: University of Bonn (Medical School) 1989: Research fellow, Cornell University, New York interactions and the neural circuits mediating association 1997-2001: Visiting Associate Research fellow, National of genetic variation with trait anxiety. Genetic variants Institute of Mental Health, Clinical Brain Disorders studied here include 5-HTTLPR, MAO vntr, and BDNF. Branch, Bethesda, USA (Pezawas et al., 2005). 2001-2005: Staff scientist, NIMH, Clinical Brain Disorders Branch, Co-director of the Imaging Core Facility Neural mechanisms of complex social behaviours under since 2007: Professor and Chairman of Psychiatry and genetic control Psychotherapy, University of Heidelberg, Clinical Director of the Department of Psychiatry and Many aspects of social behaviour are highly heritable, are Psychotherapy and Director of the Central key components of severe psychiatric disorders that are Institute of Mental Health themselves heritable, and are essential for reproductive fitness. Consequently, we have launched a program to Contact investigate genetic contributions to the human social brain. Department of Psychiatry and Psychotherapy We have identified neurogenetic mechanisms contributing Central Institute of Mental Health to violence, impulsivity, attachment and personality traits. J 5, 68159 Mannheim In parallel, we are interested in studying social phenomena Germany that show a high degree of conservation across species, A +49-621-17032001 such as social hierarchies.(Meyer-Lindenberg et al., 2005a) E +49-621-17032005 K [email protected] o http://www.zi-mannheim.de

99 Andreas Meyer-Lindenberg

plasticity in the prefrontal cortex (PFC) in vivo is quantified using a combination of repetitive transcranial magnetic stimulation (rTMS) and functional neuroimaging to derive detailed measurements of cortical physiology at an intermediate phenotype level. We expect that systems- level correlates of plasticity will be impaired in carriers of a functional val66met variant in the BNDF gene and modulated by variants in the genes COMT, NRG1, and GRM3. (Pezawas et al., 2004).

Top publications • Meyer-Lindenberg, A., et al. (2004). Neural basis of genetically Fig. 1: (a) Neural mechanisms of hypersociability in WS (Meyer-Lindenberg determined visuospatial construction deficit in Williams syn- et al., 2005a). Amygdala activation for socially relevant face stimuli (top drome. Neuron 43, 623-31. row) and less relevant scene stimuli (bottom row) stimuli, rendered on • Meyer-Lindenberg, A., et al. (2005). Midbrain dopamine and normal coronal MRI. First column: normal controls (NC), Second column: prefrontal function in humans: interaction and modulation by participants with WS (WS). Subjects with WS show decreased activation to COMT genotype. Nat. Neurosci. 8, 594-596. social fear stimuli, but increased signalling to non-social stimuli, mirroring • Pezawas, L*, et al. (2005). 5-HTTLPR polymorphism impacts their anxiety profile. (b) Neural circuits for amygdala regulation and social human cingulate-amygdala interactions: a genetic suscep- function under genetic control impacted in WS (Meyer-Lindenberg et tibility mechanism for depression. Nat. Neurosci. 8, 828-34 al., 2006a): both orbitofrontal cortex (OFC) and cingulate interact with amygdala in healthy controls, but OFC is disconnected in WS. (c) structure (*shared first authorship). (left) and functional connectivity (right) affected by genetic variation in • Meyer-Lindenberg, A., et al. (2005). Neural correlates of gene- 5-HTTLPR (Callicott et al., 2005). Carriers of the short allele show relative tically abnormal social cognition in Williams syndrome. Nat. volume reductions in subgenual cingulate and amygdala, and reduced Neurosci. 8, 991-993. connectivity of amygdala to subgenual cingulate, delineating a feedback • Kirsch, P., et al. (2005). Oxytocin modulates neural circuitry for circuit for fear extinction and social cognition. (d) structural (left, using social cognition and fear in humans. J. Neurosci. 25, 11489- voxel-based morphometry) and functional results (right, during an 11493. emotional faces matching task) show an impact of genetic variation in • Meyer-Lindenberg, A., et al. (2006). Neural mechanisms for ge- MAO-A on amygdala, cingulate and orbitofrontal volume and function (Meyer-Lindenberg et al., 2006c). Volume is relatively reduced in carriers netic risk for violence and impulsivity in humans. Proceedings of the low expression allele implicated in risk for impulsive violence. of the Nat. Acad. Sci. USA 103, 6269-6274. Amygdala activation is increased, while activation of regulatory cingulate • Meyer-Lindenberg, A., et al. (2007). Genetic evidence impli- and orbitofrontal regions is decreased. (e) the prosocial hormone cating DARPP-32 in human fronto-striatal structure, function oxytocin modulates amygdala activation in healthy humans (Kirsch et al., and cognition. J Clin Invest. 1, 117(3), 672-682. 2005). Stimuli and display as in (a). Top: placebo, Bottom: oxytocin.. • Zink, C.F., et al. (2008). Know Your Place: Neural Processing of Stable and Unstable Social Hierarchy in Humans. Neuron Apr Genetic modulation of neural plasticity 24;58(2), 273-83. • Meyer-Lindenberg, A., et al. (2008): Genetic variation in AVP- In this new project, the goal is to identify the functional R1A linked to autism predicts amygdala activation and perso- correlates, neurochemical mechanisms and the impact of nality traits in healthy humans. Mol Psychiat. May 20. genomic variation on experimentally induced prefrontal • Esslinger, C., et al. (2009) Neural mechanisms of a genome- cortical plasticity in the human. Short-term cortical wide significant psychosis variant. Science. 324, 605

100 Current Research Hannah Monyer

Molecular mechanisms Neuronal synchrony and plasticity underlying synchronous activity in the central Research Summary nervous system The studies of this lab are directed at the identification of genes critically involved in the generation of synchronous oscillation A number of compelling in neuronal networks, at the functional analysis of different studies and computational interneuronal populations and at the characterization of the role simulations data provide of AMPA and NMDA glutamate receptor subtypes in different evidence that networks of forms of plasticity. inhibitory neurones play a pivotal role in the generation Curriculum Vitae of certain forms of oscillations that arise as a network Degrees: 1983 MD property. Oscillatory activity in different frequency ranges 1994 Habilitation has been proposed to be critical for a number of cognitive 1976-1982: Medical School, Thesis work, University of tasks, including object representation, learning and Heidelberg 1983-1984: Resident at the University Hospital for Psychiatry, memory. Department of Child Psychiatry, Mannheim The goal of our studies is to identify ‘key’ molecules in 1984-1986: Resident at the University Hospital for Pediatrics, GABAergic interneurones that underlie oscillatory activity Department of Pediatric Neurology, Lübeck and that are involved in controlling synchronous firing 1986-1987: Postdoctoral research fellow, Stanford University of ensembles. Our studies entail analysis from the single Medical Center, Department of Neurology, EEG cell level to the network activity in vitro (acute slice Laboratory, Stanford, USA preparation) and in vivo. Genetically modified mice with 1987-1989: Postdoctoral research fellow, Stanford University altered expression of critical genes (e.g. AMPA receptors, Medical Center, Department of Neurology, NMDA receptors, connexins) in GABAergic interneurones Neurology Research Laboratory, Stanford, USA 1989-1994: Postdoctoral research fellow, Center for Molecular are a critical tool for subsequent electrophysiological Biology, University of Heidelberg studies to investigate synchronous network activity in 1994-1999: Hermann-and-Lilly-Schilling-Foundation cortex and hippocampus in the freely moving mice. These Professor at the Center for Molecular Biology, investigations are further complemented by behavioural University of Heidelberg studies. since 1999: Head of the Department of Clinical Neurobiology, IZN and Neurological University, University Hospital Characterization of different interneuronal populations Heidelberg since 2002: Speaker of the Graduate College Programme Given the large diversity of GABAergic interneurones GK 791 (based on the presence of certain parameters, for instance chemical markers, morphological criteria, connectivity), Contact present projects aim at the identification of GABAergic Department of Clinical Neurobiology subpopulations of neurones (e.g. parvalbumin-, University Hospital of Neurology Im Neuenheimer Feld 364 somatostatin-, calretinin-positive cells). To this end 69120 Heidelberg, Germany transgenic mice are produced in which these neuronal A +49-6221-562401 subpopulations are labelled using the in vivo marker green E +49-6221-561397 fluorescent protein. The subsequent electrophysiological K [email protected], [email protected] studies on fluorescent neurones in these mice should aid o http://www.KlinNbio.uni-hd.de

101 Hannah Monyer in identifying the GABAergic cell types involved in different AMPA receptor interacting proteins and synaptic forms of network oscillations. plasticity In transgenic mice that express EGFP in a particular subset AMPA receptors, another subtype of the glutamate recep- of GABAergic interneurones, we have found that unlike the tor family, mediate most of the fast excitatory transmission vast majority of GABAergic interneurones that are gene- in the vertebrate central nervous system. They are critical in rated embryonically, certain GABAergic interneurones determining the strength of transmission at glutamatergic continue to be generated after birth. Postnatal neurogenesis synapses, and tight regulation of their function allows for of GABAergic interneurones that migrate into the cortex use-dependent and input-specific adaptations of synaptic and hippocampus is a novel form of plasticity. The analysis is strength. Their function is influenced by composition, based on anatomical and electrophysiological techniques posttranslational modifications and by protein-protein as well as imaging studies of migrating neurones. interactions. We have identified a novel brain specific Furthermore, these mice are used for gene expression AMPA receptor interacting protein (AIP47) by mass analysis in EGFP-labelled neuronal populations by means spectrometry of AMPA receptor complexes and we are of laser dissection microscopy. currently studying the role of this protein in regulating AMPA-receptor mediated synaptic transmission and plasticity of glutamatergic synapses.

Fig. 1: A:Example of an in vivo electrophysiological hippocampal recording in transgenic mice. B:Example of behavioral analysis of transgenic mice.

Fig. 2: A: Schematic drawing of neuroblasts migrating from different migratory stream to the cortex and the olfactory bulb. NMDA receptors neuronal plasticity and vulnerability B: Picture of fluorescent neuroblasts in the dorsal migratory stream of the The NMDA receptor, a subtype of the glutamate receptor 5HT3-EGFP mouse. family, is critical for the induction of different forms of plastic changes in the brain. Anatomical and functional characterization of NMDA receptors subtypes has revealed that pyramidal neurones co-express the two NMDA receptor subtypes NR2A and NR2B. The differential developmental regulation of these two NMDA receptor subtype expression with respect to brain areas and cell types exerts an important function in the change of neuronal plasticity during brain maturation. Projects pertaining to this research programme aim at cell type-specific ablation of the NR2B subunit to study the function of the ‘young’ receptor form in the adult brain. The analysis of the genetically modified mice is performed using electrophysiological and behavioural studies.

102 Hannah Monyer

Top publications Structure of the Group • Blatow, M., et al. (2003). A novel network of multipolar burs- Group Leader: Hannah Monyer ting interneurons generates theta frequency oscillations in Senior Scientist: Andrei Rozov neocortex. Neuron 38, 805-817. Postdoctoral fellows: Julieta Alfonso, Rosa Arribas Prat, • Bruzzone, R., et al. (2003). Pannexins, a family of gap junction Antonio Caputi, Elke Fuchs, proteins expressed in brain. Proc. Natl. Acad. Sci. USA 100, Kaneko Hiroshi, Anne Herb, 13644-13649. Alexey Ponomarenko, Corentin Le • Monyer, H., et al. (2004). Interneuron Diversity series: Molecu- Magueresse, Jakob von Engelhardt lar and genetic tools to study GABAergic interneuron diversity PhD-Students: Christina Göngrich, Vogt Angelika, and function. Review. Trends Neurosci. 27, 90-97. Aleksandar Zivkovic • Christie, J. M., et al. (2005). Connexin36 mediates spike syn- Scientists: Maria Blatow chrony in olfactory bulb glomeruli. Neuron 46, 761-772. Undergraduates: Seda Ballikaya, Michael Barbe, • Traub, R.D., et al. (2005). Transient depression of excitatory Panagiotis Bargiotas, Christine Bark, synapses on interneurons contributes to epileptiform bursts Christina Bocklisch, Irena Brinkmann, during gamma oscillations in the mouse hippocampal slice. J. Golovko Tatiana, Khrulev Sergey, Neurophysiol. 94, 1225-1235. Jennifer Lee, Attila Racz, Olga Voinova • Fuchs, E. C., et al. (2007). Recruitment pattern of parvalbumin- Technicians: Ulrike Amtmann, positive interneurons determines hippocampal function and Regina Hinz-Herkommer, Marlies Kaiser, associated behavior. Neuron 53, 591-604. Konstantin Khodosevich, • von Engelhardt, J., et al. (2007). Functional characterization of Irmgard Preugschat-Gumprecht, intrinsic cholinergic interneurons in the cortex. J. Neurosci. 27, Andrij Sergyienko, 5633-5624. Heike Xoumpholphakdy • Khodosevich, K., et al. (2007). Gene expression analysis of in vivo fluorescent cells. PloS ONE (2)11, e1151. • Caputi, A., et al. (2008). Two Calretinin-positive GABAergic cell types in layer 2/3 of the mouse neocortex provide different forms of inhibition. Cerebral Cortex, doi: 10.1093/cercor/ bhn175 • von Engelhardt, J., et al. (2008) Contribution of hippocampal and extra-hippocampal NR2B-containing NMDA receptors to performance on spatial learning tasks. Neuron. 60, 846-860

103 Ulrike Müller Current Research

Functional genomics of neurodegenerative Alzheimer´s disease (AD) diseases; Alzheimer´s disease, physiological ist the most common and pathological function of APP family neurodegenerative disorder and is characterized by proteins synaptic disfunction, neuronal loss and cognitive Research Summary decline. The major lesions Our research focuses on the molecular mechanisms of found in the brains of AD synaptic transmission disorders and the pathogenesis patients are neurofibrillary of neurodegenerative diseases, in particular Alzheimer´s tangles and neuritic plaques disease. Our aim is to unravel the physiological and that are mainly composed pathogenic function of key genes and mechanisms that of the β-amyloid peptide (Aβ) derived via proteolysis trigger disease. from the amyloid precursor protein APP. APP is a single pass transmembrane protein that is processed in two Curriculum Vitae different ways: α-secretase cleaves APP within the Aβ Degrees: 1985 Diploma, University of Munich region, thereby precluding Aβ formation and releasing 1989 Dr. rer. nat., University of Munich the APPsα ectodomain; in the amyloidogenic pathway 1999 Habilitation in Molecular Biology APP is sequentially cleaved by β- and γ-secretase, leading University of Zurich to Aβ formation. Whereas the mechanisms governing Aβ 1989-1991: Postdoc, Medical School, generation have been intensely studied, the physiological University of Manchester, UK role of APP and of its numerous proteolytic fragments 1991-1997: EMBO long term fellow and Group Leader, and the question of whether a loss of these functions Institute for Molecular Biology, University of Zürich contributes to AD are still unknown. 1997-2004: Independent Research Group Leader, (C3), Head of the Neurogenetics group, MPI for Brain Research, Neurochemistry, Frankfurt Knockout mice with individual or combined gene Since 2005: Professor for Functional Genomics, Department deficiencies of APP-family proteins of Bioinformatics/ Functional Genomics, IPMB, Determining the in vivo functions of APP in mammals is University of Heidelberg complicated by the presence of two APP-related genes, APLP1 and APLP2. APP and APLPs share two conserved Contact domains in the extracellular region (E1 and E2) and one in Institute for Pharmacy and Molecular Biotechnology - IPMB the cytoplasmic domain, whereas the β-amyloid peptide University of Heidelberg is lacking in APLPs. Thus, functional redundancy may Im Neuenheimer Feld 364 compensate for the loss of essential gene functions, e.g. 69120 Heidelberg Germany in knockout (KO) models. Indeed, by generating various A +49-6221-546717 KO mutants we could demonstrate that the extensive E +49-6221-545830 structural similarities between APP and APLPs are also K [email protected] reflected at the functional level. Mice in which APP, APLP1, o http://www.uni-heidelberg.de/institute/fak14/ipmb/ or APLP2 is inactivated are viable and APP-KO mice revelaed reduced brain and body weight, reduced grip strength, altered locomotor activity, increased susceptibility to seizures and a defect in spatial learning and LTP. In contrast

104 Ulrike Müller to the viable single mutants, combined APLP2‑/‑APP‑/‑ and have become major targets of therapeutic intervention APLP2‑/‑APLP1‑/‑ double mutants die shortly after birth it is of primary importance to elucidate the physiological indicating that APP family proteins serve redundant relevance of APP processing and to understand the functions that are essential for viability (Heber et al., 2000). specific functions of the respective cleavage product Whereas the brains of double knockout animals exhibit for both physiology and pathophysiology. To this end no obvious morphological defects, triple mutants lacking we recently started a reverse genetic analysis of APP the entire APP gene family showed cranial dysplasias functional domains. We replaced the endogenous APP resembling human type II lissencephaly (Herms et al., 2004). locus by gene targeted alleles and generated two lines of Within affected areas neuronal cells from the cortical plate knockin mice that exclusively express APP deletion variants migrated beyond their normal positions and protruded corresponding either to the secreted APP ectodomain into the marginal zone and the subarachnoid space. Thus, (APPsα) or to a C-terminal truncation lacking the YENPTY APP/APLPs play a critical role in neuronal adhesion and interaction motif (APPΔCT15) (Ring et al., 2007). Most positioning. This role in cell adhesion is also supported by importantly, we demonstrated that in both APP knockin data from a recent collaborative study demonstrating that lines the expression of APP N-terminal domains either APP family proteins form cis- and trans-dimers involved largely attenuated or completely rescued the prominent in cellular adhesion and may thus play a role in synaptic deficits of APP knockout mice, such as reductions in brain differentiation/function (Soba et al. 2005). Collectively, our and body weight, grip strength deficits, alterations in data revealed an essential role for APP-family members in circadian locomotor activity, exploratory activity, and the normal brain development and early postnatal survival. impairment in spatial learning and LTP. Taken together Work is in progress to circumvent early lethality and our data suggest that APPsα is sufficient to mediate the assess functions postnatally by generating tissue specific physiological functions of APP assessed by these tests knockouts. (Ring et al., 2007). Ongoing experiments will show whether APPsα might also be sufficient to rescue defects underlying the lethal phenotype of APP‑/‑APLP2‑/‑ mutants.

The role of APP and its fragments for neuronal morphology and synaptic function Recently, a role of APP and APLP2 at the neuromuscular synapse has been described. Thus, ongoing work in the lab is directed to assess whether APP/APLP deficiency is also associated with related defects of neuronal morphology and/or synaptic function within the CNS. To this end we are analyzing organotypic hippocampal cultures of knockout and knockin mice with regard to morphology and (in Fig. 1: Frontal section of a triple KO mouse at E 17.5 exhibiting a prominent collaboration) for their electrophysiological characteristics protrusion (P) of the cortical plate. Ectopic neurons completely disrupt the cortical plate (CP); and neuroblasts are shifted into the marginal zone (basal synaptic transmission, synaptic plasticity). (MZ). (adapted from Herms et al., 2004) Role of APP-dependent gene expression for Alzheimer In vivo analysis of APP functional domains disease Another level of functional diversity may result from The proteolytical processing of APP is very similar to the complex proteolytic processing of APP and its APLP that of Notch and the APP intracellular domain AICD has homologues by several secretases leading to diverse extra- been suggested to function as a transcriptional regulator. and intracellular APP/APLP fragments. As these secretases Nevertheless, the nature of the relevant target genes is

105 Ulrike Müller still under debate (see e.g. Pardossi-Picard et al., 2005 Top publications and Hebert et al., 2006). Using a microarray/qPCR based • Heber, S., et al. (2000). Mice with combined gene knockouts approach we identified novel differentially expressed genes reveal essential and partially redundant functions of Amyloid (e.g. involved in cytoskeletal remodeling, endocytosis, cell precursor protein family members. J. Neurosci. 20, 7951-7963. adhesion and neurotransmitter systems). In this context • Harvey, R., et al. (2004). GlyRα3: an essential target for spinal we are particularly interested to clarify whether these PGE2-mediated inflammatory pain sensitization. Science 304, target genes are directly regulated at the mRNA level via 884. AICD acting in a Notch-like manner, or are more indirectly • Herms, J., et al. (2004). Cortical Dysplasia Resembling Human affected by the absence of APP-family proteins. Type 2 Lissencephaly in Mice Lacking all Three APP-Family Members. The EMBO J. 23, 4106 – 4115. Regulation and molecular pathology of the inhibitory • Grimm, M. O. W., et al. (2005). Regulation of cholesterol and glycine receptor sphingomyelin metabolism by amyloid-β and presenilin. Nat. Cell. Biol. 7, 1118-1123. Glycine receptors (GlyRs) are ligand activated chloride • Soba, P., et al. (2005). Homo- and hetero-dimerization of APP channels composed of α- and β subunits forming a family members promotes intercellular adhesion. EMBO J. 24, pentamer. GlyRs are involved in the control of spinal 3624-3634. motor and sensory pathways, but little was known about • Hebert, S.S., et al. (2006). Regulated intramembrane proteoly- the biological roles of different GlyR subtypes. Recently sis of amyloid precursor protein and regulation of expression of putative target genes. EMBO reports 7, 739-45. we showed that GlyRα3 subunits are distinctly expressed • Hirzel, K., et al. (2006). Hyperekplexia phenotype of glycine in superficial laminae of the dorsal spinal cord, the first receptor α1 subunit mutant mice identifies Zn2+ as an essen- site of synaptic integration in the pain pathway (Harvey tial endogenous modulator of glycinergic neurotransmission. et al., 2004). At this site, glycinergic neurotransmission is Neuron 52, 679. inhibited by prostaglandin E2 (PGE2), a pivotal mediator of • Ring, S, et al. (2007).The secreted APPsα domain is sufficient inflammatory pain sensitization. By generation of GlyRα3 to rescue the anatomical, behavioral, and electrophysiologi- knockout mice we could show that this receptor subtype cal abnormalities of APP deficient mice. J. Neurosci. 27, 7817- plays a crucial role in spinal nociceptive processing. Mice 7826 deficient in GlyRα3 not only lacked the inhibition of • Tamboli, I., et al. (2008). Loss of γ-secretase function impairs glycinergic neurotransmission by PGE seen in wild-type endocytosis of lipoprotein particles and membrane choleste- 2 rol homeostasis. J. Neurosci. 28, 1207-12106 mice, but also showed a reduction in pain sensitization induced by spinal PGE2 injection or peripheral inflammation (Harvey et al., 2004). Thus, GlyRα3 may provide a novel molecular target in pain therapy.

Structure of the Group Group Leader: Ulrike Müller Postdoctoral fellows: Viola v. Bohlen, Mikhail Filippov, Jakob Tschäpe Fig. 2: Transverse section through wild type spinal cord. Double labeling PhD-Students: Dorothee Aydin, Sabine Ring, shows that GlyRα3 (green) is restricted to the dorsal horn, and gephyrin Sascha Weyer (red) is expressed throughout the gray matter. Technicians: Diana Bundschuh, Julia Gobbert, (adapted from Harvey et al., 2004) Michael Neumann, Frankziska Stöckling

106 Current Research Christof Niehrs It is now commonplace that Molecular Embryology principles guiding embryonic development in humans Research Summary and in animals are very simi- The Division of Molecular Embryology is studying lar on a molecular level and mechanisms regulating cell differentiation. The aim is that genes involved in deve- to characterize molecular mechanisms relevant for the lopment also play a role in formation of the body axis in frogs and mice. To this human disease. The Division end, we are identifying developmental control genes of Molecular Embryology is and investigating how these are regulated and what studying mechanisms regu- their functions are. Our special interest is devoted to lating embryonic cell diffe- investigating the molecular characteristics of a region rentiation in Xenopus and mouse. Our aim is to characte- of the embryo called Spemann organizer. Another focus rize molecular mechanisms relevant for the formation of is the systematic analysis of gene activity during early the embryonic body axis. To this end, we identify deve- embryonic development lopmental control genes, investigate what their biological role and biochemical mode of action is and how they are Curriculum Vitae regulated. We specifically address the following two main topics: I. Molecular mechanisms of Wnt/β-catenin signal- Degrees: 1985 Diploma (M.Sc.) 1990 Ph.D. ling during early vertebrate development. Elucidation of 1997 Habilitation the functions of of Dickkopf (Dkk) proteins, a class of Wnt 1986-1990: Thesis work at EMBL, Heidelberg inhibitors, is a major focus. II) Molecular mechanis of em- 1990-1993: Postdoctoral fellow, University of California, L.A., bryonic reprogramming, with the aim to elucidate mole- USA cular mechanisms regulating embryonic pluripotency. since 1994: Group Leader of the division: “Molecular embryology” at the German Cancer Center (DKFZ), Heidelberg

Contact Department of Molecular Embryology German Cancer Research Center (DKFZ) Im Neuenheimer Feld 581 69120 Heidelberg Germany A +49-6221-424690 E +49-6221-424692 K [email protected] o http://www.dkfz.de/en/mol_embryology

Fig. 1: Xenopus embryos at 4-cell stage.

107 Christof Niehrs

Mechanisms of Wnt signaling in early vertebrate gastrula. Together with other data this led to the proposal development of perpendicular activity gradients of Wnt and BMPs, which

regulate early CNS patterning (Fig. 2). The Wnt/β-catenin pathway plays an eminent role in We previously identified an important regulator of development and disease. We showed that in Xenopus Spemann’s organizer, dickkopf1(dkk1), member of a new embryogenesis, Wnt/β-catenin signaling plays an family of secreted proteins. Dkk1 encodes a Wnt inhibitor important role in antero-posterior patterning of the and its functions in embryonic head development early central nervous system. Our results demonstrated a (Fig. 3). While the gene was initially characterized as a posteriorizing gradient of Wnt signalling, which regulates dose-dependently positional information from the forebrain to spinal cord. Importantly, we revealed an endogenous antero-posterior gradient of Wnt/β-catenin signalling in the presumptive neural plate of the Xenopus

Fig. 3: Day 17 mouse embryos wild type (left) and dkk1 null mutant.

developmental regulator, dkk1 is gaining increasingly medical attention, since it has been implicated in bone physiology and myeloma by others. We currently study the role of Dickkopf genes and their receptors Kremen and LRP6 during Xenopus and mouse development, including CNS. Towards this end we identify interacting proteins of these proteins and elucidate their role in Wnt signalling and early development. This led to the recent discovery of Rspondins and Casein kinase 1 gamma as novel Wnt regulators, which are also involved in CNS patterning. Fig. 2: Double-gradient model for A-P and D-V axial patterning The model shows how perpendicular activity gradients of Wnts and bone Reprogramming morphogenetic proteins (BMPs) regulate head-to-tail (A-P) and dorsal– ventral (D-V) CNS patterning. The colour scales of the arrows indicate Another line of research in our laboratory is devoted to the signalling gradients; arrows indicate the spreading of the signals. the question of reprogramming i.e. the artificial process Patterning begins at gastrula stages, but for clarity, it is depicted in an of transformation of a somatic into a pluripotent cell. The early amphibian neurula. The formation of head, trunk and tail requires increasing Wnt activity. Anterior and posterior are to the left and right, interest is two-fold. First, by identifying regulatory genes respectively. that control the state of pluripotency we hope to learn

108 Christof Niehrs how the differentiated vs. pluripotent state is regulated. Top publications Second, this knowledge may be used to generate • del Barco Barrantes, I., et al. (2003). Dkk1 and noggin coopera- human isogenic pluripotent stem cells for regenerative te in mammalian head induction. Genes Dev. 15, 2239-2244. medicine. Towards this end we have developed partial • Böttcher, R., et al. (2004). The transmembrane protein XFLRT3 embryonic reprogramming of entire somatic cells by forms a complex with FGF receptors and promotes FGF signal- using Xenopus egg extracts. Screening for factors required ling. Nat. Cell Biol. 6(1), 38-44. for reprogramming identified the chromatin remodelling • Kazanskaya, O., et al. (2004). R-Spondin2 Is a Secreted Activa- ATPase BRG1. tor of Wnt/beta-Catenin Signaling and Is Required for Xeno- Furthermore, we have identified a molecular mechanism pus Myogenesis. Dev Cell. 7(4), 525-34. that is involved in demethylation and re-activation of • Davidson, G., et al. (2005). Casein kinase 1 couples Wnt receptor activation to the cytoplasmic signal transduction. Nature pluripotency genes, such as oct4. This mechanism involves 438(7069), 867-72. the stress response gene Gadd45, which acts by recruiting • Barreto, G., et al. (2007). Gadd45a promotes epigenetic gene DNA repair to sites of demethylation. Methylated cytosines activation by repair-mediated DNA demethylation. Nature are excised and replaced by unmethylated nucleotide, 445(7128), 671-5 thereby leading to demethylation. This is a novel epigenetic • Bilic, J., et al. (2007). Wnt induces LRP6 signalosomes and pro- mechanism of gene activation. We currently study the role motes dishevelled-dependent LRP6 phosphorylation. Science of this process during early Xenopus development and we 316(5831), 1619-22. screen for factors that mediate the site specific targeting of Gadd45 to sites of demethylation in the genome.

Structure of the Group Group Leader: Christoph Niehrs Scientists: Gary Davidson, Mathias Gierl, Andrei Glinka, Emil Karaulanov , Olga Kazanskaya, Bisei Ohkawara, Cristina Sirrenberg-Cruciat PhD-Students: Kristina Ellwanger, Wolfram Gruhn, Ya-Lin Huang, Malte Paulsen, Andrea Schäfer, Jinlong Shen, Lilian Sitter, Yuliya Sytnikova Technicians: Gabriele Döderlein, Ursula Fenger,

109 Sabina Pauen Current Research Early childhood brain and cognitive Categorization in infancy development Categorization in infancy is based on bottom-up Research Summary processes (i.e. perceptual Using behavioural as well as neurophysiological abstraction) as well as on data (heart rate, ERPs), we explore lower and higher top-down processes (i.e. order cognitive functions (e.g. intermodal perception, memory activation). As work categorization, memory formation, causal and functional conducted in our lab has understanding, social cognition) in early childhood shown, top-down processes development. Furthermore, we are interested in the play an important role even relation between emotional and cognitive self-regulation at a preverbal age. Infants perform a global-to-basic- and how it is influenced by early childhood experience. level shift, with more abstract categories (i.e. animates vs. inanimates) developing earlier than basic-level categories Curriculum Vitae like dogs, cats, chairs or tables (Pauen, 2002a). As indicated Degrees: 1988 Diploma (M.Sc) by follow-up studies, these categorical distinctions reflect 1992 Ph.D. conceptual knowledge (Pauen, 2002b). So far, only 1999 Habilitation behavioural measures have been used to study infant 1985-1988: University of Marburg (Psychology Diploma) categorization. In our lab, we also use heart rate measure 1990-1991: University of Giessen - DFG Research assistant (Elsner, Pauen & Jeschonek, 2006) and ERP-measures (see 1991-1993: University of Tübingen - Research assistant Figure 1) to validate behavioural data. Corresponding 1993-1994: Cornell University - DFG Postdoctoral fellow studies demonstrate that even the brains of 7-month- 1994-1999: University of Tübingen - Research assistant 2000-2001: University of Magdeburg Leader DFG-Junior Research Group Since 2002: University of Heidelberg Full professor (C4)

Contact Department of Psychology University of Heidelberg Hauptstrasse 47-51 69117 Heidelberg Germany A +49-6221-547269 E +49-6221-547326 K [email protected] o http://www.psychologie.uni-heidelberg.de/ae/entw/index. html

Fig. 1: Infant prepared to conduct an EPR-Study on categorization.

110 Sabina Pauen old infants respond differently to animal and vehicle Yet another line of research focuses on how infants use pictures. Furthermore, we found that 12-month-olds functional knowledge to guide their categorization show different brain responses when looking at pictures of artefacts. Based on the finding that 12-month-olds of artefacts or animals accompanied by matching vs. non- categorize objects differently after their functional use matching sounds. Taken together, our studies provide first has been demonstrated than without a corresponding neuropsychological evidence for the early emergence of a demonstration (Träuble & Pauen, 2007) we now explore knowledge-based animate-inanimate distinction. in more detail the conditions that support this kind of functional learning (social cues, objects cues; e.g. Elsner & Causal and functional reasoning in infancy Pauen, 2007). This line of research focuses on the question what kind of knowledge about objects infants acquire during the first Relation between early cognitive development and year of life. One of the key attributes of animates is self- language acquisition initiated movement. Using newly developed experimental In early lexical acquisition, noun understanding emerges paradigms, we have shown that even 7-month-olds are quite early. To better understand how preverbal able to attribute the cause of a given motion to animates categorization and noun comprehension interact, we rather than inanimates. When watching an unfamiliar investigate the impact of labelling on categorization animal moving around together with a ball, they attribute performance in infants 7 to 12 months of age. Results reveal the cause of this motion to the animal rather than the ball that in very young infants, nouns increase general attention (see Figure 2). for objects, but do not change the pattern of categorization, whereas for older infants who are about to learn basic- level categorical distinctions, nouns can indeed help to make these distinctions within a given global domain. Furthermore, we are interested in studying whether early preverbal categorization skills have prognostic value for later intelligence. Investigating the relation between verbal and non-verbal measures of intelligence for preschoolers, we found that non-verbal categorization skills in a basic- level task at 11 months of age and noun comprehension at 12 months of age (assessed via parental questionnaires) covariate. Furthermore, recent data from our German Language Development Study reveals that verbal and non-verbal measures of intelligence in 4- to 5-year olds are more closely related that originally thought.

Development of analogical reasoning This line of studies focuses on the beginnings of analogical reasoning in human infants. We approach this issue by conducting comparative studies with infants, monkeys and chimpanzees in collaboration with primate labs at the Fig. 2: Infant participating in a study on early causal reasoning: It watches MPI in Leipzig as well as the monkey-lab at the University an animal and a ball rolling together. Later, looking time measures will of Rome. Our studies and collaborations focus on three reveal whether the motion has been attributed to the animal or the ball. related topics: (1) intermodal integration of dynamic

111 Sabina Pauen displays, (2) perceptual understanding of the sameness Top publications and differentness relation, and (3) tool-use as well and • Goswami, U., et al. (2005). The Effects of a ‘Family’ Analogy on relational mapping. Class Inclusion Reasoning by Young Children. Swiss J. Psychol. Infants are shown dynamic displays presenting one 64, 115-124. upward and one downward moving circle, accompanied • Schöppner, B., et al. (2006). Encoding action roles in meaning- either by a tone of rising or descending pitch. Using a ful social interaction in the first year of life. Infancy 9, 389-411. preferential-looking paradigm, we will find out whether • Pauen, S. (2006). Infant cognitive psychology and the under- intermodal relational mappings typically found in adults standing of learning processes. Europ. Psychol. 11, 263-265. (i.e. an upward movement is associated with a rising pitch) • Pauen, S. (2006). Was Babys denken. München: Beck-Verlag, are rooted in basic intermodal perceptual integration 2nd edition. • Elsner, B., et al. (2006). Physiological and behavioral parame- of the brain from birth on, or can better be explained by ters of infants’ categorization: Heart rate and duration of exa- learning processes occurring later in life. In similar ways, mining across trials. Dev. Sci. 9, 551-556. we plan to explore other intermodal relations, such as the • Elsner, B. et al. (2007). Social learning of artefact function in 12- relation between luminance and pitch. and 15-month-olds. Europ. J. Dev. Psychol. 4, 80-99. One very basic perceptual relation that plays a crucial role • Träuble, B. et al. (2007). The role of functional information for in categorization is the relation of sameness / differentness. infant categorization. Cognition 105, 362-379. Existing animal studies suggest that monkeys are capable • Pauen, S. et al. (2008). Psychologie des Säuglings- und Klein- of judging sameness and differentness, when provided kindalters. In Psychologie des Säuglings- und Kleinkindalters. with at least 8 to 16 pictures that either look identical or Enzyklopädie der Psychologie, Vol 4, M. Hasselhorn, et al., eds. (Göttingen, Germany: Hogrefe Verlag),pp. 67-126. different from each other. This leaves open the question • Pauen, S. et al. (2008). Neurobiologische Grundlagen der Ent- whether they really understood the underlying concept or wicklung. In Entwicklungspsychologie, R. Oerter, et al., eds. solved the task by simply recognizing that the elements of (Weinheim, Germany: Beltz PVU), 6. Ed., chapt. 3, pp. 67-84. some complex display either showed a graphical pattern • Pauen, S. et al. (2008). How to investigate the concept of con- or did not. Ruling out this methodological problem, cepts. J. Anthrol. Psychol. 19, 32-35. the present study will compare monkey and infant performance in a response-match-to sample task, using only two simple graphic figures per display that are either the same or different from each other. Analogical reasoning plays a crucial role in problem solving. Together with Rome we designed a tool-use study in which monkeys and infants learn to use one of three sticks, differing in the appearance of their handles as well as in the length of the sticks, to push some reward out of a tube. Only the longest of the three sticks reaches the reward in the tube – thus providing an effective tool. Following a training phase, the infants and animals will get a new set Structure of the Group of tools. All three sticks have the same length as before, but the handles are exchanged. It will be tested whether Group Leader: Sabina Pauen the infants / monkeys focus on the functionally relevant Postdoctoral fellows: Birgit Elsner, Gudrun Kane, Janna Pahnke, feature (i.e. stick length) or the perceptually salient cue (i.e. Birgit Träuble, Eva Vonderlin the handle) when choosing their tool. Scientists: Lysett Babocsai, Deise Desch, Alenka Hribar, Susanna Jeschonek , Anna Ropeter, Cornelia Schrauf, Isa Valentina, Andrea Wittke

112 Current Research Gabriele Elisabeth Pollerberg

Our research is focussed Growth and orientation of axons in the on the question how axons developing nervous system (the long extensions sent out by developing and Research Summary regenerating nerve cells) We are interested in the cellular and molecular processes are able to protrude, extend, underlying the formation of axonal connections during and navigate towards their embryonic development of the higher vertebrate central targets where they will nervous system. We study interactions of elongating axons ultimately form synapses. and the growth cones at their tips with the environment At the tip of each axon is a and the transformation into directed growth, in particular motile, sensory structure, the growth cone, which is able the roles of cell adhesion molecules (CAM), microtubule to sense growth permissive and non-permissive cues binding proteins (MBP) & intracellular signalling. in its environment and to react accordingly. Important molecules in the environment as well as in the growth Curriculum Vitae cone membrane are cell adhesion molecules (CAMs) which hence act both as external guidance cues and Degrees: 1979 Diploma (M.Sc.) University of Konstanz 1986 Ph.D. (Dr.rer.nat. University of Heidelberg) as axonal receptors by interacting with each other. The 1995 Habilitation (University of Tübingen) external signals are transformed into an adequate growth 1982-1986: Thesis work, University of Heidelberg cone reaction (turn towards or turn away; stop, retraction 1986-1987: Postdoctoral fellow; EMBL Heidelberg or advance) by intracellular signalling cascades which 1988-1996: Group Leader (MPI for Developmental Biology, ultimately act on the local stability of the cytoskeleton. Tübingen) The growth cone cytoskeleton thus is the structure 1996-1998: Professor (Head of Dev. Biology Dept., which defines the future growth direction of the axon: University of Giessen) The cytoskeleton is stabilized on one side of the growth 1997: Visiting Professor, National Institute for cone and destabilized on the other which leads to a local Basic Biology, Okazaki, Japan 2002-2003: Visiting Professor, Institute of Zoology and School collapse and protrusion formation, respectively, so that the of Medicine, Beckman Center axon in the following turns to the side where the growth Stanford University, USA cone is net stabilized. This differential stabilisation is since 1999: Professor and Head of Developmental Neurobiolo- mainly modulated by microtubule binding proteins (MBPs) gy Dept., University of Heidelberg which are capable of enhancing microtubule growth and stability. Contact In particular, our research concentrates on the roles of two Department of Developmental Neurobiology CAMs of the immunoglobulin superfamily, DM-GRASP Institute of Zoology and NrCAM, for extending axons since they are both axon- University of Heidelberg specific and expressed during early axon growth. CAMs Im Neuenheimer Feld 232 not only mediate adhesion but also elicit intracellular 69120 Heidelberg signalling pathways; this way, CAMs have an downstream Germany impact on cytoskeletal dynamics. We thus also analyse the A +49-6221-546370 E functional impact of MBPs, in particular MAP1B and APC, on +49-6221-546375 K [email protected] the stability of the microtubule system in the growth cone o http:// www.zoo.uni-heidelberg.de/gep/ and the consequences for axonal navigation. Moreover,

113 Gabriele Elisabeth Pollerberg we study the modulators of the MBPs which activate or laser scanning microscopy 3D reconstructions (Fig. 1c). inactivate their microtubule stabilizing properties and Time lapse analysis in retina flat-mount cultures moreover thereby mediate between plasma membrane receptors revealed that the RGC axons grow slower and meander and cytoskeleton. more on their way to the optic nerve head if DM-GRASP Our model system is the developing visual system of the or NrCAM is blocked. Together these findings show for the avian embryo. The retina is the best accessible part of first time that these two CAMs have an important impact the central nervous system since it is basically the only on axonal growth and navigation. structure not covered by bone. Due to its extra-uterine development, the avian embryo is a higher vertebrate embryo which can be readily manipulated and observed in the intact egg over long time periods. In addition, the avian embryo model system provides a series of culture techniques which allow for the study of navigating axons in their natural histotypic environment, e. g. in eye organ culture or in retina flat-mount culture. We concentrate on extension and navigation of retinal ganglion cell (RGC) axons which are the only axons to leave the retina; they dive into the optic nerve head and project to the optic tectum of the midbrain.

Roles of the CAMs DM-GRASP and NrCAM for axonal growth and pathfinding The spatio-temporal expression analyses which we performed for DM-GRASP and NrCAM in the embryonic Fig. 1: Impact of CAMs on axons. (a) NrCAM offered as substrate lanes visual system revealed that both CAMs are exclusively (N) is preferred over laminin (L) by RGC axons. (b) Blocking of DM-GRASP present on extending, fasciculating RGC axons which form results in RGC axons crossing the optic fissure (OF) and growing into the the innermost optic fibre layer in the retina and regional opposite retina side. (c) In contrast to the slim growth cones focussed towards the OF (upper panel), blocking of NrCAM (lower panel) leads to subsets in the optic nerve, chiasm, optic tract, and optic perpendicularly probing complex growth cones. tectum, indicating their function for early axon growth and navigation. Using an in vitro assay system mimicking Roles of microtubule binding proteins MAP1B and APC this restricted presence of DM-GRASP and NrCAM in the growth cone environment by narrow substrate lanes Growth cone behaviour such as advance, pause, turn, and coated on glass cover slips, we could show that both retraction and thereby the growth direction of the axon CAMs are able to guide RGC axons (Fig. 1a). Moreover, is largely determined by the microtubule system which both CAMs enhance axon growth on laminin substrate, is modulated in its dynamics by MBPs. In growth cones indicating that the presence of these CAMs could speed induced to turn at a substrate border, we could show that up RGC axons in vivo. Both CAMs could be demonstrated a special phosphorylated form of the axon-specific MBP to contribute crucial to the correct routing of RGC axons microtubule-associated protein 1B (MAP1B) is only found into the optic nerve head by inhibition experiments in the in the stable region of the growth cone, i. e. in the side of intact retina (Fig. 1b). Also the focussing of the RGC growth the future growth direction. Kinase Cdk5, which performs cones towards the optic fissure and the limitation of their this type of MAP1B phosphorylation, is present in the probing activities away from the correct growth direction entire growth cone. The activator of Cdk5, P35, however, is depends on the presence of the CAMs as visualized by only found in the stable part of the growth cone; thereby

114 Gabriele Elisabeth Pollerberg a local activation of MAP1B phosphorylation can lead to a Top publications differential stabilisation/collapse of growth cone regions. • Hernandez-Sanchez, C., et al. (2003). Upstream AUGs in emb- We could thus demonstrate a crucial role of MAP1B and its ryonic proinsulin mRNA control its low translation level. EMBO modulators Cdk5/P35 for growth cone turning responses J. 22, 5582-5592 and thereby axonal navigation. We also investigated • Avci, H.X., et al. (2004). Role of cell adhesion molecule DM- how APC, another MBP present in the growth cone (Fig. GRASP in growth and orientation of retinal ganglion cell axons. 2a), affects the dynamics of microtubules and thus the Dev. Biol. 271, 291-305. behaviour of the growth cone. We neutralized domains of • Hahn, C.M., et al. (2005). Role of cyclin-dependent kinase 5 the large, multifunctional protein APC by chromophore- and its activator P35 in local axon and growth cone stabilizati- assisted laser inactivation (micro-CALI) in one half of the on. Neurosci. 134, 449-465. growth cone. Inactivation of the N-terminal domain (which • Zelina, P., et al. (2005). The cell adhesion molecule NrCAM is crucial for growth cone behaviour and pathfinding of retinal is crucial for APC’s dimerisation and hence conceivably ganglion cell axons. Dev. 132, 3609-3618. for microtubule bundling/stabilisation) results in growth • Koester, M.P., et al. (2007). Adenomatous polyposis coli is dif- cone collapse and turn away of the entire axon (Fig. 2b). ferentially distributed in growth cones and modulates their In contrast, neutralisation of the 20 amino acid repeat steering. J. Neurosci. 27, 12590-12600. domains in the middle region (which are necessary for • Thelen, K., et al. (2007). Cell adhesion molecule DM-GRASP APC’s integration into a degradation complex) leads to the presented as nanopatterns to neurons regulates attachment formation of protrusions and turn of the axons towards this and neurite growth. Soft Matter 3, 1486-1491. side. This shows for the first time the role of APC for a local • Thelen, K., et al. (2008). Ubiquitination and endocytosis of cell and domain specific modulation of microtubule dynamics adhesion molecule DM-GRASP regulate its cell surface presence and affect its role for axon navigation. J. Biol. Chem. 288, in growth cones, thus affecting axonal steering. 32792-32801.

Fig. 2: Role of the microtubule system in growth cones. (a) Laser scanning microscopy visualizes APC and microtubules in the growth cone. (b) Local laser inactivation of the N-terminal region of APC in one growth cone Structure of the Group half leads to a turn of the axon away from this side whereas inactivation of the middle region of APC results in a turn towards this side as seen Group Leader: Gabriele Elisabeth Pollerberg by the growth cone trajectories depicted for the first 10 min after laser Postdoctoral fellows: Christoph Knab, Karsten Thelen treatment (10 axons each). PhD-Students: Jan Hegner, Francisco Ropero, Steffen Jährling, Christian Hahn, Michael Köster, Bettina Maier Undergraduates: Helen Desirée Krause, Friderike Ewald, Fabian Roger, Xiaorui Sun, Jakob Villoth, Technicians: Susanne Bergmann, Claudia Brandel, Monika Zieher-Lorenz

115 Gudrun Rappold Current Research Molecular pathogenesis of genetic disorders Homeodomain Research Summary transcription factors SHOX and SHOX2 The focus of our research is the molecular elucidation of human disease, especially growth and neuronal disor- Homeodomain proteins ders. To uncover the causes of these disorders, our work play a fundamental role employs different cell culture and animal models and dif- during embryogenesis and ferentiated embryonic stem cells. We would like to under- development by regulating stand how mutations correlate with disease, how genes pattern formation and are regulated and how they contribute to differentiation organogenesis. SHOX is a and development. member of the paired-related homeobox family, highly conserved among vertebrate species, but absent in Curriculum Vitae mouse and rat. SHOX is best known as a master controller of human height. Defects in SHOX cause syndromal (Léri- Degrees: 1980 Diploma (M.Sc.) University of Heidelberg 1984 Ph.D. University of Heidelberg Weill, Langer and Turner syndrome) as well as idiopathic 1993 Habilitation growth failure. Together, SHOX defects probably represent 1974-1980: Biology and Chemistry the most common known genetic cause of short stature (Stanford, USA, Konstanz and Heidelberg) in humans. Furthermore, expression in muscle, vascular 1980-1984: Doctoral thesis in Human Genetics, Heidelberg tissue and brain suggests that SHOX also plays a role in 1984-1985: DAAD Scholarship at the Medical Research myogenesis, angiogenesis and neural development. Council, Mammalian Genome Unit, The SHOX gene is regulated both at the transcriptional and Edinburgh, Scotland translational level. To understand the regulation of gene 1985-1987: Postdoctoral fellow at EMBL, Heidelberg expression in a spatial and temporal manner, the study of 1987-1988: Postdoctoral fellow a the Imperial Center Research fund (ICFR) London, England tissue-specific regulatory elements and its consequence 1989: Group Leader at the Institute of Human Genetics, with respect to the associated phenotypes will be carried Heidelberg out in model systems. 2004: Full Professor and Chair of Department of Human The SHOX protein acts as a transcriptional activator which Molecular Genetics, University of Heidelberg binds to specific DNA sequences. Crucial functional domains for SHOX activity (DNA binding, dimerisation, nuc- Contact lear translocation) have been defined and characterised Department of Human Molecular Genetics by mutant variants, providing a mechanistic understan- Institute of Human Genetics ding of the function of this key gene. Phosphorylation on University of Heidelberg Ser 106 has been shown to play an important role in regu- Im Neuenheimer Feld 366 lating SHOX biological activity by modulating its transcrip- 69120 Heidelberg tional activity. Germany SHOX induces cellular growth arrest and is expressed in hu- A +49-6221-565153 man growth plate chondrocytes. It is expressed in human E +49-6221-565155 K [email protected] and chicken embryos from an early stage onwards with o http://www.med.uni-heidelberg.de/Abteilung-Molekulare- high expression in the developing limbs and represents Humangenetik.100279.0.html a marker for chondrocyte maturation in the growth plate. Up to now, there is very little data on the molecular pa-

116 Gudrun Rappold thways of SHOX. Identification of upstream regulators and the following experimental approaches: studies using pri- downstream targets will help to understand the pathways mary chondrocytes and differentiated mesenchymal stem in which this gene functions. cells, Yeast-two-hybrid analysis, GST-pulldown, Co-IP assays, Microarray hybridisation, quantitative RT-PCR, Repor- ter gene assays and animal as well as cell culture models.

Serotonin 5-HT Receptor Genes Serotonin (5-hydroxytryptamine, 5-HT) controls a variety of physiological functions in the central and peripheral nervous system. Serotonin action is mediated by a multitude of 5-HT receptor subtypes divided into seven main classes (5-HT1R to 5-HT7R). Except for the 5-HT3 receptor, which is a ligand-gated ion channel, all serotonin receptors represent G-protein coupled binding proteins. Our previous work comprised the analysis of HTR3A and B genes as candidates in the etiology of psychiatric disorders. In more recent years, we successfully isolated several Fig. 1: SHOX expression domains in developing human limb at CS 18. novel serotonin receptor 5-HT3-genes, which can help unravelling the 5-HT3 receptor diversity in humans. One of On the clinical/translational side, we have established the novel genes (HTR3E) represents an excellent candidate the genotype-phenotype correlation in more than two for the investigation of gastrointestinal diseases such as ir- thousand children with short stature, set up an interactive ritable bowel syndrome and eating disorders (bulimia ner- SHOX database, accompanied a controlled, multi-center vosa and anorexia nervosa) and might shed light upon the trial treating children with SHOX deficiency with recom- pathomechanism of these particular disorders. Analyses binant growth hormone. In 2005, the results of this trial of the HTR3 genes are carried out on an electrophysiolo- were given orphan drug status by the FDA and in 2007, the gical, pharmacological, biochemical as well as cytological treatment on SHOX disorders was approved by the Federal level to investigate the possible molecular make-up of the authorities in the US and Europe. respective receptor subtypes and its response to specific For Shox2, its importance in embryonic development drugs. was recently shown by demonstrating that its complete loss of functions in “knock-out” mouse models is incom- Functional Role of the RhoGAP gene MEGAP in neuronal patible with postnatal life and analyses of Shox2 deficient development embryos revealed highly specific defects within the sinus venosus of the developing heart as the likely cause of em- Mental retardation is a developmental disorder charac- bryonic lethality. Using this mouse model, we are trying terised by a global deficiency in cognitive abilities. It is a to understand the basic functions of Shox2 during early common clinical disorder affecting about 2-3% of the po- organogenesis. Based on the extraordinary high degree pulation. We have found that haploinsufficiency of a Rho- of evolutionary conservation, we are furthermore able to GAP gene, MEGAP, is associated with mental retardation investigate Shox2 functions in diverse animal models in- in several patients. The Rho family of GTPases, and in par- cluding Chicken, Xenopus and Zebrafish. ticular MEGAP, plays an important role in various aspects Our cell and molecular biological analyses include indivi- of neuronal development, including neurite outgrowth dual steps of transcriptional and post-transcriptional regu- and differentiation, axon pathfinding and dendrite spine lation, interacting proteins and downstream targets using formation and maintenance.

117 Gudrun Rappold

Top publications • Muncke, N., et al. (2003). Missense mutations and gene inter- ruption in PROSIT240, a novel TRAP240-like gene, in patients with congenital heart defect. Circulation 108, 2843-2850. • Marchini, A., et al. (2004). The short stature homeodomain protein SHOX induces cellular growth arrest and apoptosis and is expressed in human growth plate chondrocytes. J. Biol. Chem. 279, 37103-37104. • Kirsch, S., et al. (2005). Interchromosomal segmental duplica- tions of the pericentromeric region of the human Y chromosome. Genome Res. 15, 195-204. • Schneider, K.U., et al. (2005). Identification of a Major Recom- bination Hotspot in Patients with Short Stature and SHOX De- ficiency. Am. J. Hum. Genet. 77, 89-96. Fig. 2: Immunostaining of a transverse section of an E 11.5 mouse • Blaschke, R., et al. (2006). The pseudoautosomal regions, SHOX embryo at the thoracic level. Commissural axons were visualized using Tag-1 (green) and cell bodies with DAPI (blue). Megap (red) is expressed and disease. Curr. Opin. Genet. Dev. 16(3), 233-239. throughout the spinal cord, especially in the commissural axon fibers. • Yang, Y., et al. (2006). MEGAP impedes cell migration via regulating actin, microtubule dynamics and focal contacts formation. Exp. Cell Res. 312, 2379-2393. MEGAP, also called srGAP3, is highly expressed in the • Blaschke, R.J., et al. (2007). Targeted mutation reveals essential developing and mature mammalian nervous system. We functions of the homeodomain transcription factor Shox2 in are interested in the question of how mutations in MEGAP sinoatrial and pacemaking development. Circulation 115(14), that alter Rac1 and Cdc42 signalling result in abnormal 1830-1838. neuronal development and deficient cognitive functioning • Marchini, A., et al. (2007). BNP is a transcriptional target of the in human and mouse. We are studying the mechanisms short stature homeobox gene SHOX. Hum. Mol. Genet. 16(24), of MEGAP regulation during development in primary 3081-7. neurons and axonal growth cones with regard to actin and • Sabherwal, N., et al. (2007). Long-range conserved non-coding SHOX sequences regulate expression in developing chicken microtubule dynamics and growth cone guidance. limb and are associated with short stature phenotypes in hu- We also analyse MEGAP binding partners and further man patients. Hum. Mol. Genet. 16(2), 210-222. characterise the role of MEGAP in the Slit-Robo signalling • Kapeller, J., et al. (2008). A functional variant in the miR-510 pathway. To study the normal and pathological role of target site of the serotonin receptor type 3E gen eis associ- MEGAP, a targeted gene disruption of the mouse ortholog ated with diarrhea predominant irritable bowel syndrome. has been carried out (collaboration with Prof. Dusan Hum. Mol. Genet. 17(19), 2967-77. Bartsch). The analysis of the knock-out and wildtype mice including analysis of brain sections during different stages of development, behavioural testings and cultures of primary hippocampal neurons, may enable us to better Structure of the Group understand the pathomechanism leading to mental retardation in patients with MEGAP haploinsufficiency. Group Leader: Gudrun Rappold Postdoctoral fellows: Claire Bacon, Jianjun Chen, Volker Endris, Beate Niesler, Katja Schneider PhD-Students: Eva Decker, Claudia Durand, Anne Glaser, Christian Hammer, Johannes Kapeller, Lydia Haussmann, Sandra Puskaric, Simone Steinbrecht Technicians: Elke Fenner, Ralph Roeth, Birgit Weiß

118 Current Research André Rupp

Neuromagnetic Auditory evoked fields, auditory modelling, representation of temporal and psychoacoustics and spectral pitch in the auditory cortex Research Summary Temporal integration in the The focus of our research is the source analysis auditory system within 30 of auditory evoked fields (AEF) recorded using milliseconds plays a crucial magnetoencephalography (MEG) in human listeners. The role in pitch processing. aim is to bridge the gap between perception as revealed Current models suppose that by psychoacoustics and simulations of spike patterns pitch perception is based on of the auditory nerve to support the development and the processing of (i) the temporal regularity of a sound and to assess the validity of models that account for the (ii) its spectral envelope. Although, the representation and perception of pitch, size, modulation detection, and integration of both characteristics remains unclear, the masking properties. latency of the late AEFs exhibit a high correlation to the inverse of the perceived pitch (Gutschalk et al., 2002; Ritter Curriculum Vitae et al., 2005, 2007; Rupp et al., 2005). In an interdisciplinary Degrees: 1992 Diploma (M.Sc) in Psychology, project (Institut für Theoretische Physik Heidelberg, Prof. University of Heidelberg Dosch, and the Centre for the Neural Basis of Hearing, 1999 Ph.D. University of Heidelberg Department of Physiology in Cambridge, Prof. Patterson) 1992-1996: Staff scientist at the Children University Hospital, we are developing critical stimuli by manipulating the Heidelberg, Head of the data center of the temporal regularity as well as the temporal and spectral German Collaborative Study in Phenylketonuria 1996-1998: Thesis work and Research scientist at the envelope of sounds. Spatio-temporal source analysis will be Department of Pediatric, Neurology, applied to locate pitch-related centers in the audiory cortex University of Heidelberg and to investigate the temporal course of the AEF. These 1998-2000: Research scientist at the Division of experiments will be carried out in subjects who perceive Biomagnetism, Department of Neurology, dominantly the fundamental pitch of an uncomplete University Hospital, Heidelberg harmonic complex sound and a complementary group of since 2000: Head of the Section of Biomagnetism, listeners, who dominantly perceive specific harmonics of Department of Neurology such sounds. The first group exhibits larger AEF and larger grey matter volumes in the left hemisphere while the Contact latter shows the opposite behaviour. The choice of these Department of Neurology subjects will provide valuable information to investigate University Hospital Heidelberg the dependency of pitch perception on temporal and Im Neuenheimer Feld 400 spectral aspects. 69120 Heidelberg Germany A +49-6221-567537 Differences in auditory evoked fields and grey matter E +49-6221-565258 volume of the left and right Heschl’s gyrus between K [email protected] musicians and non-musicians o http://www.klinikum.uni-heidelberg.de/biomag The extensive research on auditory fields and morphological characteristics (Schneider et al., 2002; 2005) showed

119 André Rupp that musicians exhibit enhanced P30 components (lo- long-term duration of professional training while the P30 cated in the primary auditory cortex, i.e. medial Heschl’s amplitude is affected to a much lesser degree. gyrus) and P50-components (located in lateral Heschl’s gyrus) compared to non-musicians. Furthermore, the ma- Early representation of the auditory nerve spike pattern gnitude of these signals are correlated with (i) the absolute in the primary auditory cortex grey matter volume of these areas and (ii) a psychometric test to assess musicality. Currently, these investigations Alternating the phases of a harmonic tone produces will be continued in a cross-sectional and longitudinal de- pure-tone masked thresholds differing by more than 20 sign in children and adolescents to further investigate the dB. This effect is due to the characteristics of the auditory influence of training on the plasticity of these components, filters on the basilar membrane which differently affect because the P50 amplitude is strongly correlated to the ringing within channels. Based on the analysis of early

Fig. 1: Simulations of the neural activity in the auditory nerve of a harmo- determines the salience of pitch. (c) The magnitude of the N100 evoked nic complex sound f0=250 Hz (a, tonotopic axis with high frequencies at by tones with different temporal envelopes (ramped vs. damped with dif- the top and low frequencies at the bottom), and the stabilized auditory ferent half-life-times) as derived from sources in the lateral Heschl’s gyrus image (b) that results from an alignment across channels due to a non- correspond to both, (d) the psychoacoustically determined salience of linear strobing mechanism. This computational stage results in a buffer these tones when compared to each other, and (e) the height of the first with a decay of excitation within 32 ms. The distance between the ridges ridge of the stabilized auditory image. corresponds to the perceived pitch of a tone and the height of the ridges

120 André Rupp

AEFs and basilar membrane simulations (Rupp et al., Top publications 2002), we analyze in a collaboration with Prof. Dau (DTU • Sieroka, N.,et al. (2003). Additional neuromagnetic source acti- Oersted, Denmark) the representation of such peripheral vity outside the auditory cortex in duration discrimination cor- effects in the auditory cortex. First results indicate that relates with behavioural ability. NeuroImage 20, 1697-1703. neuromagnetic responses are highly consistent with • Gutschalk, A., et al. (2004). Temporal dynamics of pitch in hu- perceptual properties obtained with the same stimuli man auditory cortex. NeuroImage 22, 755-766. and with results from simulations of neural activity at • Gutschalk, A., et al. (2004). Recovery and refractoriness of audi- the output of cochlear preprocessing. This suggests that tory evoked fields after gaps in click trains. EJN 20, 3141-3147. the activity wave in the auditory nerve keeps its across- • Rupp, A., et al. (2004). Middle latency evoked fields reflect frequency timing structure travelling to the auditory psychoacoustic gap detection thresholds in listeners. J. Neu- rophysiol. 92, 2239-2247. cortex and that the AEF that is reflected in the MEG • Gutschalk, A., et al. (2005). Neuromagnetic correlates of strea- response is strongly related to the timing structures. Due ming in human auditory cortex. J. Neurosci. 25, 5382-5388. to the high correlation of basilar membrane characteristics • Schneider, P., et al. (2005). Structural and functional asymmet- and the morphology of the auditory evoked fields, we are ry of lateral Heschl’s gyrus reflects pitch perception preference. currently optimizing semi-realistic models of the cochlea Nat. Neurosci. 8, 1241-1247. to enhance the validity of spike simulations in the auditory • Ritter, S., et al. (2005). Neuromagnetic responses reflect psy- nerve (Sieroka et al., 2006). Such investigations might choacoustical findings in temporal pitch changes. NeuroI- support the development of hearing aid algorithms that mage 27, 533-543.Gutschalk, A., et al. (2007). The effect of simulate the compressive behaviour of the cochlea which temporal context on the sustained pitch response in human auditory cortex. Cerebral Cortex 17, 552-561. is found in normal hearing subjects. • Buecker, M., et al. (2008). Parallel minimum p-norm solution of the neuromagnetic inverse problem for realistic signals using exact Hessian-vector products. Soc. Ind. Appl. Math. 30, 2905- 2921. • Rupp, A., et al. (2008). Representation of auditory filter phase characteristics in the cortex of human listeners. J. Neurophy- siol. 99, 1152-1162.

Structure of the Group Group Leader: André Rupp Postdoctoral fellows: Steffen Ritter Undergraduates: Martin Andermann, Johannes Hack Technicians: Barbara Burghardt, Esther Tauberschmidt

121 Martin Schmelz Current Research Translational pain research Cellular porcine model Research Summary Patch-clamp and histoche- In this research group sensitization of primary afferent mistry of porcine dorsal root nociceptors is investigated as one mechanism for chronic ganglion cells to characteri- pain and neuropathy. We focus on translational studies ze different nociceptor clas- using single fiber recording techniques in chronic pain ses. patients, human volunteers and large animals (pig) Development of in vitro mo- combined with functional and structural investigations del of nerve endings separa- of skin innervation in vivo and in cell culture. ted from the cell body and coculture with keratinocytes. Curriculum Vitae Porcine single fiber recordings Degrees: 1991 M.D. University of Erlangen 1999 Habilitation Characterization of different afferent nerve fiber classes 1985-1991: Medical school and modulation of axonal excitability using standard 1991-1992: Internship at the Department of Occupational teased fiber techniques in the pig saphenous nerve. Medicine, University of Erlangen Effects of locally applied growth factors on excitability of 1993-1999: Postdoctoral fellow at the Department of nerve endings and axonal excitability. Physiology, University of Erlangen 1999-2002: Assistant Professor at the Department of Porcine neurogenic inflammation Physiology, University of Erlangen since 2002: Karl-Feuerstein Professor, Department of Development of experimental models of peripheral sen- Anesthesiology Mannheim, sitization by UVB irradiation with the objective readout of University of Heidelberg, axon reflex erythema measured by laser Doppler imaging. Heading the section for Experimental Microdialysis in the sunburn to assess levels of inflamm- Pain Research atory mediators involved in peripheral sensitization. 2004-2007: Speaker of the Clinical Research Group KFO 107 since 2007: IZN Investigator Human pain models Contact Characterization of human pain models for peripheral (UVB Department of Anesthesiology, Mannheim irradiation) and central (electrical pain and hyperalgesia University of Heidelberg model) sensitization by pharmacological intervention. Theodor Kutzer Ufer 1-3 Objective assessment of cutaneous unmyelinated inner- 68167 Mannheim vation by electrically induced axon reflex erythema and Germany electrically induced axon reflex sweating. A +49-621-3835015 E +49-621-3835015 Chronic pain patients K [email protected] o http://www.klinikum-mannheim.de/2288.0.html Microneurographic characterization of neuropathic changes in primary afferent fibers linked to chronic pain. Correlation of functional and structural changes in neuropathic pain by using skin biopsies, quantitative sensory testing, and electrically induced axon reflex measurements.

122 Martin Schmelz

Fig. 1: Two-compartment in vitro model of piglet DRG neurons with somata and processes separated by a diffusion barrier. Microphotographs after 6 days in cell culture. PGP9.5 staining.

Top publications • Orstavik, K., et al. (2003). Pathological C-fibres in patients with a chronic painful condition. Brain 126, 567-578. • Steinhoff, M., et al. (2003). Proteinase-Activated Receptor-2 Mediates Itch: A Novel Pathway for Pruritus in Human Skin. J. Neurosci. 23, 6176-6180. • Yosipovitch, G., et al. (2003). Itch. Lancet 361, 690-694. • Kramer, H.H., et al. (2004). Electrically stimulated axon reflexes are diminished in diabetic small fiber neuropathies. Diabetes 53, 769-774. • Ikoma, A., et al. (2006). The neurobiology of itch. Nat. Rev. Neu- rosci. 7, 535-547. • Orstavik, K., et al. (2006). Abnormal function of C-fibers in patients with diabetic neuropathy. J. Neurosci. 26, 11287-11294.

Fig. 2: Corresponding patterns of activity dependent slowing of conduction velocity (360 stimuli at 2Hz, followed by 40 stimuli at 0.25Hz) in respective C-fiber classes in humans (microneurography) and pig (teased fiber technique). cmi: mechano-insensitive nociceptors, cm: polymodal nociceptors, symp: sympathetic efferent, cold: cold nociceptors.

Structure of the Group Group Leader: Martin Schmelz Senior scientists: Marlen Petersen Postdoctoral fellows: Roman Rukwied Scientists: Martin Dusch, Otilia Obreja, Marcus Schley Technicians: Andreas Klusch, Elmar Forsch

123 Gerhard Schratt Current Research The role of microRNAs in synaptic develop- Genome-wide identifica- ment and plasticity tion of microRNAs that function during synapse Research Summary development The local translation of mRNAs near synapses is crucial MicroRNAs are a class of for various forms of synaptic plasticity. We recently found non-coding RNAs (18- that microRNAs, an extensive class of small non-coding 25 nt) that act as post- RNAs, regulate synaptic protein synthesis and dendritic transcriptional regulators spine development in neurons. Future experiments aim of gene expression. In the at the identification of the full complement of synaptic mammalian system, the microRNAs and their target mRNAs, as well as microRNA majority of microRNAs bind function in memory-related processes in vivo. to 3’UTR regions of target mRNAs, thereby inhibiting Curriculum Vitae the translation of a large set of target mRNAs. In recent years, microRNAs have been attributed a flurry of cellular Degrees: 1998 M.Sc. (Diploma) University of Tübingen functions, including the regulation of cell differentiation, 2002 Ph.D. University of Tübingen survival, and metabolism (Bartel, 2004). The function of 1998-2002: Thesis work, Interdisciplinary Institute of Cell Biology, University of Tübingen microRNAs in the nervous system is still largely elusive. 2002-2006: Postdoctoral fellow at the Division of However, the complete lack of microRNAs in the brain Neuroscience, Chrildren‘s Hospital and leads to defects in morphogenesis and survival. In addition, Department of Neurobiology, examples of individual microRNA functions in neurons Harvard Medical School, Boston, USA have been reported, in particular during early stages of since 2006: IZN Investigator neuronal fate determination (miR-9, miR-124) and neurite outgrowth (miR-132) (Kosik, 2006). Recently, we found that Contact a brain-specific microRNA, miR-134, is expressed in post- SFB488 Junior Group, mitotic neurons and involved in morphological regulation IZN of dendritic spines, the major sites of excitatory synaptic University of Heidelberg transmission (Schratt et al., 2006). This finding led to the Im Neuenheimer Feld 345, 1.OG hypothesis that a microRNA regulatory network could 69120 Heidelberg control the expression of critical synaptic proteins during Germany synaptic development and plasticity. A +49-6221-566210 E +49-6221-567897 In an attempt to characterize the full complement of K [email protected] microRNAs in the synaptodendritic compartment, the o http://izn.uni-hd.de/ synaptic “microRNome”, we embarked on a large-scale expression screen using microarray profiling of microRNAs isolated from rat synaptosome preparations (Siegel et al., 2009). Thereby, we obtained a list of microRNAs that were significantly enriched in synaptosomes compared to whole brain preparations. Subsequent loss-of-function experiments revealed that a subset of these microRNAs is involved in bi-directional regulation of dendritic spine size

124 Gerhard Schratt

(Fig. 1). One of the identified miRNAs, miR-138, appears to Identification of microRNA-associated protein comple- control spine size by regulating the palmitoylation status xes in neurons of critical synaptic proteins. These results will provide a We recently found that the inhibitory activity of miR-134, in roadmap for future experiments to determine the role addition to calcium-dependent transcription, is regulated of individual miRNAs in synaptic plasticity in vivo (see at the post-transcriptional level by the neurotrophin below). BDNF. This regulation likely involves post-transcriptional modification of a miR-134 associated protein/RNA Activity-dependent regulation of microRNA expression complex, since miR-134 binding to its target mRNA Limk1 and function appears not be altered by BDNF (Fig. 2). To get a handle Synapse development and plasticity are highly regulated on the protein components of the complex we currently by neuronal activity. Interestingly, we recently discovered perform a large-scale RNA interference screen, targeting that expression of the synaptic miR-134 is induced by neuronal RNA-binding proteins that might participate in depolarization of the neuronal membrane (Fiore et al., microRNA-dependent gene regulation. This approach will 2009). miR-134 is a member of a large microRNA cluster be complemented by the biochemical characterization that consists of more than 40 miRNAs, and it appears that of the miR-134 associated protein complex and the the entire cluster represents one transcriptional unit that functional characterization of RNA-binding proteins in is co-regulated by a common calcium sensitive regulatory dendritic spine development. element. Currently, we are trying to identify the cis-acting regulatory factor(s) that couple miRNA cluster expression Large-scale identification of physiological microRNA tar- to neuronal activity, and to understand the role of calcium- get mRNAs in neurons induced miRNA expression in activity-dependent processes The functional characterization of microRNAs is still such as dendrite growth, synaptogenesis and plasticity. hampered by the limited knowledge of physiological microRNA target mRNAs. Bioinformatic algorithms have been generated to predict targets based on sequence complementarity and accessibility. These algorithms however do not take into account cell-type specific expression and subcellular localization of microRNAs and their target mRNAs. We plan to exploit the fact that miRNAs primarily regulate mRNA translation at the level of initiation to elucidate microRNA targets. Therefore, polysome- bound mRNAs from neurons will be profiled (Schratt et al., 2004), and those mRNAs that display differential polysome association as a function of microRNA activity will be screened for potential microRNA binding sites.

In vivo functional analysis of neuronal microRNAs Fig. 1: Model for the role of microRNAs in the regulation of G protein signaling and dendritic spine morphogenesis. As previously shown, miR- Although the evidence for a role of microRNAs in 132 and miR-134 antagonistically regulate activity of the spine growth postmitotic neurons is growing, the in vivo function of promoting Rac signaling pathway. In addition, miR-138 might activate individual miRNAs in synaptic development and function the Rho signaling pathway implicated in spine shrinkage by promoting is still completely unknown. We plan to pursue two membrane-association of Galpha. RISC: RNAi-induced silencing complex. ROK: Rho-associated kinase. EphR: Ephrin receptor. APT1: Acyl-protein complementary approaches to unravel the physiological thioesterase 1. Limk: Lim-domain containing protein kinase 1. role of candidate miRNAs that we recently identified in

125 Gerhard Schratt cultured neurons: i) virus-mediated delivery of microRNAs Top publications or their inhibitors into mice using intracranial injection. Ii) • Schratt, G.M., et al. (2004). BDNF regulates the translation of a classical site-directed mutagenesis in the mouse, using select group of mRNAs by a mammalian target of rapamycin- the Cre-loxP system to specifically inactivate microRNAs phosphatidylinositol 3-kinase-dependent pathway during in the postnatal brain. Neurons from virus-infected or neuronal development. J. Neurosci. 24, 9366-9377. genetically modified mice will be subsequently analyzed • Schratt, G. et al. (2004). SRF regulates Bcl-2 expression and at the morphological and electrophysiological level. In promotes cell survival during murine embryonic develop- addition, these mouse models will allow the performance ment. Embo J 23, 1834-1844. of behavioural experiments related to learning and • Schratt, G.M. et al. (2006). A brain-specific microRNA regulates memory. dendritic spine development. Nature 439, 283-289. • Satterlee, J.S., et al. (2007). Noncoding RNAs in the brain. J. Neurosci. 27, 11856-11859. microRNAs as novel targets for therapeutic intervention • Fiore, R., (2007). MicroRNAs in vertebrate synapse develop- Many neurological disorders are characterized by synaptic ment. The Sci. World J. 7, 167-177. dysfunction, including mental retardations, autism- • Fiore, R., et al. (2007). MicroRNAs in synapse development: tiny molecules to remember. Exp. Opin. Biol. Therapy 7, 1823- spectrum and mood disorders. Given our findings from 1831. cultured neurons, we speculate that microRNA function • Fiore, R., et al. (2008). MicroRNA function in neuronal develop- might also be frequently disturbed in these cognitive ment, plasticity and disease. Biochim. Biophys. Acta. diseases. In collaboration with Santaris Pharma, Denmark, • Bicker, S., et al. (2008). microRNAs: tiny regulators of synap- we are currently developing oligonucleotide inhibitors se function in development and disease. J. Cell. Mol. Med 12, directed against synaptic microRNAs for the delivery into 1466-1476. the rodent brain in vivo. In the future, such agents could • Siegel, G. et al. (2009). A functional screen implicates microR- represent a novel avenue for therapeutic intervention. NA-138-dependent regulation of the depalmitoylation enzyme APT1 in dendritic spine morphogenesis. Nat. Cell Biol. 11, 705-716 • Fiore, R. et al. (2009). Mef2-mediated transcription of the miR379-410 cluster regulates activity-dependent dendritoge- nesis by fine-tuning Pumilio2 protein levels. Embo J. 28, 697- 710

Structure of the Group Fig. 2: Activity-dependent regulation of dendritic microRNA complexes. Under basal conditions (left), dendritic microRNAs such as miR-134 Group Leader: Gerhard Schratt recruit miRISC, leading to translational inhibition of dendritic mRNA Postdoctoral fellows: Roberto Fiore targets (i.e. Limk1) and restricted spine growth. Neuronal activity (right) PhD-Students: Guney Akbalik (Erasmus), Silvia Bicker, promotes the release of BDNF, which triggers the activation of the mTOR Mette Christensen (visiting) signaling pathway. This in turn leads to the release of miRISC inhibition Sharof Khudayberdiev, Gabriele Siegel by an unknown mechanism, allowing the translation of dendritic target Undergraduates: Matthias Veith mRNAs and spine growth. Adapted from Schratt et al., 2006. Technicians: Tatjana Wüst

126 Current Research Christoph Schuster Our research targets the Mechanisms of experience-dependent physiological, cellular and synaptic plasticity, learning & memory and molecular mechanisms underlying experience-depen- memory extinction dent synaptic plasticity in Drosophila. Fundamental for Research Summary this research are two com- The physiological, cellular and molecular characterization plementary experimental of synaptic plasticity and higher brain functions such as approaches: learning & memory is often hindered by the high complexity of mammalian brains. We therefore apply powerful genetic, physiological and behavioral tools established in the fruit A) The neuromuscular junc- fly Drosophila melanogaster to study experience-dependent tions (NMJs) of Drosophila larvae represent exceptionally synaptic potentiation of larval glutamatergic synapses with well accessible glutamatergic synapses, which allow in vivo high resolution as well as long-term memory (LTM) formation behavioral stimulation and a highly resolved physiological, and LTM-extinction using an olfactory conditioning paradigm morphological and molecular analysis of the mechanisms in adult flies. Our research aims at facilitating the identification underlying experience-dependent synaptic potentiation. of the principal mechanisms underlying higher brain functions. Curriculum Vitae Degrees: 1988 M.Sc. (Diploma) University of Heidelberg 1992 Ph.D. University of Heidelberg 2002 Habilitation University of Tübingen 1986-1988: University of Heidelberg and EMBL Heidelberg (Biology-Diplom) 1988-1992: Max Planck Institute for Brain Research, Frankfurt (Professor Heinrich Betz) 1993-1996 : Postdoctoral fellow at the University of California at Berkeley, USA 1996-2003: Independent Group Leader at the Friedrich- Miescher-Laboratory of the Max-Planck-Society, Tübingen since 2004: Professor (C3) of Developmental Neurobiology, Interdisciplinary Center for Neurosciences (IZN), University of Heidelberg Contact Fig. 1: Development of larval NMJs of Drosophila. Developing neuro- Department of Neurobiology IZN muscular junction of a Drosophila embryo (A, 12 h after egg laying) and University of Heidelberg larvae (B-D) at different developmental stages (B: 24 h after egg laying; Im Neuenheimer Feld 345 C: 2 days after egg laying, D: 4 days after egg laying). (A) Growth cone 69120 Heidelberg, Germany of the motoneuron aCC extends dorsally towards its target muscle 1. (B) A The growth cone has transformed to presynaptic specialisations of NMJs. +49-6221-548300 These NMJs harbour functionally mature synapses. (C, D) Growth of NMJs E +49-6221-544496 during further larval development. NMJs were labelled with an antibody K [email protected] recognising the cell adhesion molecule Fasciclin II. Scale bars: 10 µm (A), o http://izn.uni-hd.de (B-C); 20 µm (D). (from Schuster et al., 1996a)

127 Christoph Schuster

B) Olfactory fear conditioning of adult flies is a well established paradigm to elicit and analyze various forms of memory (including long-term memory; Fig. 2) and memory modification (such as extinction).

Fig. 2: Dissection of memory phases. At the behavioral level, the observed decay of memory appears relatively seamless (black). Experimental disruptions in numerous animal species including humans, however, reveal temporally, mechanistically and anatomically distinct phases underlying memory retention. In Drosophila, at least four mechanistically distinct phases have been described. These are short-term memory (STM; green), middle-term memory (MTM; blue) anesthesia-resistant memory (ARM; purple) and long-term memory (LTM; red).

Both approaches greatly benefit from the powerful genetic Fig. 3: Phase-I and phase-II of experience-dependent synaptic potentiation. tools available in Drosophila and the possibility to transfer (A) Following the transfer of size-matched food-digging third instar larvae onto a moist, food-free surface, larvae show variable crawling the mechanisms identified in the high-resolution model activities. The crawling profiles of two larvae exhibiting persistently high NMJ to higher brain functions and vice versa. (filled squares) or low crawling activities (open squares) are shown. (B- C) Representative traces of mEJPs (B) and eEJPs (C). (D-E) Time course of Experience-dependent potentiation of synaptic trans- crawling-induced amplitude changes of eEJPs (D) and mEJPs (E) of ‘fast’ and ‘slow’ crawling individuals (filled and open symbols, respectively). The mission presence or absence of changes in the mEJP amplitude and enhanced The crawling activities of Drosophila larvae show large eEJPs define phase-I and phase-II of synaptic potentiation. Data represent mean ± S.E.M. of 9-19 larvae per data point. **p<0.01; *p<0.05. individual differences over time. These differences in crawling profiles are associated with differences in the usage of the glutamatergic neuromuscular synapses and can therefore be used to systematically assess potential characterize the physiological and molecular mechanisms experience-dependent synaptic changes. Based on this underlying experience-dependent synaptic potentiation. strategy we have recently shown that the strength of We are currently focusing on the following topics: glutamatergic transmission can undergo robust and long- lasting potentiation in an experience-dependent manner. • Experience-dependent regulation of presynaptic quantal size This potentiation is mediated by several different synaptic • Role of presynaptic mGluRs in the regulation and mechanisms, which based on their temporal appearance release of large vesicles define a hierarchy of several discrete phases of experience- • Experience-dependent regulation of the functional dependent synaptic potentiation [4]. In this project we balance of postsynaptic GluRs

128 Christoph Schuster

• Experience-dependent regulation of postsynaptic NO- • Characterization of LTM extinction synthase (NOS) activity • Mapping of circuits and neurons involved in LTM • Presynaptic NMDARs and their effect on the extinction probability of vesicle release • Signaling pathways involved in the formation of • Ca2+-dynamics in the pre- and postsynaptic terminal extinction memory during experience-dependent potentiation • Pharmacological and molecular interference with LTM • Role of NOS-activity in the structural organization of extinction synapses • Experience-dependent synaptic protein synthesis and morphological consolidation of synaptic potentiation • Development of a computational model of a simple Top publications network of glutamatergic synapses • Sigrist, S. J., et al. (2000). Postsynaptic translation affects the efficacy and morphology of neuromuscular junctions. Nature Long-term memory formation and LTM extinction in 405, 1062-1065. Drosophila • Sigrist, S. J., et al. (2003). Experience-dependent strengthe- Animals can store information based on their individual ning of Drosophila neuromuscular junctions. J. Neurosci. 23, experiences. Yet not all information is stored for long 6546-6556. • Schuster, C. M. (2006). Experience-dependent potentiation of periods of time. Rather, it is the most intense or even larval neuromuscular synapses. Int. Rev. Neurobiol. 75, 307- traumatic events, or the most repeated information, 322. that is eventually encoded in long-term memory (LTM). • Schuster, C. M. (2006). Glutamatergic synapses of Drosophi- LTM itself and/or access to the stored information is la neuromuscular junctions: a high-resolution model for the continually modified by experience, re-enforced through analysis of experience-dependent potentiation. Cell Tissue reconsolidation or diminished through extinction. Despite Res. 326, 287-299. of an increasing general interest in these phenomena the • Steinert, J. R., et al. (2006). Experience-dependent formation underlying cell biological or molecular mechanisms are and recruitment of large vesicles from reserve pool. Neuron poorly understood. The fruit fly Drosophila has served as 50, 723-733. • Steinert, J. R., et al. (2009). Retrograde Nitric Oxide Signaling a model system for studying various forms of learning Activates Presynaptic NMDA Receptors. Neuron, under revisi- and memory, particularly for those evoked by Pavlovian on. conditioning, largely because of the power of its genetic tools, its smaller central nervous system and fewer molecular redundancies when compared to mammals. Identifying the principal mechanisms underlying LTM formation, consolidation, recall and extinction should facilitate a better understanding of similar processes in mammals and may guide towards novel treatments of psychopathological conditions. Based on established Structure of the Group olfactory fear conditioning paradigms we are currently Group Leader: ChristophSchuster focusing on the following topics: Postdoctoral fellows: Matthias Knirr, Carla Margulies, Jonathan Rojo-Ruiz • Signaling events involved in the formation, PhD-Students: Jean-Louis Thomas, Lucas Vicuña consolidation and recall of LTM Undergraduates: Matthias Heindorf, Jon Leevmann, • Mapping of LTM traces in the fly brain Friedrich Meyer, Roland Svensson • Role of NMDARs in LTM Technicians: Emilia Sancho-Vargas

129 Günther Schütz Current Research Molecular genetics of signal-dependent gene expression I. Nuclear Receptor Function Research Summary Genetic approaches to To understand the role of nuclear receptors, mutations in define the role of the the mouse including cell-specific and inducible mutations estradiol receptor α in have been generated. This approach allowed to define the regulation of the e.g. the role of the glucocorticoid receptor in control of reproductive axis body weight and its function in the brain (Figure 1), of the mineralocorticoid receptor in behavioural plasticity, To define the role of the and established the mechanism of positive feedback of G protein-coupled receptor GPR54 in the estrogen- the estradiol receptor in the brain. The orphan receptor mediated feedback regulation of GnRH synthesis we tailless is crucial for neurogenesis in the adult and is follow two approaches. GPR54 expression is controlled by involved in brain tumor formation (Figure 2). the decapeptide Kiss1 synthesized in the AVPV neurons expressing the estradiol receptor α. Since germ line mutants Curriculum Vitae of GPRR54 are infertile and have a similar phenoptype as Degrees: 1967 MD the ERα forebrain-specific mutants we reason that ERα 1979 Habilitation (Physiological Chemistry) leads to increased synthesis of Kiss1 which binds and Free University of Berlin activates the GPR54 receptor in GnRH neurons. To test this 1966-1967: Thesis work, Institute of Physiological Chemistry, hypothesis we use GPR54-/- mice (obtained from Schering University of Marburg, Plough) in which we express GPR54 under control of the 1967-1969: Internship, Free University of Berlin GnRH gene. We also generate mice with a floxed allele of 1969-1974: Postdoctoral fellow and research associate, the GPR54 gene which we will cross with forebrain-specific Columbia University, New York, USA 1974-1975: Assistant Professor, Cre-expressing mice. With these experiments we hope Columbia University, New York, USA to provide conclusive evidence that the critical positive 1975-1980: Head of an Independent Research Group, feedback action of estrogens upon GnRH neurons are Max-Planck-Institute of Molecular Genetics, Berlin mediated via the Kiss1/GPR54 system and that the action Since 1980: Head of the Division “Molecular Biology of of the receptor is indirect. the Cell I”, German Cancer Research Center, Heidelberg, and Professor, Faculty of Biology, Evaluation of corticosteroid receptor function in the University of Heidelberg brain Contact Corticosteroid hormones regulate a variety of Helmholtz Professorship Molecular Biology of the Cell I developmental, physiological and pathological German Cancer Research Center (DKFZ) processes by interacting with specific receptors, the Im Neuenheimer Feld 581 glucocorticoid and mineralocorticoid receptor. Using 69120 Heidelberg genetic technologies we have generated a panel of tissue- Germany A +49-6221-423411 specific and function-selective mutations of the genes for E +49-6221-423470 these two corticosteroid hormone receptors in the mouse. K [email protected] These mouse models have allowed us to gain important o http://www.dkfz.de/en/molekularbiologie/index.html insights in corticosteroid hormone function in the animal.

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Using these technologies we have been able to investigate the mineralocorticoid receptor, alone or in combination, are steroid hormone receptor function, both in a tissue- or presently investigated to define the role of these receptors cell type-selective manner since germ line mutations for in functions like learning and memory, hypothalamic the glucocorticoid and mineralocorticoid receptor lead feedback control, and the process of addiction. Function- to lethality. We therefore have had a strong interest to selective mutations combined with gene expression develop conditional alleles for these two receptors. For profiling will help to clarify the transcription mechanism example, using hepatocyte-specific mutations in the involved in the action of corticosteroid hormones in vivo mouse we could show that the glucocorticoid receptor in the brain. and Stat5 in hepatocytes are essential for normal postnatal growth. Surprisingly, the glucocorticoid receptor does The nuclear receptor tailless (Tlx) is required for gene- not bind to DNA but its activity is mediated through ration and maintenance of adult neural stem cells and interaction with the Stat5 protein. To identify the function participates in brain tumor formation of the glucocorticoid receptor in allergic skin conditions The tailless (Tlx) gene encodes an orphan nuclear receptor we have analyzed contact hypersensitivity in various which is expressed in the periventricular neurogenic zone specific glucocorticoid receptor mutant mouse strains. during mouse embryonic development. Mutant mice We could establish that macrophages and neutrophils are survive but display specific anatomical defects in the the targets for immune suppression of glucocorticoids in cortex and the limbic system. In particular late developing contact allergy responses. To generate mutations of the structures such as the upper cortical layers and the dendate gluco- and mineralocorticoid receptor in the adult brain gyrus are reduced in size due to depletion of progenitor we have developed mice which express an inducible Cre cells in the subventricular zone. recombinase in neurons of the forebrain (Figure 1). To determine the cell types which express the Tlx gene These mice with inducible mutations of the gluco- as well as we have chosen an approach where a Cre fusion protein is used as a reporter for the Tlx gene. We have shown that the Tlx gene is expressed exclusively in adult neural stem

Fig. 1: Ablation of GR protein expression in neurons of the adult hippocampus upon tamoxifen treatment, Vibratome sections of mice homozygous for a conditional GR allele (GRflox) and heterozygous for Fig. 2: Cre antibody staining of Tlx-CreERT2 in the subventricular zone the inducible CaMKCreERT2 transgene (GRCaMKCreERT2 mice) have been (SVZ) and rostral migratory stream (RMS) of Tlx-CreERT2-expressing mice analyzed for GR protein expression by immunohistochemistry. after tamoxifen induction.

131 Günther Schütz cells, the B cells, but not any more in progenitors (Figure 3). neurons in the Substantia nigra and Ventral Tegmental Area, Cell fate mapping experiments showed that Tlx-expressing depletion of striatal dopamine content and responsiveness cells are multipotent which can differentiate into neurons, to L-DOPA treatment. astrocytes as well as into oligodentrocytes. Using BrdU- The observation that constitutive and inducible mutations incorporation, GFAP and Cre staining we could show that in the TIF-IA gene mimic so closely Parkinson´s disease these cells are able to self-renew. Thus, Tlx is specifically symptoms, stimulated us to ask whether nucleolar expressed by long-term self-renewing neural stem cells. impairment affects mitochondrial activity and induces Tlx-expressing cells also express CD133, a marker of brain oxidative stress. Ablation of TIF-IA in dopaminergic neurons tumor stem cells. Furthermore, we could show that loss leads to disruption of nucleolar organization as reflected by of Tlx leads to upregulation of PTEN in the subventricular altered distribution of the protein nucleophosmin, which zone compatible with its function in control of neural is specifically found in the nucleolus of wild type mice but stem cell proliferation. These findings establish Tlx as an distributed throughout the entire nucleus in the mutants. indicator for and regulator of adult neural stem cells. Further, disruption of nucleolar integrity due to loss of TIF-IA is associated with the deficiency in the activity of cytochrome c oxidase (COX), the terminal enzyme complex of the mitochondrial respiratory chain. We therefore concluded that nucleolar disruption following TIF-IA ablation leads to impairment of mitochondrial functions. As signs of oxidative damage we find a strong increase in the number of dopaminergic neurons positive for markers of nitrosylated proteins, neuroketals, which are products of oxydised membrane fatty acid components and oxydised DNA. We reason that p53 overexpression mediates the effects of nucleolar disruption on mitochondria.

Fig. 3: The Cre recombinase fusion protein is expressed exclusively in B cells expressing GFAP but not in C or A cells expressing EGFR or doublecortin (DCX), resp.

II. A New Model For Parkinson´s Disease

Disruption of nucleolar function leads to compromised mitochondrial activity We have reported that disruption of nucleolar integrity based on the ablation of the gene for the transcription initiator factor TIF-IA results in stabilization of p53 and p53-mediated apoptosis. These findings are extremely interesting since a crucial role for p53 has been debated in several neurodegenerative diseases. Disruption of nucleolar integrity in dopaminergic neurons following ablation of the transcription initiation factor TIF-IA generated mice with the major hallmarks of Parkinson´s disease: progressive and differential loss of dopaminergic

132 Günther Schütz

Top publications • Mantamadiotis, T., et al. (2002). Disruption of CREB function in brain leads to neurodegeneration. Nat. Genet. 31, 47-54. • Tronche, F., et al. (2004). Glucocorticoid receptor function in hepatocytes is essential to promote post-natal body growth. Genes Dev. 18, 492-497. • Berger, S., et al. (2006). Loss of the limbic mineralocorticoid receptor impairs behavioral plasticity. Proc. Natl. Acad. Sci. USA 103, 195-200. • Wintermantel, T., et al. (2006). Definition of estrogen receptor pathway critical for estrogen positive feedback to gonadotro- pin-releasing hormone neurons and fertility. Neuron 52, 271- 280. • Engblom, D., et al. (2007). Direct glucocorticoid receptor-Stat5 interaction in hepatocytes controls body size and maturation- related gene expression. Genes Dev. 21, 1157-1162. • Parlato, R., et al. (2007). Specific ablation of the transcription factor CREB in sympathetic neurons surprisingly protects against developmentally regulated apoptosis. Dev. 134, 1663- 1670. • Tuckermann, J.P., et al. (2007). Macrophages and neutrophils are the targets for immune suppression by glucocorticoids in contact allergy. J. Clin. Invest. 117, 1381-1390. • Engblom, D., et al. (2008). Glutamate receptors on dopamine neurons control the persistence of cocaine-seeking. Neuron 59, 497-508. • Liu, H.-K., et al. (2008). The nuclear receptor tailless is required for neurogenesis in the adult subventricular zone. Genes Dev. 22, 2473-2478. • Parlato, R., et al. (2008). Activation of an endogenous suicide response after perturbation of rRNA synthesis leads to neurodegeneration in mice. J. Neurosci. 28,12759-12764.

Structure of the Group Group Leader: Günther Schütz Postdoctoral fellows: Stefan Berger, Andrii Domanskyi, Gitta Erdmann, Milen Kirilov, Witold Konopka, Grzegorz Kreiner, Hai-Kun Liu, Rosanna Parlato, Jan Rodriguez Parkitna, Caroline Ronzaud, Ying Wang PhD-Students: Daniel Habermehl, Claus Rieker, Martin Novak Technicians: Heike Alter, Tabea Arnsperger, Dagmar Bock, Katharina Sowodniok, Stefanie Stotz, Andrea Takacs

133 Markus Schwaninger Current Research Gene regulation in cerebral ischemia Transcriptional control Research Summary of ischemic neurodegeneration by NF-kB (Oliver Stroke is a common and devastating disorder but still Herrmann, Waleed Barakat, there is no effective therapy for the majority of patients. Melanie Neubert, Sajjad Because delayed mechanisms of ischemic brain damage Muhammad, Gertrud von such as neuroinflammation, apoptosis, and neurogenesis, Wilpert) rely on gene expression, we investigate gene regulation in the context of cerebral ischemia. Genetic and Apoptosis and inflammation pharmacological tools are combined with the long-term are regulated at the aim to improve stroke therapy. transcriptional level. An essential transcriptional regulator is NF-kB. Our previous work has shown that NF-kB is Curriculum Vitae activated in neurons after stroke. Interference with NF-kB Degrees: 1987 MD signalling reduced the infarct size (Schneider et al., 1999; 2000 Ph.D. Herrmann et al., 2005). Future work will define the relevant 1983-1987: Thesis work in the Institute of Pharmacology, target genes of NF-kB and how the NF-kB signalling University of Freiburg cascade is activated in cerebral ischemia. Furthermore, we 1987-1991: Clinical training in Neurology, FU, Berlin will explore pharmacological tools to interfere with NF-kB 1991-1995: Postdoctoral fellow, Department of Molecular signaling in stroke. Pharmacology, University of Göttingen 1996-2007: Attending physician, Department of TWEAK in cerebral ischemia (Marion Schölzke, Sasidhar Neurology, University of Heidelberg; Murikinati, Nadine Gehrig) Group Leader “Molecular Neuropharmacology” TWEAK is a cytokine of the TNF family. It binds to the Since 2007: Professor of Pharmacology, University of membrane receptor Fn14. In cerebral ischemia, there is Heidelberg a marked up regulation of both the ligand TWEAK and the receptor Fn14. Interference with TWEAK signalling Contact reduced the infarct size two days after onset of ischemia Institute for Pharmacology (Potrovita et al., 2004). However, for clinical application University of Heidelberg further characterization of TWEAK signalling in cerebral Im Neuenheimer Feld 366 ischemia is important. 69120 Heidelberg Germany A +49-6221-548691 Neurogenesis in stroke (Marion Schölzke, Ming-Fei Lang, E +49-6221-548549 Lars Werner, Sasidhar Murikinati, Ira Maegele, Christine K [email protected] Stanek) o http://www.medizinische-fakultaet-hd.uni-heidelberg.de/ Stroke triggers adult neurogenesis. New born neurons are Schwaninger.106141.0.html found in brain areas, in which no neurons are born under normal conditions. Preliminary evidence suggests that the close vicinity to endothelial cells in the vascular niche favours neurogenesis. However, the regulation of ischemia- induced neurogenesis is incompletely understood and

134 Markus Schwaninger evidence for the functional relevance of neurogenesis Top publications is only correlative (Schölzke and Schwaninger, 2007). To • Herrmann, O., et al. (2003). Regulation of body temperature explore neurogenesis in cerebral ischemia, we are working and neuroprotection by endogenous interleukin-6 in cerebral on a transgenic approach to target the relevant cell types ischemia. J. Cerebr. Blood F. Met. 23, 406-415 in vivo. • Acalovschi, D., et al. (2003). Multiple levels of regulation of the IL-6 system in stroke. Stroke 34, 1864-1869. • Schneider, A., et al. (2004). Restriction-mediated differential display (RMDD) indentifies pip92 as a pro-apoptotic gene product induced during focal cerebral ischemia. J. Cerebr. Blood F. Met. 24, 224-236. • Schneider, A., et al. (2004). Identification of regulated genes during permanent focal ischemia: characterization of the protein kinase MARKL1. J Neurochem 88, 1114-1126. • Potrovita, I., et al. (2004). Tumor necrosis factor-like weak in- ducer ofapoptosis-induced neurodegeneration. J Neurosci 24, 8237-8244 • Zhang, W., et al. (2005) Neuronal activation of NF-kB contributes to cell death in cerebral ischemia. J. Cerebr. Blood F. Met. 25, 30-40. • Veltkamp, R., et al. (2005). Hyperbaric oxygen reduces blood- brain barrier damage and edema after transient focal cerebral ischemia. Stroke 36, 1679-1683. Fig. 1: ATP binding site of a kinase up-stream of the transcription factor • Herrmann, O., et al. (2005). IKK mediates ischemia-induced NF-kB. neuronal death. Nat. Med. 11: 1322-1329 • Inta, I., et al. (2006). Bim and Noxa are candidates to mediate the deleterious effect of the NF-κB subunit RelA in cerebral ischemia. J. Neurosci. 26:12896-12903. • Muhammad, S., et al. (2008). The HMGB1 receptor RAGE mediates ischemic brain damage. J. Neurosci. 28, 12023-12031

Structure of the Group Group Leader: Markus Schwaninger Postdoctoral fellows: Dirk Ridder, Antje Krenz PhD-Students: Waleed Barakat, Christoph Leib, Ming-Fei Lang, Sasidhar Murikinati, Fig. 2: Expression of the recombinase Cre (red) in endothelial cells of the Melanie Neubert, Lars Werner cerebellum. Green, endothelial cell marker CD31. Blue, nuclear staining Technicians: Nadine Gehrig, Ira Maegele, by DAPI. Christina Stannek

135 Peter H. Seeburg Top publications: • Jensen, V., et al. (2003). A juvenile form of postsynaptic hippo- Fast excitatory campal long-term potentiation in mice deficient for the AMPA neurotransmission, receptor subunit GluR-A. J. Physiol. 553, 843-856. • Kolleker, A., et al. (2003). Glutamatergic plasticity by synaptic synaptic plasticity and delivery of GluR-B long-containing AMPA receptors. Neuron learning 40, 1199-1212. • Hartner, J., et al. (2004). Liver disintegration in the mouse embryo caused by deficiency in the RNA editing enzyme ADAR1. J. Biol. Chem. 279, 4894-4902. • Schulz, T.W., et al. (2004). Actin/alpha-actinin-dependent transport of AMPA receptors in dendritic spines: role of the PDZ-LIM protein RIL. J. Neurosci. 24, 8584-8594 • Krestel, H.E., et al. (2004). A genetic switch for epilepsy in adult Curriculum Vitae mice. J. Neurosci. 24, 10568-10578. • Aronoff, R., et al. (2004). Neuronal toxicity in C. elegans from Degrees: 1972 M.Sc. (Diploma in Biochemistry) glutamate receptor channels with an ‘edited’ Q/R site. J. Neu- 2000 Ph.D. (Genetics) rosci. 24, 8135-8140. 1972-1975: Thesis work: MPI for Virology, • Hambsch, B., et al. (2005). g-Protocadherins, presenilin-media- Tübingen ted release of C-terminal fragment promotes locus expression. 1974-1975: Assistant Lecturer, University of Heidelberg J. Biol. Chem. 280, 15888-15897. 1975-1977: Postdoctoral fellow, Dept. of Biochemistry and • Kuner, R., et al. (2005). Mechanisms of disease: motoneuron Biophysics, University of California, San Francisco, disease aggravated by transgenic expression of a functionally USA modified AMPA receptor subunit. Ann. N.Y. Acad. Sci. 1053, 1977-1978: Adjunct Assistant Professor in Medicine at UCSF 269-286. 1978-1985: Senior Scientist at Genentech, Inc., • Cetin, A., et al. (2006). Stereotaxic gene delivery in the rodent South Francisco brain. Nat. Protoc. 1, 3166-3173 1985-1987: Staff scientist at Genentech, Inc., South San Francis- • Abraham, N.M., et al. (2009). Synaptic inhibition accelerates co, USA odor discrimination in mice. Neuron, submitted. 1987-1995: Full Professor, University of Heidelberg since 1996: Director at the MPI for Medical Research, Heidelberg

Contact Department of Molecular Neurobiology MPI for Medical Research Structure of the Group Jahnstr. 29, 69120 Heidelberg Group Leader: Peter H. Seeburg A +49-6221-486495 Senior scientists: Miyoko Higuchi, Georg Köhr, E +49-6221-486110 Rolf Sprengel K [email protected] Postdoctoral fellows: Alexander Kolleker, Yair Pilpel, o http://www.mpimf-heidelberg.mpg.de Martin Schwarz PhD-Students: Stefan Bonn, Ali Cetin, Florian Freudenberg, Noam Pilpel Technicians: Horst Großkurth, Sabine Grünewald, Joachim Hopisch, Judith Müller, Margarita Pfeffer

136 Current Research Horst Simon Differentiation and survival of mesencephalic Functional characterization of the Engrailed dopaminergic neurons transcription factors Research Summary The Engrailed genes are Degeneration of dopaminergic neurons in the substantia homeobox transcription fac- nigra is the hallmark of Parkinson’s disease. We examine tors which participate in neu- the molecules, which determine the properties of these ronal specification. They are cells, regulate their differentiation, ensure their survival are expressed in mesence- during development, and play a role in their maintenance phalic dopaminergic (mes- during adulthood DA) briefly after they become postmitotic and critically determine their survival. In mouse embryos deficient of Curriculum Vitae the two genes (En1 and En2), mesDA neurons are generated, start to express their neurotransmitter phenotype, but Degrees: 1989 M. Sc. (Diploma in biochemistry) 1993 Ph.D. then disappear (Alberi et al., 2004; Simon et al., 2001). The 1986-1989: Diploma in Biochemistry and Molecular Biology, Engrailed requirement for the survival of these neurons is ETH Zürich, Switzerland gene-dose dependent and cell-autonomous. Interestingly, 1989-1993: Thesis work at King’s College, in one of the genotypes (En1-/+; En2-/-), which is viable University of London, UK and fertile, the nigral DA neurons are gradually and spe- 1993: Postdoctoral fellow at King’s College, cifically lost during the first two postnatal months (Fig.1). University of London, UK The disappearance of the cells leads to diminished storage 1994-1999: Postdoctoral fellow at The Salk Institute for and release of dopamine in the dorsal striatum (caudate Biological studies, La Jolla, USA putamen) and to motor deficiencies which are reminiscent 1999: Leader of the Biofuture group, University of Heidelberg of akinesia and bradykinesia, two cardinal symptoms of Parkinson’s disease (PD) (Sgado et al., 2006). Contact Death of Engrailed-deficient mesDA neurons is caused by Department of Neuroanatomy University of Heidelberg the activation of the mitochondrial pathway of apoptosis, a Im Neuenheimer Feld 307 molecular mechanism, which is shared with in neurotoxin 69120 Heidelberg based models for PD, intoxication with 1-methyl-4-phenyl- Germany 1,2,3,6-tetrahydropyridine (MPTP), rotenone and 6-hydro- A +49-6221-548342 xydopamine (6-OHDA). An over-expression experiment in E +49-6221-545605 a neuronal cell line demonstrated that the Engrailed genes K [email protected] repress p75NTR, a neurotrophin receptor. Several lines of o http://izn.uni-heidelberg.de/ evidence now suggest that elevated p75NTR expression is causal to the death of engrailed deficient mesDA neurons. Direct interference with p75NTR by a function-blocking antibody or by RNA interference using penetratin-coupled oligos rescues the mutant neurons in vitro. Markedly, the cell death induced by P75 is mediated by the suppression of the Erk1/2 MAPK signalling pathway. (Alavian et al. sub-

137 Horst Simon mitted). We currently investigate this issue further in vivo the excitability of the cells. In mesDA neurons, the channel and vitro, by using mutant mice, viruses and pharmacolo- consists of four inner-core forming Kir6.2 molecules and gical approaches. four surrounding Sur1 molecules. We have shown that genetic inactivation of the channel does not change the survival rate of mesDA neurons during the entire lifespan of null mutants for Sur1 and Kir6.2 (Fig. 2). On the contrary, the in vivo loss of function of the channel results in rescue of nigral DA neurons in two mechanistically distinct mouse models of nigral degeneration, intoxication with MPTP and the weaver mouse (Liss et al., 2005). In our own experiments, we have demonstrated that the same can be achieved by employing channel blockers like the sulfony- Fig. 1: Slow progressive loss of nigral DA neurons in mutant mice hete- lurea tolbutamide. Sulfonylureas are widely used to treat rozygous null for En1 and homozygous null for En2 (EnHT). P0 (A, B), P30 diabetes type 2 in humans. We are currently examining HT (C,D) and 3 months old wild-type (A-F) and En (A’-F’) mice. TH immunos- whether these pharmacological findings can be employed taining on the level of SN (A, C, E) and VTA (B,D,F). to rescue nigral DA neurons from degeneration in models The role of K-ATP channel activity for the sensitivity of for PD. For this purpose, we use Alzet mini-pumps to chro- mesencephalic dopaminergic to mitochondrial insult nically administer K-ATP channel blocking compounds in mouse and rat. K-ATP channels couple the metabolic state of the neuron to its membrane potential by sensing the ATP/ADP ratio. The channel opens, when the intracellular ATP concentration decreases, leading to an outflux of potassium ions and to hyperpolarization of the membrane, thus reducing

Fig. 2: K-ATP channel component Kir6.2 and Sur1. (A) Scheme of K-ATP channel. (B) Cell count of 18 months old wild type and null mutants for Kir6.2 and Sur1. (C) Expression of tyrosine hydroxylase (TH), Sur1 and Kir6.2 in nigral DA neurons.

138 Horst Simon

Top publications • Alberi, L., et al. (2004). Engrailed genes are cell-autonomously required to prevent apoptosis in mesencephalic dopaminergic neurons. Dev. 131, 3229-36. • Thuret, S., et al. (2004). Identification and developmental analysis of genes expressed by dopaminergic neurons of the substantia nigra pars compacta. Mol. Cell. Neurosci. 25, 394-405. • Thuret, S., et al. (2004). The neuregulin receptor, ErbB4, is not required for normal development and adult maintenance of the substantia nigra pars compacta. J. Neurochem. 91, 1302- 11. • Simon, H.H., et al. (2005). Engrailed genes control developmental fate of serotonergic and noradrenergic neurons in mid- and hindbrain in a gene dose-dependent manner. Mol. Cell. Neurosci. 28, 96-105. • Schober, A., et al. (2006). GDNF applied to the MPTP-lesioned nigrostriatal system requires TGF-beta for its neuroprotective action. Neurobiol. Dis. • Scholz, C., et al. (2006). One step cloning of defined DNA fragments from large genomic clones. J. Biochem. Mol. Biol. 39, 464-7. • Sgado, P., et al. (2006). Slow progressive degeneration of nigral dopaminergic neurons in postnatal Engrailed mutant mice. Proc. Natl. Acad. Sci. USA 103, 15242-7. • Gherbassi, D., et al. (2007). Merging mouse transcriptome analyses with Parkinson’s disease linkage studies. DNA Res. 14, 79-89. • Alavian, K.N. (2009). Engrailed Expression in Midbrain Dopa- minergic Neurons Controls Mitochondrial Stability and Erk1/2 Signaling by Regulation of P75/Ngfr. Submitted to Current Biology.

Structure of the Group Group Leader: Horst Simon PhD-Students: Kambiz N. Alavian, Lavinia Alberi, Lavinia Bhatt, Daniel Gherbassi, Christian Scholz, Paola Sgado, Sandrine Thuret Undergraduates: Hima Chandra Technicians: Gabi Döderlein

139 Thomas Söllner Current Research

Molecular mechanisms mediating regulated Neurotransmitter release is exocytosis the paradigm for regulated exocytosis and distinct Research Summary components acting at Intracellular membrane fusion involves the assembly different steps have been of cognate v-SNAREs and t-SNAREs between opposing identified (see Figure 1 for a membranes. At the neuronal synapse, a cascade of com- brief overview). Membrane ponents and reactions controls SNARE complex formation fusion is initiated by the and couples the triggering signal to the fusion machinery. interaction of the v-SNARE To elucidate the molecular function of the regulatory ma- VAMP2 on synaptic vesicles chinery, we are attempting to reconstitute neurotrans- with its cognate t-SNARE, consisting of syntaxin1 and mitter release in vitro. SNAP25 on the presynaptic plasma membrane. SNARE proteins assemble into a stable four-helix bundle. In vitro Curriculum Vitae assays have shown that this protein folding reaction drives Degrees: 1986 M. Sc. (Diploma in Biology) membrane fusion. However, in vivo, SNARE complex 1991 Ph.D. assembly is tightly regulated and involves multiple 1982-1986: Diploma in Biology, steps and numerous regulatory components that either Ludwig-Maximilians University Munich accelerate or arrest the formation of distinct reaction 1987-1991: Thesis work at the Institute for Physiological Chemistry, Biochemistry and Cell Biology, intermediates. The initial tethering of synaptic vesicles to Ludwig-Maximilians-University Munich the active zone of the neuronal synapse does not depend 1991-1993: Research fellow, Cellular Biochemistry and on v-/t-SNARE interactions, but likely requires small GTP- Biophysics, Sloan-Kettering Institute, binding proteins such as Rab3 and its effectors. Further, New York, USA the t-SNARE component syntaxin1 and its binding partner 1994-1997: Assistant Laboratory Member, Cellular Munc18-1 have been implicated in vesicle docking. Thus, Biochemistry and Biophysics, the vesicle tethering machinery is directly linked to the Sloan-Kettering Institute, New York, USA downstream fusion machinery and a multi-step, multi- 1998-2004: Assistant Professor, Cellular Biochemistry and component process controls SNARE complex assembly. Biophysics and Cell Biology programs, Sloan-Kettering Institute, New York, USA The primed pool of synaptic vesicles seems to be docked 2004-2005: Associate Professor, Cell Biology, by partially assembled SNARE complexes and might be Sloan-Kettering Institute, New York, USA arrested at a hemifusion step. Calcium binding to the since 2005: Full Professor at the Biochemistry Center, calcium sensing machinery, which is coupled to the University of Heidelberg SNAREs, opens the fusion pore. Fusion pores have two Contact fates: they can reversibly close or fully dilate resulting in the complete incorporation of the vesicular components Biochemistry Center (BZH) into the plasma membrane. SNAREs are subsequently University of Heidelberg recycled by an energy consuming reaction, involving Im Neuenheimer Feld 345 69120 Heidelberg, Germany the hexameric ATPase NSF. Endocytosis regenerates the A +49-6221-545342 synaptic vesicles. E +49-6221-545341 K [email protected] To analyze the dynamic assembly of the neuronal exocytosis o http://www.uni-heidelberg.de/zentral/bzh/ machinery and to dissect the distinct molecular mecha-

140 Thomas Söllner nisms, we are using completely and semi-reconstituted for efficient endocytosis. fusion assays (Figure 2). These assays allow us to monitor Role of regulatory proteins (Rim1, Rab 3, Munc18, content and lipid mixing and to detect hemifusion Munc13, complexins, synaptotagmins, Vo-ATPase, and intermediates. In addition, we measure regulated other regulators) exocytosis in vivo by using a pH-sensitive GFP targeted Numerous regulators directly bind SNARE proteins, but to secretory vesicles. Cryo-electron microscopy will be in many cases their molecular functions are unclear and employed to analyze the architecture of reconstituted the regulatory network is poorly understood. We study fusion pores. these proteins as individual components and in various combinations. Therefore, these regulators are expressed as recombinant proteins in E.coli, or insect cells, purified to homogeneity, and analyzed in in vitro fusion assays. In biochemical assays, we measure the fusion of an entire population of reconstituted SNARE-liposomes (Figure 2A). In

Fig. 1: Model showing steps and components involved in neuronal exocytosis. Fig. 2: In vitro and in vivo assays. (A) Biochemical fusion assay. v- and t-SNAREs are reconstituted into Specific projects: small unilamellar donor and acceptor liposomes. Role of lipids and membrane microdomains in exo- and Instead of acceptor endocytosis liposomes, synaptic vesicles can be used. t-SNAREs have been found in membrane microdomains Membrane fusion of labeled donor liposomes and a perturbation of these microdomains inhibits with unlabeled exocytosis. Furthermore, the local lipid environment at acceptor liposomes the fusion site could significantly affect the recruitment results in fluorescence of regulatory components, the membrane curvature, and dequenching. (B) Single vesicle docking/fusion assay. A giant unilamellar vesicle, fusion pore dynamics. Indeed recent results indicate that containing v-SNAREs is labeled with DiO-C18 (left panel). Addition of a local lipid turnover is required for exocytosis. Therefore small unilamellar v-SNARE liposomes labeled with rhodamine PE shows we are analyzing the role of protein-lipid interactions vesicle docking (right panel). Scale bar 10 μM. and distinct lipids at different steps of the fusion reaction. (C) In vivo exocytosis assay. Fluorescence surface plots of a PC12 cell transfected with pHluorin targeted to secretory vesicles are shown. Furthermore we will test, whether the unique lipid Membrane depolarization results in a transient increase of pHluorin composition of synaptic vesicles facilitates the sorting of fluorescence at single vesicle fusion sites (compare left panel with 2 right synaptic vesicle components and thus provides a platform panels, s = seconds).

141 Thomas Söllner imaging assays, the docking, hemifusion, and full fusion of Top publications individual SNARE-liposomes can be resolved (Figure 2B). In • Hu, C., et al. (2003). Fusion of cells by flipped SNAREs. Science in vivo assays, the exocytosis of individual secretory vesic- 300, 1745-1749. les is studied in living cells (Figure 2C). In this method, the • Burri, L., et al. (2003). A SNARE required for retrograde trans- secretory granules contain a pH-sensitive marker (pHluo- port to the endoplasmic reticulum. Proc. Natl. Acad. Sci. USA rin) in their lumen. Upon triggering exocytosis, fusion pore 100, 9873-9877. opening and content release is detected by an increase in • Varlamov, O., et al. (2004). i-SNAREs, inhibitory SNAREs that fluorescence. Using siRNA technology, proteins involved fine-tune the specificity of membrane fusion. J. Cell Biol. 164, in exocytosis can be down regulated and replaced by mu- 79-88. tant copies. Thus, the biochemical results obtained in the • Fix, M., et al. (2004). Imaging single membrane fusion events mediated by SNARE proteins. Proc. Natl. Acad. Sci. USA 101, reconstituted assays can be directly tested and verified 7311-7316. under physiological conditions. These combined analyses • Fukasawa, M., et al. (2004). Localization and activity of the shall reveal the sequential order and the entire assembly SNARE Ykt6 determined by its regulatory domain and palmi- line that generates “ready to fuse” vesicles. toylation. Proc. Natl. Acad. Sci. USA 101, 4815-1820. • Söllner, T.H. (2004). Intracellular and viral membrane fusion: a Structural and functional organization of fusion pores uniting mechanism. Curr. Opin. Cell Biol.16, 429-435. • Cheng, Y., et al. (2004). Crystallographic identification of Ca2+ At the electrophysiological level, fusion pores are well and Sr2+ coordination sites in synaptotagmin I C2B domain. defined, but their structural organization is unclear. The Protein Sci. 13, 2665-2672. factors that determine reversible fusion pore opening/ • Cosson, P., et al. (2005). Dynamic transport of SNARE proteins closure or full dilation are largely unknown. Thus we are in the Golgi apparatus. Proc. Natl. Acad. Sci. USA 102, 14647- attempting structural analyses of reconstituted fusion 52. intermediates by cryo-electronmicroscopy. • Söllner, T.H. (2007). Lipid droplets highjack SNAREs. Nat. Cell Biol. 9, 1219-20. • Malsam, J., et al. (2008). Membrane fusion: SNAREs and regulation. Cell. Mol. Life. Sci. 65, 2814-2832.

Structure of the Group Group Leader: Thomas Söllner Postdoctoral fellows: Jörg Malsam PhD-Students: Susanne Kreye, Simone Paulsen, Patricia Rusu, Yvette Schollmeier, Florian Seiler Technicians: Jean Michel Krause

142 Current Research Rainer Spanagel We are using newly estab- The neurobiology of drug addiction lished models to generate alcohol/or drug-dependent Research Summary rats or mice. Behavioural, In the Department of Psychopharmacology our major neurochemical and molecu- interest is in drug abuse research. In particular, we are lar examinations on alcohol working on alcohol dependence and the co-morbidity and drug dependent ro- to anxiety and depression. We have also several projects dents will help us to under- on cocaine, opioids, cannabinoids and nicotine. Our close stand the neurobiological collaboration with the Clinic of Addictive Behaviour at mechanisms of addictive the CIMH (Prof. K. Mann) enables us to rapidly translate behaviour. Studies with mu- our preclinical findings into alcohol/drug dependent tant mice (also derived from an ENU screen) and rats will patients. A further translational arm concerns human help us to identify genes involved in the initiation of drug- genetic studies in combination with neuroimaging. seeking behaviour. In addition, an organbank with different alcohol-preferring rat lines has been established. The Curriculum Vitae aim of the organ bank is the characterisation of molecular cascades involved in alcohol-derived diseases. In a com- Degrees: 1991 Ph.D. 1997 Habilitation parative approach an organbank from alcohol dependent 1989-1991: Thesis work patients is used. Using massive microarray approaches and 1991-1995: Scientific assistant, MPI of Psychiatry, Munich proteomic analysis from the material deriving from this or- 1996-1999: Head of research group: “Drug addiction”, MPI of ganbank, we will be able to pin down molecular networks Psychiatry, Munich that trigger the pathological condition. 1997: Habilitation in Pharmacology and Toxicology, University of Munich 1999: Professor for Psychopharmacology, University of Heidelberg/ Central Institute of Mental Health (CIMH) Mannheim Since 2000: Head of the Department of Psychopharmacology at the CIMH

Contact Department of Psychopharmacology Central Institute of Mental Health J5 68159 Mannheim Germany A +49-621-17036251 E +49-621-17036255 K [email protected] o http://www.zi-mannheim.de

Fig. 1: We are in the course of establishing a central animal and human organbank from alcohol dependent individuals for conducting genomics, transcriptomics and proteomics in liver, pancreas and brain tissue.

143 Rainer Spanagel

The primary goals of our investigations are (i) the identification of risk alleles in the development of addictive behaviour, (ii) the characterisation of neuronal networks mediating drug-taking behaviour (mainly by ph-MRI methods), and (iii) the development of new anti- relapse compounds. Linking genetic and pharmacological findings we finally aim for an individually adapted pharmacotherapy. In doing this, alcohol-dependent animals will be separated into different behavioural and neurobiological phenotypes (e.g. by the use of animal MRI) and will then be treated with a corresponding anti-relapse compound. Treatment responses will finally be correlated with specific genotypes and this pharmacogenetic information will then be translated into alcohol dependent patients.

Fig. 2: The interplay of drugs of abuse and clock genes. A dual role is de- In comparison, we are studying neuroplastic changes scribed, in which clock gene activity determines the efficacy of psychoac- within the reward pathway induced by psychostimulants, tive drugs and in which psychoactive drugs influence clock genes. Thus, opioids, nicotine, and cannabinoids and relate these the expression of these genes modulates the neurochemical state in the mesolimbic system and other brain areas, thereby facilitating or redu- changes to the observed behaviours such as behavioural cing the effects of drugs of abuse or antidepressants. However, these and sensitisation, conditioned place preference, intravenous other psychoactive drugs also influence the activity of clock genes. Once self-administration and reinstatement of drug-seeking the activity of clock genes is altered (red), the neurochemical pathways behaviour. that are modulated by these genes are subsequently affected as well as the physiological and behavioural functions driven by these pathways. Drugs of abuse induce neuroadaptations and thereby alter these biolo- Currently, we are running 12 different research projects gical outputs both directly and indirectly via their action on clock genes in DFG, BMBF and EU-funded networks. In the following (green). section one project is described in more details. a sensitised behavioural response did not occur in Per1 One research project focuses on the involvement of clock mutant mice. In contrast, Per2 mutant mice exhibited genes in the modulation of drug-induced behaviours. A a hyper-sensitised response to cocaine. Conditioned link between the neurobehavioural effects of drugs of place preference experiments revealed similar results: abuse and period gene activity has been first established in Per1 mutant mice showed a complete lack of cocaine Drosophila. Flies show behavioural sensitisation following reward, whereas Per2 mutants displayed a strong cocaine- repeated cocaine administration - a phenomenon that induced place preference. The role of the Per1 gene in the has been implicated in drug craving. Thus, they exposed development of cocaine sensitisation has been confirmed the flies to volatilized free-base cocaine which produces in successive studies conducted in different rodents. a set of behaviours similar to those observed in rodents, Currently, these mice are tested for intravenous cocaine including grooming and enhanced motor activity. Flies self-administration and reinstatement behaviour. In vivo mutant in the period gene did not behaviourally sensitise microdialysis is performed in parallel in order to correlate after repeated exposure to cocaine whereas wild-type extracellular dopamine and glutamate levels in the reward flies showed a strong sensitised response. This finding pathway with the observed behaviours. was further supported in mutant mice lacking functional Per genes. Thus, following repeated cocaine injections, Per1 and Per2 mutant mice have now also been studied

144 Rainer Spanagel in alcohol self-administration experiments. Using operant acamprosate acts mainly on a hyper-glutamatergic state, conditions, Per1 and wild type mice were trained to self- yet having only little effect on a “normal” glutamatergic administer alcohol. Furthermore, extinction sessions state. Therefore, acamprosate should be more effective were introduced, followed by reinstatement measures in reducing alcohol consumption in Per2 mutant than in of ethanol-seeking behavior. In another set of animals, wild type mice. Indeed, following repeated acamprosate the mice were exposed to voluntary long-term alcohol treatment, mutant mice showed decreased alcohol consumption, ensued by a two-month deprivation phase, consumption along with a normalization of extracellular after which the alcohol deprivation effect - which is used glutamate levels in the nucleus accumbens. as a measure of relapse - was examined. Mutant mice did not display a significantly divergent number of reinforced lever presses than wild type animals.

Furthermore, no significant differences between groups were obtained regarding reinstatement of ethanol-seeking behavior. Similar results were obtained in the two bottle free choice paradigm. Specifically, no genotype differences concerning consumption and preference were observed over a broad range of different ethanol concentrations. Moreover, after the deprivation phase, both groups exhibited significant alcohol deprivation effects, yet no genotype differences. These data do not suggest a relationship between the circadian clock gene Per1 and ethanol reinforcement, -seeking and relapse behavior. In contrast, compared to wild type animals, Per2 mutant mice exhibit an enhanced alcohol intake and preference when pharmacologically relevant concentrations are offered.

Alterations in the brain reinforcement system of these mutant mice might drive an enhanced incentive Fig. 3: Mode of action of acamprosate. motivation to consume more alcohol than control animals. Acamprosate exerts its actions through glutamatergic mechanisms. It in- The mesolimbic reinforcement system is modulated by teracts with NMDA and possibly with mGlu5 receptors. Although several various glutamatergic input pathways and in a series of acamprosate binding sites on the NMDA receptor have been examined, experiments it was found that Per2 mutant mice have the exact mode of action on the receptor level remains unclear. However, convincing evidence has been obtained on the neurochemical level [52]. a hyper-glutamatergic state, especially in the nucleus Thus, acamprosate is able to restore the imbalance between excitatory accumbens. Regarding the large evidence given in the and inhibition neurotransmission following chronic alcohol exposure. In literature for an involvement of enhanced glutamate levels particular, acomprosate reduces a hyper-glutamatergic state and should or alterations of the glutamatergic system in excessive therefore be called as a ”anti-glutamatergic“ compound. alcohol consumption, one would expect a massive impact of the Per2 gene mutation on alcohol consumption via These new findings provide a clear link of the mouse Per2 alterations within the glutamatergic system. This idea gene, the glutamatergic system, and excessive alcohol has been further confirmed by examining the effects consumption. However, future animal research ought of acamprosate in these mice. Acamprosate is used in to address the question of whether the Per2 gene, and the clinic for relapse prevention and it is suggested that other clock genes, are also directly implicated in alcohol

145 Rainer Spanagel sensitivity, tolerance, withdrawal and most importantly Top publications in alcohol relapse behaviour. Most importantly, however, • Sillaber, I., et al. (2002). Enhanced and delayed stress-induced the link between Per2 and excessive alcohol consumption alcohol drinking in mice lacking functional CRH1 receptors. in mice could already been translated to humans. Thus, Science 296, 931-33. association studies in different samples have demonstrated • Abarca, C., et al. (2002). Cocaine sensitization and reward are that specific genetic variations of the human PER2 gene influenced by circadian genes and rhythm. Proc. Natl. Acad. are associated with high alcohol consumption. Sci. USA 99, 9026-30. • Spanagel, R., et al. (2002). The neuronal nitric oxide synthase (nNOS) gene is critically involved in neurobehavioral effects of alcohol. J. Neurosci. 22, 8676-83. • Spanagel, R., et al. (2005). The circadian clock gene Period2 alters the glutamatergic system and thereby modulates alcohol consumption. Nat. Med. 11, 35-42. • Sanchis-Segura, C., et al. (2006). Involvement of AMPA receptor GluR-C subunit in alcohol-seeking and relapse. J. Neurosci. 26, 1231-1238. • Spanagel, R., et al. (2008). Drugs for Relapse Prevention of Alcoholism – 10 Years of Progress. Trends Pharmacol. Sci. 29, 109-115. • Schumann, G., et al. (2008). Systematic analysis of glutamatergic neurotransmission genes in alcohol dependence and adolescent risk drinking behaviour. Arch Gen. Psychiat. 65, 826-838. • Wiens, F., et al. (2008). The missing drink: the bertam palm of West-Malaysis gets wild mammals to chronically consume alcohol. Proc. Natl. Acad. Sci. USA 105, 10426-31. • Engblom, D., et al. (2008). Glutamate receptors on dopaminergic neurons control the persistence of drug-seeking. Neuron 59, 497-508. • Bilbao, A., et al. (2008). Loss of the Ca2+/calmodulin-dependent protein kinase type IV in dopaminoceptive neurons enhances behavioral effects of cocaine. Proc. Natl. Acad. Sci. USA 105, 17549-54.

Structure of the Group Group leader: Rainer Spanagel Senior scientists: Peter Gebicke-Haerter, Cornelius Pawlak, Wolfgang Sommer Postdoctoral fellows: Ainhoa Bilbao, Anita Hansson, Anna Molander, Stephanie Perreau-Lenz, Miriam Schneider, Valentina Vengeliene PhD-Students: Chris Maria Feichtinger, Briac Halbout, Fernando Leonardi-Essmann, Nina Reinmuth, Peggy Schneider Technicians: Sabrina Koch, Katja Lankisch, Elisabeth Röbel, Claudia Schäfer

146 Current Research Hartwig Spors

Correlation between odor Spatio-temporal patterns of neuronal activity discrimination perfor- in the healthy and diseased brain mance of mice and odor representation in the olfac- Research Summary tory bulb We examine spatio-temporal stimulus representations in vivo. Using a combination of behavioral tests, electro-, and Odors can be discriminated optophysiological techniques we study how spatial and temporal by mice within a few aspects of these stimulus representations drive downstream hundred milliseconds. Using neurons and finally predict behavior. A genetic model for human an automated olfactometer epilepsy allows us to extend these measurements to determine the discrimination time (DT) changes of population activity in the diseased brain. can be measured for various odor pairs. We compare the Curriculum Vitae DT with the physiological representations of odors on the Degrees: 1999 Staatsexamen Medizin level of the olfactory bulb (OB), as measured using in vivo (National board exam) intrinsic optical signal imaging, calcium dye imaging, and 2002 Ph.D. MPI for medical Research & voltage sensitive dye imaging. We search for predictors of Freiburg University behavioral performance within the measured neuronal 1991-1999: Medical school, Albert Ludwig University, activity. Manipulations that change behavioral and Freiburg 1994-1995: Language and Chinese Medicine, physiological responses (e.g. odor adaptation) are used Nanjing University, China to further test correlations between representations / 1996-1997: Medical student, School of Medicine, processing on one side and behaviour on the other side. Leeds University UK (Fig. 1). (Cooperation with Nixon Abraham, Thomas Kuner, 1996: US Medical Licensing Examination (USMLE), step 1 Andreas Schaefer) 1998: US Medical Licensing Examination (USMLE), step 2 1999-2000: Postdoctoral fellow, Department of Neurobiology, • Spors,H., et al. (2002). Spatio-temporal dynamics of Weizmann Institute of Science, Rehovot Israel odor representations in the Mammalian olfactory bulb. 2000-2002: PhD student, MPI for Medical Research, Heidelberg Neuron 34, 301-315. Department of Cell Physiology Abraham,N.M., et al. (2004). Maintaining accuracy at 2002-2003: Postdoctoral fellow, MPI for Medical Research, Heidel- • berg the expense of speed; stimulus similarity defines odor Department of Cell Physiology discrimination time in mice. Neuron 44, 865-876. 2003-2007 Member of an independent junior research group • Spors,H., et al. (2006). Temporal dynamics and latency funded by the State of Baden-Württemberg (since patterns of receptor neuron input to the olfactory bulb. 2005 Leader of this group), MPI for Medical Research Heidelberg J. Neurosci. 26, 1247-1259. Since 2007: Group Leader, MPI of Biophysics, Department of Molecular Neurogenetics, Frankfurt/M. Spike triggered averaging of spontaneous electrical Contact activity in the neo-cortex Department of Cell Physiology MPI for Medical Research In order to understand cortical information processing Jahnstr. 29, 69120 Heidelberg, Germany it is crucial to understand the functional integration of A +49-6221-486621 individual neurons into the cortical network. Voltage E +49-6221-486459 sensitive dye imaging with high spatial and temporal K [email protected] resolution in the neocortex emphasizes mainly sub- o http://www.mpimf-heidelberg.mpg.de/~spors

147 Hartwig Spors

with action potentials of individual neurons and what aspects of the spatio-temporal patterns are predictive of action potential firing of the individual neurons. (Fig. 2). (cooperation with Thomas Hahn) • Hahn, T., et al. (2006). Individual neurons in the same cortical column are driven by different neuronal subgroups. Soc. Neuroscience Abstr.

Fig. 1: Olfactory bulb activation patterns predict odor discrimination times. (A) Examples of activation patterns measured on the dorsal surface of the olfactory bulb for 4 odor pairs: (+) Carvon vs. (-) Carvon (C+/C-); (+) Octanol vs. (-) Octanol (O+/O-); Eugenol vs. Cineol (Eu/Cin); Ethylbutyrate vs. Amylacetate (EB/AA). (B) Pattern difference (Euclidean distance) of the activation patterns of the odor pairs for two odor concentrations (100 ml/ min or 25 ml/min odorized air) as function of the discrimination time. The difference values are significantly anti-correlated with the discrimination time (r² > 0.9, p<0.05). Fig. 2: Spike triggered patterns of electrical sub-threshold activity differ for two neighboring neurons. Top: Sub threshold population activity, measured with the voltage sensitive dye RH1691, around (± 12. 5 ms) threshold electrical activity of layer 2/3. The combination spontaneous APs of two different neighboring neurons was averaged. with tetrode recordings in layer 2/3 allows us to correlate The minimum and maximum color scale values for each image are given the optically measured input to a neuronal population over in per mille on the top left in each image. Scale bar 1 mm. The position of a wide cortical region (e.g. 4 x 4 mm) with action potentials the tetrode (star in the center) and of the whisker barrels were overlaid post hoc. Bottom: The similarity across frames triggered on APs of the of several neighboring neurons. We ask if specific patterns same neuron (black) is higher than the similarity of frames triggered on and temporal sequences of population activity correlate all APs (red) ignoring the neuron identity.

148 Hartwig Spors

In vivo characterization of a mouse model carrying a point mutation (G2R43Q) that causes absence epilepsy in humans While animal models for epilepsy are abundant, only recently genetic mouse models became available that carry mutations which cause epilepsy in humans. Mice with a point mutation in the Gamma2 subunit of the GABA-A receptor display absence seizures and lowered seizure threshold as do their human counterparts. We set out to determine changes on the network level in vivo measuring spontaneous and evoked population activity before and after challenging the animals with a low dose of the epileptogenic drug pentylentetrazol. Cortical opto- and electrophysiological measurements are combined with targeted single unit recordings in the thalamus in order to localize network changes to the cortex and / or the thalamus. (Fig. 3). (Cooperation with Steven Petrou, University of Melbourne) • Witsch, J.J., et al. (2007). In vivo recordings in a mouse model of a human genetic epilepsy. Abstr. German Neuroscience Meeting Göttingen.

Top publications • Abraham, N.M., et al. (2004). Maintaining accuracy at the ex- Fig. 3: Pentylentetrazol (PTZ) enhances whisker-evoked cortical responses in G2R43Q mice barrel cortex. (A) Voltage sensitive dye imaging signal pense of speed; stimulus similarity defines odor discriminati- amplitude increases after PTZ administration in G2R43Q (note different on time in mice. Neuron 44, 865-876. color bars). Average of 32 stimulus responses. (B) Time course of response • de Kock, C.P., et al. (2007). Layer and cell type specific supra- amplitudes pooled over all animals (G2R43Q (+/d) n=8; wild type (WT) threshold stimulus representation in primary somatosensory n=12) normalized to values before PTZ. cortex. Lond. J. Physiol. • Reidl, J., et al. (2007). Independent component analysis of high-resolution imaging data identifies distinct functional domains. Neuroimage 34, 94-108. • Schaefer, A.T., et al. (2006). Neuronal Oscillations Enhance Sti- Structure of the Group mulus Discrimination by Ensuring Action Potential Precision. Plos Biol. 4(6), e163. Group Leader: Hartwig Spors • Spors, H., et al. (2006). Temporal dynamics and latency pat- Postdoctoral fellows: Frederic von Wegner terns of receptor neuron input to the olfactory bulb. J. Neu- Undergraduates: Rebecca Böhme, Nilufar Shahshahani, rosci. 26, 1247-1259. Dmitrij Turaev • Spors, H. et al. (2007). Male Behavior by Knockout. Neuron 55, MD students: Daniel Golkowski, Thomas Künsting, 689-693. Jens Witsch

149 Rolf Sprengel Current Research The role of glutamate receptors in hippocam- Our major goal is to monitor pus mediated learning, emotional behaviors and to visualize molecular and mood disorders changes which underlie learning and memory Research Summary processes in the adult mouse brain. We use different NMDA receptor induced increase of AMPA currents at learning paradigms in the hippocampal CA1 cell synapses is one basic mechanism mouse and conditional underlying spatial learning and memory in the rodent. By ‘Knockout’ mouse models, genetic removal of NMDA and AMPA receptor subtypes which are impaired in in specific brain regions we dissect different forms of different forms of learning memories to identify underlying physiological processes. as basic tools in our studies. In the genetic background of these learning impaired mice we introduced by transgenic Curriculum Vitae technologies or by virus mediated gene transfers indicator Degrees: 1981 M.Sc. (Diploma), University of Heidelberg genes, which are then used to monitor the molecular 1983 Ph.D. University of Heidelberg changes before, during and after learning in brain slices 1983-1996: Postdoctoral fellow, Department of Microbiology, and in two photon live imaging. Primary neuronal cultures University of Heidelberg; Department of and organotypic slice cultures are used for in vitro studies. Microbiology and Immunology, University of California, San Francisco, USA; MPI for Biochemistry, Munich; Center for Molecular Biology, Memory deficiencies coupled to selective AMPA or University of Heidelberg NMDA receptor dependent plasticity (in defined princi- 1996-2001: Research associate, Department of Molecular pal cell layers of the hippocampus) Neurobiology, MPI for Medical Research, Heidel- berg NMDA receptor induced increase of AMPA currents at since 2001: Research Group leader, Department of Molecular Neurobiology, MPI for Medical Research, Heidel- hippocampal CA1 cell synapses is one basic mechanism berg underlying spatial learning and memory in the rodent. By genetic removal of NMDA and AMPA receptor subtypes in Contact specific brain regions we are able to dissect different forms Department of Molecular Neurobiology of memories and to identify underlying physiological Max Planck Institute for Medical Research processes. With global GluR-A knockout mice we made a Jahnstr. 29 provocative finding that spatial reference memory can be 69120 Heidelberg formed without spatial working memory. To understand Germany both memory forms at the physiological and molecular A +49-6221-486101 level in more detail we would like to approach the following E +49-6221-486119 major questions: K [email protected] o http://www.mpimf-heidelberg.mpg.de7forschungsgruppen/ • Can we dissect different forms of memory formation in sprengel/index.html fear conditioning? • Can we detect physiological parameters that are correlated with either spatial working or spatial reference memory?

150 Rolf Sprengel

• How can a spatial reference memory be formed without pathways that have been postulated for the pathogenesis working memory? or pathophysiology are investigated. • How is LTP connected to information storage?

Fig. 2: The gene for the NMDA receptor subunit NR1 is specifically Fig.1: Memory performance of mice is analyzed in (A) T-maze, (B) Y-maze removed in the gyrus dentatus of the hippocampus. (A) The expression of and in (C, D) conditioning chambers. Cre recombinase is restricted to the granular cells of the gyrus dentatus by the CamK2A/Grin2C promoter which controls the transcription factor (itTA) that is needed for Cre expression. In Cre expressing cells the gene The role of glutamate and glutamate receptors in mouse segments which are flanked by loxP sites (black triangles) are removed. models for emotional behaviors and mood disorders The loss of intact NR1 expression in the gyrus dentatus region can be monitored by in situ hybridization (B) or by anti-NR1 antibodies in The neurotransmitter glutamate is involved in the immunocytochemical stainings (C, lower panels). (C, upper panels) The pathophysiology and treatment of mood disorders. The gyrus dentatus specific Cre expression is visualized by anti-Cre antibodies goal of our studies is to dissect the role of glutamate in which show a strong labeling of the granular cell layer. Membrane depolarization results in a transient increase of pHluorin fluorescence mouse models for affective disorders. We subject different at single vesicle fusion sites (compare left panel with 2 right panels, s = genetically modified mice with impaired glutamate seconds). homeostasis and glutamate signaling to behavioral, neuroendocrinological, molecular and pharmacological Functional changes in neuronal networks in mice with studies. A variety of mutant mice with specific deficits in selective loss of AMPA or NMDA receptors glutamate uptake (EAAT-1 and EAAT-2 transporters) or glutamatergic neurotransmission, e.g. of ionotropic (AMPA- We have developed viral-mediated gene expression and NMDA-type) glutamate receptors (GluR-A, GluR-C, NR- systems for rapid and precise delivery of proteins, including 1, NR-2A) will be analyzed. By restricting the gene deletions genetically-encoded fluorescent calcium indicator to specific areas in the brain we will identify the relevant proteins (FCIPs), into neurons for recording activity at a brain regions. Glutamatergic drugs that shape emotional population scale. Our goal is to characterize by optical behavior in murine depression models will also be tested imaging spatiotemporal activity patterns of a group of in these mice. neurons activated by sensory experience. We will try to establish the essential role of NMDA- and AMPA receptors • Alterations in molecular/biochemical/cellular signaling in map plasticity.

151 Rolf Sprengel

Transcriptional and posttranscriptional modification of participate in trafficking, stability and synaptic localization AMPA receptor complexes of AMPA receptors will be quantified in specific regions of the brain. • Are changes in AMPA receptor signaling correlated with the development of addictive behavior?

Top publications • Jensen, V., et al. (2003). A juvenile form of postsynaptic hippocampal long-term potentiation in mice deficient for the AMPA receptor subunit GluR-A. J. Physiol. 553, 843-856. • Krestel, H. E., et al. (2004). A genetic switch for epilepsy in adult mice. J. Neurosci. 24, 10568-10578. • Schmitt, W. B., et al. (2005). Restoration of spatial working memory by genetic rescue of GluR-A-deficient mice. Nat. Neuros- ci. 8, 270-272. • Shimshek, D. R., et al. (2005). Enhanced odor discrimination and impaired olfactory memory by spatially controlled switch of AMPA receptors. PLoS Biol. 3, e354. • Sanchis-Segura, C., et al. (2006). Involvement of the AMPA re- Fig. 3: Negative EM stain of rAAV particles used for injection. Empty ceptor GluR-C subunit in alcohol-seeking behavior and relap- particles are donut-shaped. Scale 100 nm. se. J. Neurosci. 26, 1231-1238. We investigate AMPA receptor complexes from brain tissue • Shimshek, D., et al. (2006). Forebrain specific GluR-B deletion of rats addicted to alcohol. Particularly, the expression impairs spatial memory but not hippocampal field LTP. J. Neu- rosci. 26 (33), 8428-8440. and postranscriptional modification of genes which • Humeau, Y., et al. (2007). A pathway-specific function for different AMPA receptor subunits in amygdala long-term potentiation and fear conditioning. J. Neurosci. 27, 10947-10956. • Zhu, P., et al. (2007). Silencing and Un-silencing of Tetracycline- Controlled Genes in Neurons. PLoS ONE 2, e533. • Engblom, D., et al. (2008). Glutamate Receptors on Dopamine Neurons Control the Persistence of Cocaine Seeking. Neuron 59, 497- 508. • Wallace, D., et al. (2008). Single-spike detection in vitro and in vivo with a genetic Ca2+ sensor. Nat Meth. 5, 797- 804.

Structure of the Group Group Leader: Rolf Sprengel Postdoctoral fellows: Simone Giese, Liliana Layer PhD-Students: Thorsten Bus, Godwin Dogbevia, Wannan Tang, Yiwei Cheng, Dario Arcoz-Diaz Fig. 4: Unilateral cortex and hippocampus injection of rAAV encoded Technicians: Hans Gaugler, Annette Herold, Venus fluorescent protein ten days after injection. Scale 0.4 mm. Simone Hundemer, Martina Lang

152 Current Research Kerry L. Tucker An essential role for primary Early events in neurogenesis and nerve deve- cilia in the development of lopment in the central and peripheral nervous the forebrain systems Primary cilia have recently been implicated as crucial Research Summary sites of signal transduction Our laboratory is concerned with several aspects of and protein processing that neurogenesis and nerve outgrowth in the developing are important for a wide murine nervous system. In the cortex, we are working range of developmental with several mutants involved in the trafficking of functions (Fig. 1). We developmentally-relevant signalling molecules within have found an essential role of primary cilia in forebrain primary cilia. We are also investigating the role that development. The recessive mutation cobblestone (cbs) histone deacetylases play in the development of the forebrain. In the periphery, we employ an organotypic slice culture system to manipulate and image spinal nerve innervation into the forelimb.

Curriculum Vitae Degrees: 1990 B.A. at Harvard College, Cambridge, USA 1997 Ph.D. Massachusetts Institute of Technology, Cambridge, USA 1993-1997: Ph.D. research, Whitehead Institute for Biomedical Research, Cambridge, USA 1997-2003: Postdoctoral fellow, MPI for Neurobiology, Martinsried, Germany and Friedrich Miescher Institute, Basel, Switzerland 2003: Sabbatical, Biogen Idec, Inc, Cambridge, USA Since 2003: Junior Group Leader, IZN, University of Heidelberg, Germany

Contact Institute of Anatomy University of Heidelberg Im Neuenheimer Feld 307 69120 Heidelberg Germany A +49-6221-548687 E +49-6221-544952 Fig. 1: Primary cilia project into the dorsolateral telencephalic ventricles K [email protected] (V) of 12.5 d.p.c. mouse embryos. (A,B) Transmission electron microscopy o http://www.izn.uni-heidelberg.de/ showing a cilium cut longitudinally (A) and in cross section (B), the latter revealing the characteristic “9 + 0” ciliary morphology. (C) Scanning electron microscopy of cilia (indicated by arrows) lining the ventricle (V) of embryonic brain. Scale bars: A,B: 200 nm, C: 1000 nm.

153 Kerry Tucker

and they have recently been implicated in a diversity of postnatal functions and dysfunctions in the brain including ocular dominance plasticity, the consolidation of memory in the hippocampus, epileptic seizures, and depression. We have investigated their role in the development of neurons in embryonic mouse brain. As a wide variety of HDACs are expressed in differentiating neural progenitor Fig. 2: Coronal slices of 14.5 d.p.c brains from normal (left) and cbs -/- cells, we have taken a pharmacological approach to mutant (right) embryos, stained with hematoxylin. Arrows indicate inhibit multiple family members. Inhibition of class I and evaginated cortex in the mutant. Scale bar = 1 mm. II HDACs in developing mouse embryos with trichostatin A resulted in a dramatic reduction in neurogenesis in the was identified in an ENU-based mouse mutagenesis screen striatum and a modest increase in neurogenesis in the and is distinguished by cortical “heterotopias”, appearing cortex. A reduction in neurogenesis in striatum-derived at 10.5 d.p.c. as protrusions from the pial surface of the neural precursors was accompanied by an increase in dorsal forebrain. Homozygous mutants showed defects the production of immature astrocytes. We have shown in the formation of the dorsomedial telencephalon, the that HDACs control neurogenesis by inhibition of the evagination of folds at the ventricular zone of the cortex bone morphogenetic protein BMP2/4 signaling pathway. and thereby to the formation of the heterotopias (Fig. 2). HDACs function at the transcriptional level by inhibiting Later embryonic stages show a failure to form superficial and promoting, respectively, the expression of Bmp2 cortical layers such as the subplate, the cortical plate and and Smad7, an intracellular inhibitor of BMP signaling. the marginal zone. Inhibition of the BMP2/4 signaling pathway restores normal In situ analysis indicated the presence of the choroid levels of neurogenesis and astrogliogenesis to both striatal plexus, cortical hem and hippocampal anlage, but the and cortical cultures in which HDACs are inhibited. Our morphology of these structures was severely disturbed. In results demonstrate a transcriptionally-based regulation contrast, development of the ventral forebrain appeared of BMP2/4 signaling by HDACs both in vivo and in vitro largely normal. Standard genetic mapping approaches that is critical for striatal neurogenesis and that modulates and classical complementation analysis have identified cbs as a mutation in the Ift88 gene. The Ift88 protein plays an important role in the construction and maintenance of both primary and motile cilia, acting in a complex that is responsible for the intraflagellar transport of protein cargos. Northern blot, real time RT-PCR, and Western blot analysis showed a 70-80% decrease in levels of the Ift88 mRNA and protein, respectively, indicating a hypomorphic allele. Altered processing of the transcription factor Gli3 has been observed in the brains of cbs mutants, while other developmental pathways causing this pleiotropic phenotype are currently under investigation.

Histone deacetylases control neurogenesis in embryonic brain Histone-modifying enzymes are essential for a wide variety of cellular processes dependent upon gene regulation. Fig. 3: Transverse slice from tauGFP embryo. The dorsal root ganglion (D) and the motor neurons (M) of the spinal cord together contribute to Histone deacetylases (HDACs) lead to the compaction of the spinal nerves (lower half of figure) growing into the forelimb. Scale chromatin and subsequent silencing of gene transcription, bar = 500 µm.

154 Kerry Tucker cortical neurogenesis. This suggests that HDACs may Top publications regulate the developmental switch from neurogenesis • Benninger, F., et al. (2003). Functional integration of embryo- to astrogliogenesis that occurs in late gestation. We are nic stem cell-derived neurons in hippocampal slice cultures. J. pursuing this question through identification of the Neurosci. 23, 7075-7083. responsible HDAC genes, their interaction partners, and • Andorfer, C., et al. (2003). Hyperphosphorylation and aggrega- their molecular targets. tion of tau in mice expressing normal human tau isoforms. J. Neurochem. 86, 582-590. Imaging of peripheral nerve outgrowth in slice culture • Plachta, N., et al. (2004). Developmental potential of defined neuronal progenitors derived from mouse embryonic stem Newborn neurons elaborate an axon that undertakes a cells. Dev. 131, 5449-5456. complicated journey to find its ultimate target in the brain • Bibel, M., et al. (2004). Differentiation of mouse embryonic or periphery. Although major progress in the study of this stem cells into a defined neuronal lineage. Nat. Neurosci. 7, process has been made by examination of dissociated 1003-1009. neurons in vitro, one would like to observe and manipulate • Korets-Smith, E., et al. (2004). Cre recombinase specificity defi- axonal outgrowth and pathfinding as it occurs in situ, as ned by the tau locus. Genesis 40, 131-138. fasciculated nerves growing within the tissue itself. We • Wernig, M., et al. (2004). Functional integration of embryonic have developed a simple technique to do this, through stem cell-derived neurons in vivo. J. Neurosci. 24, 5258-5268. cultivation of embryonic mouse slices from the tauGFP • Gao, L., et al. (2005). Transcriptional regulation of the mouse mouse line that expresses EGFP specifically in newborn microtubule-associated protein tau. Biochim. Biophys. Acta neurons. This system allows for imaging of outgrowth of 1681, 175-81. peripheral nerves into structures such as the developing • Brachmann, I., et al. (2007). A simple slice culture system for limb (Fig. 3). the imaging of nerve development in embryonic mouse. Dev. With this technique we can reproduce normal innervation Dyn. 236, 3514-3523. patterns by spinal nerves derived from spinal cord motor • Willaredt, M.A., et al. (2008). A crucial role for primary cilia in cortical morphogenesis. J. Neurosci. 28, 12887-12900. neurons and sensory neurons of the dorsal root ganglia. • Shakèd, M., et al. (2008). Histone deacetylases control neuro- The slices can be manipulated pharmacologically as well genesis in embryonic brain by inhibition of BMP2/4 signaling. as genetically, by crossing the EGFP-expressing line with PLoS ONE 3(7), e2668. doi:10.1371/journal.pone.0002668. lines containing targeted mutations in genes of interest. • Wolff, S.B.E., et al. (2008). Die Rolle der Histon-Acetylierung für We are exploiting the latter possibility with a mutant in Lernen und Gedächnis. Neuroforum 14, 274-279. the gene encoding Semaphorin3A, a key regulator of early spinal nerve outgrowth.

Transgenic mouse lines expressing modulators of Rho GTPase function We have generated a series of mouse lines in which all postmitotic neurons express modulators of Rho GTPase function. The first line we have derived expresses a Structure of the Group dominant negative inhibitor of RhoA in which amino Group Leader: Kerry Tucker acid 19 has been substituted to asparagine (N19-RhoA). Visiting Scientists: Dean O. Smith, Sidney Cambridge Through an unknown mechanism, this results in an PhD-Students: Isabel Brachmann, Christian Gojak, overrepresentation of neurons in all layers of the cerebral Kathrin Weissmüller, Marc Willaredt cortex, when analyzed at maturity. As we have not found Undergraduates: Kristin Bobsin, Valentin Evsyukov, defects in layering identity so far, we believe the cause to lie Silke Herzer, Veronika Kremer, in the regulation of developmentally-occurring postnatal Hannah Meyer, Dmitry Rusanov, apoptosis, and this is currently being examined. Xiao-Rui Sun, Xiao Shen, Markus Stahlberg

155 Klaus Unsicker Current Research TGF-ß, FGF and neurotrophins in neural deve- Generation of cell diversity lopment and functions in the sympathoadrenal cell lineage of the neural Research Summary crest We are interested in the functions of three major families The neural crest (NC) gives of growth factors in the developing and adult nervous rise to different types of system. We focus on the development of neural crest neurons, glial, endocrine, derivatives, functions of limbic areas and mesostriatal/ and mesenchymal cells and, mesolimbic systems in health and disease. An important hence, is an excellent model topic in the research of the laboratory is the molecular for exploring mechanisms underlying the generation of cell diversity. We focus understanding of neuronal survival and death. on neuroendocrine (chromaffin) derivatives of the NC Methodologies include mouse mutants, cell and tissue (see Fig. 1). We have shown that, contrary to a classic culture, biochemistry, molecular biology, histology, and hypothesis, glucocorticoid hormones and adrenal cortical electrophysiology. cells are not required for most aspects of chromaffin cell differentiation [1, 2]. Currently, we study the role of Curriculum Vitae BMP-4 and its receptors in the induction of chromaffin Degrees: 1968 MD in Histology, University of Kiel cell fate and differentiation. BMP-4 is an essential factor 1969-1975: Postdoctoral fellow, Kiel, Lund, Sweden, for inducing a catecholaminergic phenotype in NC cells. Melbourne, Australia, UCL, London, UK It is secreted by cells in the wall of the dorsal aorta and 1978-1992: Full Professor and Head of Department, surrounding mesenchyme, rapidly downregulated in Anatomy and Cell Biology, University of Marburg sympathetic ganglionic anlagen, but persistent in adrenal 1983-1984: Visiting Professor, Department of Biology, cortical cells [7]. This raised the question whether BMP-4 University of California, San Diego, USA might be important in the induction and differentiation 1989-1990: Visiting Professor, Laboratory of of the chromaffin phenotype. In a series of loss- and gain- Chemoprevention, NCI, NIH, Bethesda, USA 2006: Visiting Professor, Institute of Biotechnology, University of Helsinki, Finland since 1992: Full Professor and Director, Department of Anatomy and Cell Biology, Unversity of Heidelberg since 2000: Director, IZN, University of Heidelberg

Contact Institute of Anatomy and Cell Biology Department of Molecular Embryology University of Freiburg Alberstr.17 79104 Freiburg Germany A +49-761-2035193 E +49-761-2035091 K [email protected] Fig. 1: In situ hybridisation showing TH mRNA (red) in adrenal chromaffin o http://portal.uni-freiburg.de/anatomie2 cells and BMP4 mRNA (blue) in adrenal cortical cells of an E9 chick embryo.

156 Klaus Unsicker of-function experiments in chick embryos we found that 24h) suggesting that IGF-1 can positively and negatively BMP-4 did not switch neuronal to chromaffin cell fate, regulate the ERK pathway [4]. but significantly promoted further differentiation of chromaffin cells. By GFP electroporation into neural tube Functions of a novel TGF-ß in neuronal development and NC cells we try to analyze the temporal sequence of and maintenance delamination of distinct progenitor subpopulations. GDF-15 is a novel distant member of the TGF-ßs, originally Preliminary results suggest that the vast majority of single identified as a potent trophic factor for midbrain NC cells can give rise to both, sympathetic neurons and dopaminergic neurons. We have generated a GDF-15 neuroendocrine chromaffin cells. knockout mouse, which exhibits motoneuron and neural stem cell phenotypes. There is a progressive postnatal Development of a CNS-peripheral nervous system cir- loss of hindbrain and spinal cord motoneurons from 3 cuitry to 6 months of age resulting in an approx. 20% deficit. In Neurons located in the lateral spinal cord and in the addition, mutant mice have revealed hypermyelination of brainstem link the central and peripheral portions of the axons and increased expression of myelin proteins. autonomic nervous system. These neurons innervate peripheral autonomic ganglia and chromaffin cells. Their trophic regulation is not well understood. We have recently discovered that cardiotrophin-1 and the LIFRß are crucial for the postnatal maintenance of this neuron population [3]. By comparing neurons of the pre- and paravertebral sympathetic ganglia we found marked differences, as e.g. in NGF dependence. It is possible that prevertebral in contrast to paravertebral ganglia receive a second neurotrophic signal from their innervation targets (enteric nervous system and gut). If so, neurons in the prevertebral ganglia should, in addition to trkA, express a further neurotrophic factor receptor. To test this, we currently started an in situ Fig. 2: Alpha-synuclein accumulating neurons in the SNpc of an aged hybridization analysis using different probes for trophic trkB/C(+/-)/(+/-) mouse (cell nuclei are stained in blue by DAPI; alpha-synu- factor receptors (e.g. neurotrophin receptors and GFR-α clein immunoreactivity is shown in red). receptors). This would be consistent with Schwann cells as a source Neuron survival and the roles of ERK and a role of GDF-15 in regulating myelination. However, mechanisms underlying motoneuron death still remain to A classic perception of the molecular bases of neuron be explored. Neural stem cells (NSC) express high levels death implies ERK as an important regulator of neuron of GDF-15 mRNA, and GDF-15 KO mice have less EGFR survival. We have now found that ERK has also a key role expressing NSC that wt mice. Details of GDF-15 functions in promoting neuronal death of cerebellar granule cells in NSC proliferation and differentiation are currently under induced by K+ withdrawal [4, 5]. Specifically, neuronal death investigation. mediated by plasma membrane damage is accompanied by a late and sustained activation of ERK. Inhibition of TrkB and trkC neurotrophin receptors: spine morpholo- ERK activation results in a dramatic reduction of neuron gy and dopamine neurons death. Classic growth factors, as e.g. IGF-1, induce an early and transient ERK activation (15min to 2h), and also Current analyses of adult and aged heterozygous trkB and abrogate the appearance of late and sustained ERK (6 to trkC mice as well as of young adult conditional CamKII-

157 Klaus Unsicker

trkB knockout mice have revealed the importance of these inhibits extracellular signal-regulated kinase to promote neu- receptors for the maintenance of dendritic spine numbers ronal survival via the Phosphatidylinositol-3 kinase/Protein and dendritic spine morphology in the hippocampal field kinas-A/c-Raf pathway. J. Neurosci. 25, 2838-2852. CA1 [6]. • Gut, P., K. et al. (2005). Lack of an adrenal cortex in /Sf1/ mutant These analyses of adult and aged heterozygous trkB and mice is compatible with the generation and differentiation of trkC mice also showed that these receptors are crucially chromaffin cells. Dev. 132, 4611-4619. important for the maintenance of the dopaminergic • Bohlen und Halbach, O.v., et al. (2005). Haploinsufficiency for trkB and trkC receptors induces cell los and accumulation of a- nigrostriatal system in aged mice. The aged heterozygous synuclein in the substantia nigra. FASEB J. 19, 1740-1742. (see knockout mice showed a reduction in the number of online article of July 21, 2005: 10.1096/fj.o5-3845fje.) dopaminergic neurons in the substantia nigra (SN). This • Bohlen und Halbach, O.v., et al. (2006). Regional- and age- was accompanied by a massive accumulation of alpha- dependent reduction in trkB receptor expression in the hip- synuclein in the remaining neurons of the SN (Fig. 2), pocampus is associated with altered spine morphologies. Biol. underscoring the significance of trk mediated signaling Psychiat. 59, 793-800. for preventing degeneration and death of this neuron • Oberle, S., et al. (2006). Loss of leukemia inhibitory factor re- population. ceptor ß or cardiotrophin-1 causes similar deficits in pregan- [1] Finotto, S., et al. (1999) Development 126, 2935 – 2944 glionic sympathetic neurons and adrenal medulla. J. Neurosci. [2] Gut, P., et al. (2005) Development 132, 4611 – 4619 26, 1823-1832. [3] Oberle, S., et al (2006) J. Neurosci. 26, 1823 - 1832 • Peterziel, H., et al. (2007). Specificity in the crosstalk of TGFß/ [4] Subramaniam, S., et al. (2004) J. Cell Biol. 165, 357 – 369 GDNF family members is determined by distinct GFR alpha re- [5] Subramaniam, S., et al. (2005) J. Neurosci. 25, 2838 – 2852 ceptors. J. Neurochem. 103, 2491-2504. [6] von Bohlen und Halbach, O., et al. (2006) Biol. Psychiatry. 59, • Schober, A., et al. (2007). GDNF applied to the MPTP-lesioned 793-800 nigrostriatal system requires TGF-ß for its neuroprotective ac- [7] Huber, K., et al. (2008) Neural Development 3: 28 tion. Neurobiol. Dis. 25, 378-391. • Huber, K., et al. (2008). Persistent expression of BMP-4 in embryonic chick adrenal cortical cells and its role in chromaffin cell development. Neural Dev. doi:10.1186/1749-8104-3-28. • von Bohlen und Halbach, O., et al. (2008). TrkB but not TrkC receptors are necessary for postnatal maintenance of hippocampal spines. Neurobiol. Aging 29, 1247-1255. Top publications

• Farkas, L.M., et al. (2003). Transforming growth factor-ßs are Structure of the Group essential for the development of midbrain dopaminergic neurons in vitro and in vivo. J. Neurosci. 23, 5178-5186. Group Leader: Klaus Unsicker • Reuss, B., et al. (2003). Functions of fibroblast growth factor Senior scientists: Oliver von Bohlen und Halbach, (FGF)-2 and FGF-5 in astroglial differentiation and blood-brain Katrin Huber, Andreas Schober barrier permeability: Evidence from mouse mutants. J. Neu- Postdoctoral fellows: Alexei Egorov, Jojanneke Huck, rosci. 23, 6404-6412. Heike Peterziel, Jens Strelau • Subramaniam S., et al. (2004). ERK activation promotes neuro- PhD-Students: Carmen Carillo-Garcia, Brandon Cline, nal degeneration predominantly through plasma membrane Krithi Irmady, Jarek Jarosik, damage and independently of caspase-3. J. Cell Biol. 165, 357- Lakshmi Kumaraswany, Tina Paech, 369. (See also Commentary by Cheung and Slack in Science Marie Schier, Sandra Werner STK2004, pe45.) MD Students: Sonja Krause, Sebastian Prochnow, • Medina, D.L., et al. (2004). TrkB regulates neocortex formation Sabrina Zechel through the Shc/PLC-mediated control of neuronal migration. Technicians: Gerald Bendner, Barbara Brühl, Jutta Fey, EMBO J. 29, 3803-3814. Richard Hertel, Ursula Hinz, • Subramaniam S., N. et al. (2005). Insulin-like growth factor 1 Petra Viforkas-Marzi

158 Current Research Wolfgang Wick Biology and experimental therapy of malig- Hematopoietic stem cells nant glioma as vehicles for a gliomas- directed therapy Research Summary This project aims at The putative origin of malignant gliomas from neuronal or elucidating the molecular other brain-derived stem cells challenges several aspects mechanisms of lesion- of current concepts in brain tumor biology and therapeutic tropism of hematopoietic approaches. Further key propensities of gliomas, such as stem cells (Tabatabai et al. migration and invasiveness as well as angiogenesis need 2005 & 2006). Independent further molecular clarification. Consequent experimental subprojects in this project are therapies involve vaccination approaches and delivery by hematopoietic and mesenchymal stem cells. • mechanism of endothelial transvasation of hematopoietic stem cells Curriculum Vitae • comparison of hematopoietic and mesenchymal stem Degrees: 1998 MD cells for the delivery of therapeutic molecules 2003 Habilitation • development of a cell-based antiangiogenic therapy for 1994-1997: Thesis work, Dep. of Neuropathology, malignant gliomas University of Bonn • optimization of an anti-transforming growth factor 1998-2003: Postdoctoral fellow and resident in Neurology, (TGF)-b therapy with stem cells Dep. of Neurology, University of Tübingen 2004-2006: Group Leader of the Neurooncology Group, Tübingen Functional characterization of invasion-related proteins 2003-2007: Head of the Research Group Hematopoietic Stem induced by chronic non-lethal hypoxia Cells and Glioma of the State of Baden- Malignant gliomas are hypoxic tumors and this contributes Württemberg 2007: Full Professor, University Hospital of Heidelberg, to the resistance to therapy and putatively also to the Neurooncology invasive phenotype. In a proteome study several proteins have been identified to be differentially expressed under Contact hypoxic treatment compared to normoxic conditions and could be confirmed by orthogonal methods. Of Department of Neurooncology University Clinic of Heidelberg these candidates N-myc downstream regulated gene Im Neuenheimer Feld 400 1 protein (NDRG1) and neuroenolase (ENO2) will be 69120 Heidelberg analysed. Silencing of the corresponding candidate genes Germany and reexposing these cells to hypoxia in the invasion/ A +49-6221-567075 apoptosis and angiogenesis paradigms will underscore the E +49-6221-567554 functional relevance of the candidates for the induction of K [email protected] an invasive phenotype or gliomas angiogenesis. Applying o http://www.klinikum.uni-heidelberg.de/neuroonkologie proteomics techniques, additionally involved proteins are looked for in cells harboring the silenced candidate gene to find downstream molecules that potentially serve as therapeutic targets for an anti-angiogenic therapy.

159 Wolfgang Wick

Novel functions of BCL-2 family proteins in glioblasto- • Development of a protein kinase C inhibitor for the mas - invasiveness and autophagy therapy of malignant gliomas (Tabatabai et al. 2007) Since this subproject focuses on invasion analyses, and experimental autoimmune encephalomyelitis. apoptotic and non-apoptotic cell death mechanisms, • Evaluation of anti-glioma vaccination. and orthotopic glioma animal models. In glioblastomas, • Description and functional characterization of gliomas- the BCL-2 and BCL-xL proteins contribute to an overall initiating cells. resistance to apoptosis which correlates with the clinically relevant chemo- and radioresistance. Our recent data demonstrate that BCL-xL specifically promotes the invasiveness of glioma cells in vitro and in vivo (Weiler et al. 2006). This novel BCL-xL function is mediated by altered gene expression and does not depend on the direct anti- apoptotic effect of BCL-xL through sequestering pro- apoptotic BCL-2 family members. Moreover, BCL-2 and BCL-xL were recently described to be major regulators of autophagic cell death (type II cell death). There is increasing evidence that autophagy plays a crucial role in the tumorigenesis and therapy resistance of glioblastoma.

Fig. 2: Free floating human glioblastoma tumor spheres of the tumors 323 and 325 cultured in NSC-M after excision and homogenisation and respective adherent non-sphere-forming cells.

Fig. 1: Adult hematopoietic stem cell exhibit a TGF-β/CXCL-12-dependent tropism for gliomas in vitro and in vivo. Images 1 - 3 depict an experimental LNT-229 intracranial glioma (nuclei DAPI stained (1-2) or H&E staining (3)) invaded by hematopoietic stem cells (PKH26). Image 4 shows that there are no stem cells in the non-tumor hemisphere.

160 Wolfgang Wick

Top publications • Friese, M.A., et al. (2004). RNA interference targeting TGF-β1,2 enhances NKG2D-mediated anti-glioma immune response, inhibits glioma cell migration and invasiveness and abrogates tumorigenicity in vivo. Cancer Res. 64, 7596-7603. • Tabatabai, G., et al. (2005). Lessons from the bone marrow: how malignant glioma cells attract adult hematopoietic stem cells. Brain 128, 2200-2211. • Weiler, M., et al. (2006). Bcl-xL, apoptosis and motility: move to survive or survive to move? Cell Death Differ. 13, 1156-1169. • Tabatabai, G., et al. (2006). Irradiation and hypoxia promote homing of hematopoietic progenitor cells towards gliomas by TGF-β-dependent HIF-1α-mediated induction of CXCL12. Brain 129, 2426-2435. • Tabatabai, G., et al. (2007). Synergistic antiglioma activity of radiotherapy and enzastaurin. Ann. Neurol. 61, 153-161. • Roth, P., et al. (2007). Malignant glioma cells counteract an- titumor immune responses through expression of lectin-like transcript-1.Cancer Res. 67, 3540-3544. • Wick, A., F et al. (2007). Efficacy and tolerability of Temozo- lomide in an one week on/one week off regimen in patients with recurrent gliomas. J. Clin. Oncol. 25, 3357-3361. • Murat, A., et al. (2008). Stem cell-related „self-renewal signature“ and high EGFR expression are associated with resistance to concomitant chemoradiotherapy in glioblastoma. J. Clin. Oncol. 26, 3015-3024. • Tabatabai, G., et al. (2008). Glioma cell-mediated upregulation of CD62E on endothelial cells contributes to glioma tropism of adult hematopoietic progenitor cells. Brain, Epub. • Wick, W., et al. (2008). A novel tool to analyse MRI recurrence patterns in glioblastoma. Neuro-oncol, Epub.

Structure of the Group Group Leader: Wolfgang Wick Senior scientist: Michael Platten PhD Students: Ulrike Litzenburger, Christiane Optiz, Phillip Pfenning, Markus Weiler, Lorna Whyte Undergraduates: Sebastian Boch, Anke Funke-Kaiser, Sebastian Luger Technicians: Petra Rübmann

161 Otmar D. Wiestler Research interests Molecular neuropathology of human brain Surgical and Molecular tumors / surgical neuropathology of CNS Neuropathology of Human tumors Brain Tumors Based on a large number Research Summary of referrals to The German • Molecular Neuropathology of Human Brain Tumors Brain Tumor Reference • Surgical Neuropathology of CNS Tumors Center at the University • Reconstructive Neurobiology of Bonn Medical Center, • Molecular Neuropathology of Focal Human Epilepsies our group has carried out extensive studies on the Curriculum Vitae morphological and molecular characterization of human CNS neoplasms. Critical information has also been derived Degrees 1984 MD, University of Freiburg from therapeutic trials for which the center served as 1984-1987: Postdoctoral fellow at the Department of Pathology, University of California, San Diego, reference neuropathology laboratory. USA The 2007 WHO Classification of Brain Tumors, for which 1987-1989: Residency training in Neuropathology, Institute of the consensus conference was held in Heidelberg in 2006, Neuropathology, Department of Pathology, serves as the international standard for the diagnosis and University of Zürich, Switzerland characterization of nervous system cancers. 1989-1992: Senior resident in Neuropathology, Institute of Neuropathology, Department of Pathology, University of Zürich, Switzerland 1992-2003: Professor of Neuropathology and Head of the Department of Neuropathology, Medical Center, University of Bonn 1994-2003: Head of the German Brain Tumor Reference Center since 2004: Chairman and Scientific Member of the Management Board, German Cancer Research Center (DKFZ), Heidelberg

Contact German Cancer Research Center Im Neuenheimer Feld 280 69120 Heidelberg Germany A +49-6221-422850 E +49-6221-422840 K [email protected] o http://www.dkfz.de

Fig. 1: Ganglioglioma with binucleated neuron and strong immunoreactivity for synaptophysin.

162 Otmar Wiestler

Molecular Neuropathology of Human Gliomas and Top publications Medulloblastomas • Brüstle, O., et al. (1999). Embryonic stem cell-derived glial pre- In cooperation with Andreas von Deimling (Heidelberg) cursors: a source of myelinating transplants. Science 285, 754- and Torsten Pietsch (Bonn), human gliomas and 756. medulloblastomas have been subjected to a systematic • von Deimling, A., et al. (2000). Comprehensive allelotype and genetic analysis of 466 human nervosu system tumors. J. Neu- molecular and cellular analysis. The studies yielded ropathol. Exp. Neurol. 59, 544-558. important novel insights into molecular pathways in the • Mueller, W., et al. (2002). Genetic signature of oligoastrocyto- formation of these tumors some of which have been mas correlates with tumor location and denotes distinct mo- introduced in the diagnostic repertoire. lecular subsets Am. J. Pathol. 161, 313-319. In the case of childhood medulloblastomas, a combination • Scheffler, B., et al. (2003). Functional network Integration of of molecular and cell biological experiments made it Embryonic stem cell-derived astrocytes in hippocampal slice possible to unravel the origin of these embryonal tumors cultures Development 130, 5533-5541. from distinct cerebellar progenitors and to associate these • Waha, A., et al. (2003). Epigenetic silencing of the HIC-1 tumor subtypes with distinct molecular alterations. suppressor gene in human medulloblastomas. J. Neuropathol. Exp. Neurol. 62, 1192-1201. • Thal, D.R., et al. (2003). Vascular pathology in Alzheimer’s di- Molecular Neuropathology of Epilepsy-associated sease: correlation of cerebral amyloid angiopathy and arte- Neural Tumors riosclerosis/lipohyalinosis with cognitive decline. J. Neuropa- Patients with chronic, focal epilepsies often display thol. Exp. Neurol. 62, 1287-1301 • Felsberg, J., et al. (2004). Oligodendroglial tumors: Refinement low-grade neoplasms with distinct morphological of candidate regions on chromosome arm 1p and correlation and molecular properties. Characteristics include high of 1p/19q status with survival. Brain Pathol. 14, 121-130. differentiation, presence of both glial and neuronal • Hartmann, W., et al. (2006). Phosphatidylinositol 3’-kinase / components, a potential origin from malformative lesions Akt signaling is activated in medulloblastoma cell prolifera- and a pattern of molecular changes distinct from other tion and is associated with reduced expression of PTEN. Clin. CNS neoplasms. Cancer Res. 12, 3019-3027. • Weber, R.G., et al. (2007). Frequent loss of chromosome 9, homozygous CDKN2A/p14ARF/CDKN2B deletion and low TSC1 mRNA expression in pleomorphic xanthoastrocytomas. Onco- gene 26, 1088-1097. • Trumpp, A., Wiestler, O.D. (2008). Mechanisms of Disease: cancer stem cells – targeting the evil twin. Nat. Clin. Pract. Oncol. 5 (6).

163 Joachim Wittbrodt Current Research Vertebrate retina development and While our focus in the first differentiation period at EMBL (1999- 2002) was mainly on early Research Summary pattering, the second period In vertebrates, patterning of the anterior neural plate culminates (2003-2007) is characterized in the defined expression of the evolutionarily conserved by key findings in optic transcription factors Six3 and Pax6. In the region where Six3 vesicle (Rembold et al., 2006) and Pax6 expression overlap, retinal fate is specified. Under and optic cup (see below) the influence of midline signalling the retinal anlage is split morphogenesis. into two retinal primordia. Six3 is also required in this process. The two retinal primordia require the homeobox containing transcription factor Rx3 for their evagination to form the optic Our gain of function experiments had indicated that the vesicle.Those are further subdivided to give rise to neural retina, homeodomain containing transcription factor Six3 is retinal pigmented epithelium and optic stalk respectively. In sufficient for ectopic eye formation (Loosli et al., 1999). anamniotes, the ciliary margin of the neural retina contains a In a loss of function analysis we showed that its activity stem cell population that gives rise to all retinal cell types and is also necessary for the formation of the entire forebrain facilitates live long growth of the eye. and eyes (Carl et al., 2002). Interestingly Six3 also plays Curriculum Vitae subsequent roles related to the proximo-distal patterning of the eye field. Our analysis indicates that, depending Degrees: 1987 M.Sc. on the interactor, Six3 determines different fates along 1990 Ph.D. the proximo-distal axis (Tessmar et al., 2002) and, by 1999 Habilitation (Cell Biology and Developmental Biology ) direct interaction with the replication initiation complex, Technical University of Braunschweig positively acts on proliferation (Del Bene et al., 2004). This 1987-1990: Thesis work, MPI for Biochemistry, Martinsried interaction interestingly shows the direct interference of a 1991-1994: Postdoctoral fellow, Basel Biocenter, Switzerland transcription factor with the cell cycle and conversely allows 1995-1998: Group Leader, MPI for Biophysical Chemistry, the cell cycle to modulate the transcriptional potential of a Göttingen key player in eye development. In the absence of a battery 1999-2007: Group Leader, EMBL Heidelberg of Six3 transcriptional targets the experimental basis for Since 2007: Director at the Institute for Genetics and this far-reaching possibility was however thin. We have Toxicology, Center of Research, Karlsruhe further pursued this issue and have identified Six3 target Since 2007: Full Professor, Institute for Zoology, University of Heidelberg genes by ChIP from FAC sorted Six3-positive cells out of the embryonic context. In collaboration with colleagues Contact form the Genome Institute of Singapore (Yijun Ruan, Chia- Department of Molecular Developmental Biology/Animal Lin Wei, GIS) we analyzed Six3 targets in a genome wide Physiologie survey and (among many others) could show the direct Institute for Zoology, Unversity of Heidelberg binding of Six3 to its own upstream regions. Im Neuenheimer Feld 230 69120 Heidelberg, Germany Eye Morphogenesis A +49-6221-545653 E +49-6221- Optic vesicle evagination, the subsequent step in eye K [email protected] development, is impaired in the eyeless (el, medaka) and o http://www.zoo.uni-heidelberg.de chokh (chk, zebrafish) mutants (Loosli et al., 2003; Winkler

164 Joachim Wittbrodt et al., 2000). The gene affected is another homeodomain individual cells in the context of eye formation (Rembold containing transcription factor, Rx3 (Loosli et al., 2003; et al., 2006). Loosli et al., 2001). Our analysis facilitated the generation of transgenic lines (in medaka and zebrafish) that express Eye Evolution GFP specifically in the eye field of wild type and mutant The work on Rx genes had led us to also investigate their embryos. These lines allowed following optic vesicle role in basal Prototomia. We found that in the larva of formation in vivo by 4D confocal analysis (Huisken et al., the basic Lochtrophozoan Platynereis, Rx is expressed in 2004; Rembold et al., 2006), one of our milestones in the ciliary photoreceptors, as in the eyes of Deuterostomia. past four years. In these studies we unraveled that organ These photoreceptors that are not part of the eye, are morphogenesis crucially depends of the migratory activity found in a complex network within the conserved axonal of cells that, in contrast to the text-book view, migrate as scaffold. This and other data led us to propose that the body plan of Urbilateria was quite elaborated (Arendt et al., 2001). It contained already a visual complex at the anterior end of a conserved axonal scaffold that served as the evolutionary substratum from which the eyes of Proto- and Deuterostomia emerged (Arendt et al., 2004).

Medaka Genomics To further develop medaka into a competitive model system in an efficient and economical way, it was of key importance to strengthen our contacts to the Japanese medaka community. We have been successfully collaborating with the Kondoh differentiation project and completed the first description of mutants from a joint mutagenesis screen published in the “Medaka issue” of Mechanisms of Development (June/July 2004). This

Fig. 1: Overview of the eye of a double transgenic medaka fish at 4 days post fertilization. Green cells are expressing a membrane localized YFP under the control of the ath5 promoter, labeling the retinal ganglion cell Fig. 2: Time-lapse montage of lyn-tandem tomato expressing cells in the layer. Ciliary marginal zone, Müller glia and photoreceptors are labelled retina under the control of the vsx3 promoter. Columns of precursor- in red by H2B-RFP under the control of the rx2-promoter. The image is a cells develop from a vertical to a horizontal network forming the inner maximum projection of a confocal stack recorded with the Leica SP5 and plexiforme layer in the retina. Cells were imaged live and in vivo using a a 20x glycerol objective. Leica SP5 and a 20x oil immersion objective.

165 Joachim Wittbrodt and other activities catalyzed the initiation of a genome Top publications project that covers all aspects of medaka genomics. The • Arendt, D., et al. (2004). Ciliary photoreceptors with vertebra- medaka genome has now been released and was been te-type opsins in an invertebrate brain. Science 306, 869-871. published in Nature. With the expected publication of the • Huisken, J., et al. (2004). Optical sectioning deep inside live zebrafish genome in the near future, almost 150 million embryos by selective plane illumination microscopy. Science years of evolution will be experimentally amenable in 305, 1007-1009. these fully developed model systems, putting fish into a • Del Bene, F., et al. (2004). Direct interaction of geminin and unique position among the vertebrates. Six3 in eye development. Nature 427, 745-749. • Rembold, M. et al. (2006). Individual cell migration serves as We have started to establish ourselves in the field of the driving force for optic vesicle evagination. Science 313, 1130-1134. bioinformatics to complement and direct bench work. At • Martinez-Morales, J.-R. et al. (2005). Differentiation of the Ver- the moment we focus on mining the genomic information tebrate Retina Is Coordinated by an FGF Signaling Center. Dev. available in the context of gene and genome evolution Cell 8, 565-574. (Martinez-Morales et al., 2007) and transcriptional networks • DelBene, F., et al. (2007). In vivo Validation of a Computatio- related to eye development and evolution (Ettwiller et al., nally Predicted Conserved Ath5 Target Gene Set. PLOS Genet. 2005; Henrich et al., 2003; Henrich et al., 2004; Henrich et 3, 1661-1671. al., 2005) in order to establish a virtuous cycle between • Ettwiller, L., et al. (2007). TRAWLER: de novo Regulatory Motif bench and computer to direct future experimental work in Discovery Pipeline for Chromatin Immunoprecipitation. Nat. a more system wide context. Meth., 563-565. • Grabher, C., et al. (2007). Birth and life of tissue macrophages and their migration in embryogenesis and inflammation in medaka. .J. Leukocyte Biol. 81, 263-271. • Keller, P. J., et al. (2008). Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy. Sci- ence 322, 1065-1069. • Ramialison, M., et al. (2008). Rapid identification of PAX2/5/8 direct downstream targets in the otic vesicle by combinatorial use of bioinformatics tools. Genome Biol 9, R145.

Structure of the Group Group Leader: Joachim Wittbrodt Postdoctoral fellows: Lazaro Centanin, Petra Haas, Daigo Inoue, Mirana Ramialison, Stephanie Schneider Fig. 3: Reconstruction of the zebrafish digital embryo. The right half of the embryo is shown as the microscopy data, where nuclei are labeled using PhD students: Annette Schmidt, Philipp Keller (joint with H2B-GFP (yellow/red LUT). The left half shows the segmented and ren- E. Stelzer), Robert Reinhardt dered data with colors encoding movement directions of the individual Technicians: Claudia Müller, Beate Wittbrodt cells (Keller, et al. 2008). Animal technicians: Aldona Nowicka, Patrycja Grabowska

166 Current Research Gabriel Wittum Computational Neuroscience as part of the Chair Simulation in Technology Detailed Modelling of Sig- nal Processing in Neurons Research Summary Three main aims of contem- Main focus of our work is modeling the signal processing porary brain science with in neurons and networks of neurons in order to obtain a respect to functions of the quantitative understanding of neuronal coding, structure cerebral cortex are a quanti- of cortex at subcellular resolution and plasticity. To that tative understanding of end we develop models, algorithms and software in the framework of cooperative projects. An important 1. Coding of sensory perceptions or motor actions. platform for this cooperation is the newly established Bernstein-Group DMSPiN. 2. Structure of the cortex at subcellular resolution. 3. Plasticity of coding and structure of the cortex. Curriculum Vitae Degrees: 1983 Diploma in Mathematics and Physics, As paradigm for studying these questions we choose University of Kalsruhe a cortical column, the smallest functional unit in 1987 Ph.D. (Dr. rer.nat.) in Applied Mathematics, the mammalian brain. We address these questions University of Karlsruhe with a hierarchical approach starting from spatially 1991 Habilitation, University of Heidelberg resolved models of a single neuron, including detailed 1984-1987: Research assistant, Computer Science, mathematical description of the most relevant physico- University of Kiel chemical processes via local circuits up to ensembles of 1987-1991: Postdoctoral fellow at SFB 123,Univ. of Heidelberg 1991: Professor for Numerics, University of Heidelberg several thousands of neurons in the whole cortical column. 1994-1998: Chair for Applied Mathematics, Head of the Inst. These models are developed in close co-operation with for Computer Applications, University of Stuttgart experiments. Detailed mathematical simulations based 1995: Chairman of the GAMM committee on first principles of physics will lead to novel effective “Scientific Computing” descriptions of neurons and ensembles of neurons. 1996: Co-founder of the SFB 412 These effective descriptions allow the simulation of the 1999-2000: Dean of Mathematics and Computer Science, behaviour of large collections of neurons by advanced University of Heidelberg numerical simulation techniques. The results of the 1996-2006: Head of Competence Center for HLRS different approaches will be compared with each other since 1998: Chair for Simulation in Technology, University of Heidelberg and with data from especially designed experiments for validation. Contact Department of Simulation in Technology To that end, we have to accomplish several sub-tasks: University of Heidelberg Im Neuenheimer Feld 368 First, we need to extract geometries and connectivities of 69120 Heidelberg, Germany neurons from two-photon microscopy data, used to scan A +49-6221-548855 cells inside the brain of a living animal. To that end, we E +49-6221-548860 developed two software tools. The Neuron Reconstruction K [email protected] Algorithm, NeuRA, is a tool for the automatic extraction of o http://sit.uni-hd.de

167 Gabriel Wittum

neuron geometries from two-photon data, [1]. Based on this approach, the following Projects are run in the framework of our newly estabilshed Bernstein- Group “Detailed Modeling of Signal Processing in Neurons (DMSPiN)”. 1. Calcium signaling from synapse to nucleus (with H. Bading) 2. Realistic modeling of presynaptic transmitter content and dynamics and its implications for synaptic plasticity (with A. Draguhn) 3. Development and function of basic networks (with C. Schuster) 4. Modelling of gap junction coupled interneurones (with H. Monyer) The projects are on the level of a single cell. On the level of larger ensembles of neurons, we developed an approach based on compartmental models. Here the key are the two programs NeuGen, generating a network of realistic Fig. 2: Right: Simulation of signal transduction on a branch of a dendrite, neurons, and NeuSim, simulating compartmental models C. Vossen, SiT. on this network on parallel computers.

Fig. 1: Left: Spiny stellate from layer four of a rat’s cortex. Data by J. Waters (NW University, formerly MPI Heidelberg), reconstruction by A. Heusel (SiT, Univ. Heidelberg).

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Top publications • Nägele S. et al. (2003). Large-Eddy Simulation and Multigrid Methods. Electr. Transact. on Num. Analys. 15, 152-164. • Broser, P. J., et al. (2004) Nonlinear anisotropic diffusion filtering of three-dimensional image data from 2-photon microscopy. J. Biomed. Optics 9, 1253-1264. • Eberhard, J., et al. (2004). Coarse graining and multigrid for flow in heterogeneous porous media. SIAM J. Multiscale Mo- delling. • Lang, S., et al. (2005). Large scale density driven flow simulations using parallel unstructured grid adaptation and local multigrid methods. Concurrency Computat. • Eberhard, J. P., et al. (2006). NeuGen: A tool for the generation of realistic morphology of cortical neurons and neural networks in 3D. Neurocomput. 70, 327-342. • Feuchter, D., et al. (2006). A geometry model for the simulation of drug diffusion through stratum corneum. Comput. Vis. Sci 9, 117-130. • Voßen, C., et al. (2007). Modeling and simulation for three- dimensional signal propagation in passive dendrites. Comput. Vis. Sci. 10, 2. • Queisser, G. (2008). Filtering, reconstruction and measurement of the geometry of neuron cell nuclei based on confocal microscopy data. Journal of Biomedical Optics. 13, 014009 (2008); DOI:10.1117/1.2829773.

Structure of the Group Group Leader: Gabriel Wittum Senior Scientists: Michael Heisig Postdoctoral fellows: Vadym Aizinger, Alfio Grillo, Markus Knodel, Michael Lampe, Dimitrij Logaschenko

169 Veit Witzemann Current Research Molecular anatomy of the developing neuro- AChR-mediated activity muscular junction determines early steps in synapse formation Research Summary During early postnatal de- The formation of the neuromuscular junction involves velopment fetal-type AChR reciprocal interactions between the presynaptic nerve (AChRγ) become replaced terminal and the postsynaptic muscle fiber. We generate by adult-type AChR (AChRε). genetically manipulated mouse models to modulate The physiological signifi- synaptic activity by changing the functional properties cance of this conversion is of the postsynaptic signaltransducers, the acetylcholine not understood. We generated by homologous recombi- receptors (AChR) or by inactivating conditionally other nation mice that express during embryonic development synaptic regulators. These studies lead to the analysis of receptors (AChRγ/ε) that have adult-type properties (Koe- disorders that are caused by defective neuromuscular nen et al., 2005). We showed that AChRγ are not required junctions. to preserve myoblastfusion, muscle and end-plate diffe- Curriculum Vitae rentiation since mice that express the modified, adult-type AChRγ/ε during embryonic development are viable and Degrees: 1971 Diploma in Biology, University of Freiburg display no obviously changed phenotype. However, the 1974 Ph.D. University of Constance positioning of endplates is changed dramatically (Fig. 1): 1988 Habilitation in Biochemistry, University of Constance Endplates in mutant mice are spread over a much broader 2008 Apl Professor, University of Constance territory than in wildtype mice suggesting that one func- 1974-1975: Postdoctoral fellow, University of Constance tion of the fetal type of AChR is to ensure an orderly inner- 1975-1978: Research fellow, California Institute of Technology, vation pattern of skeletal muscle. Pasadena, USA 1978-1987: Research associate, Dept. of Neurochemistry, MPI Adult-type AChRε are required to maintain and stabilize for Biophysical Chemistry, Göttingen the postsynaptic architecture (Schwarz et al., 2000). 1987-1989: Group Leader, Dept. of Cellphysiology, MPI for This has been demonstrated by generating mice Biophysical Chemistry, Göttingen that express genetically engineered fetal-type AChR 1989: Group leader, Dept. of Cellphysiology, MPI for Medical Research, Heidelberg throughout postnatal development. The AChRs with “fetal”-like properties induce a “de-differentiation” of the Contact postsynaptic architecture and cause mild symptoms of muscle weakness, indicating that ultrastructure and Department of Cellphysiology MPI for Medical Research function of neuromuscular junctions in adult muscle Jahnstr. 29 depend specifically on the expression of adult-type AChR 69120 Heidelberg (Witzemann et al., in preparation). Germany A +49-6221-486475 In order to understand the regulatory role of AChR- E +49-6221-486459 mediated activity during embryonic development we K [email protected] generated mice that express structurally intact but o http://www.mpimf-heidelberg.mpg.de/forschungsgrup- functionally „silent“ AChR. As described in Peter et al. (2005), pen/witzemannKoenen/index.html we used a mutation located in the subunit gene that was

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We demonstrate that muscle-specific programs of receptor subunit gene transcription control AChR replacement. Analysis of the dynamics of the embryonic AChRγ to adult AChRε conversion at the neuromuscular junction shows, in contrast to previous reports, that conversion does not occur simultaneously for all endplates of the diaphragm muscle. Furthermore, we were able to visualize for the first time channel conversion at distinct endplates. The conversion proceeds in a directed manner in which new AChRε are integrated from peripheral to central regions for individual endplates (Yampolsky et al., 2008).

Synaptic regulators for initial steps in synapse formation and final differentiation of the NMJ We showed that MuSK kinase activity is sufficient to determine early postsynaptic specialization prior to innervation (Sander et al., 2001). We also showed that MuSK kinase is essential for the continuous supply of AChR to the postsynaptic membrane. Conditional inactivation of MuSK in adult muscle causes the disintegration of synapses and induces extensive axonal growth and sprouting. This indicates that both MuSK and AChR maintain the functional and structural integrity of the NMJ (Hesser et al., 2006).

Inherited Channelopathies Congenital myasthenic syndromes (CMS) represent a group Fig. 1: Scattering of endplate locations in diaphragm muscle. Segments of diaphragm compared in adult animals. Locations of endplates in dia- of disorders that are caused by inborn pre- or postsynaptic phragm were measured in muscle stained for postsynaptic acetylcholi- defects of neuromuscular transmission. nesterase activity. In wild type, endplates form a narrow band. In AChRγ/ε The genetic heterogeneity of mutations in the AChR mice, expressing adult-type AChR during embryonic development end- subunit genes leads to variable AChR deficiency and a plates are spread over a broad muscle territory. variable degree of muscle weakness. The molecular basis of postsynaptic CMS is analysed in mouse models lacking found to cause a fast channel congenital myasthenic AChR or expressing AChR carrying mutations that have syndrome in human patients. Preliminary results show been detected in human patients suffering from muscle that the εP121L subunit is assembled into AChR complexes weakness. A new heteroallelic mutation has been detected that are targeted to the postsynaptic membrane. Further in the muscle-specific kinase (MuSK) of a human patient. analysis will clarify whether AChR-mediated activity is We generated a mouse model carrying the same mutation. necessary for the formation and stabilization of synaptic The mice develop severe symptoms of muscle weakness contacts during embryonic development. (Fig. 2) similar to human patients. Thus they represent not only a valuable tool to dissect MuSK signaling pathways In another mouse model we express GFP-labelled γ subunits but also to study MuSK-induced pathophysiology of CMS and thus green fluorescent embryonic-type AChRγ-GFP. (Chevessier et al.,2008).

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Top publications • Rathenberg, J., et al. (2003). High-efficiency transfection of individual neurons using modified electrophysiology techniques. J. Neurosci. Meth. 126/1, 91-98. • Jin, T.-E., et al. (2004). Fiber types of the intrinsic whisker muscle and whisking behavior. J. Neurosci., 24, 3386-3393. • Peter, P., et al. (2005). Mutation of single murine acetylcholine receptor subunits reveals differential contribution of P121 to acetylcholine binding and to channel opening. Pflug. Arch. Eur. J. Physiol. 450, 178-184. • Koenen, M, et al. (2005). AChR channel subtype directs the innervation pattern of skeletal muscle. EMBO Rep. 6, 570-576. • Hesser A.B., et al. (2006). Synapse disassembly and formation of new synapses in postnatal muscle upon conditional inactivation of MuSK. Mol. Cell. Neurosci. 31, 470-480. • Witzemann, V. (2006). The formation of neuromuscular junctions. Cell Tissue Res, 326, 263–271. • Yampolsky P., et al. (2008). AChR channel conversion and AChR-adjusted neuronal survival during embryonic development. Mol. Cell. Neurosci. 37, 634-645. • Jin T.-E., et al. (2008). Changes in AChR function induce shifts in muscle fiber type composition. FEBS Journal 275, 2042–2054. • Teichert, R. W., et al. (2008). Peptide-toxin tools for probing the expression and function of fetal and adult subtypes of the nicotinic acetylcholine receptor. Ann. N.Y. Acad. Sci. 1132, 61- 70. • Chevessier, F., et al. (2008). Mutation in MuSK leads to haploinsufficiency in mice inducing a muscle weakness similar to human congenital myasthenic syndromes. Hum. Mol. Gen., 17, Fig. 2: Heteroallelic mutation in musk causes CMS. The transgenic mice 3577–3595 will help to understand MuSK-induced pathophysiology of CMS at the molecular level. Mouse mutant at P 40 and P 75 displays progressive shrinkage of pelvic and scapular muscles and appearance of severe thoracolumbar kyphosis.

With our mouse models we can investigate in living animals experimentally induced and clearly defined perturbations of signal transduction at an as yet unknown resolution. Biochemical, ultrastructural, and electrophysiological changes can be followed in vivo. We employ direct gene transfer to modulate signal cascades, initiated by AChR-mediated activity or by MuSK-induced signals Structure of the Group and quantitive PCR, and siRNA technology to analyse Group Leader: Veit Witzemann changes in gene expression. Finally, we visualize synapse Postdoctoral fellows: Frédéric Chevessier, Pessah Yampolsky degradation and synapse formation by in vivo imaging of PhD-Students: Pier-Giorgio Pacifici individual neuromuscular junctions. Technical Assistance: Ulrike Mersdorf, Susi Zobeley

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173 Teaching

IZN Teaching Program • Prof. Dr. Stephan Frings (Institute for Zoology) • Prof. Dr. Joachim Kirsch (Institute for Anatomy and Cell Biology) The Major ‘Neuroscience’ in the MSc program ‘Molecular • Prof. Dr. Thomas Kuner (Institute for Anatomy and Cell Biology) Biosciences’ • Dr. Frank Möhrlen (Institute for Zoology) • Prof. Dr. Elisabeth Pollerberg (Institute for Zoology) To prepare students for PhD programs in neuroscience, the IZN • Prof. Dr. Christoph Schuster (Institute for Neurobiology) offers a neuroscience course as part of the MSc program ‘Mole- • Prof. Dr. Rainer Spanagel (Central Institute of Mental Health) cular Biosciences’ at the Faculty of Biosciences. After finishing • Dr. Rolf Sprengel (Max-Planck-Institute for Medical Research) the BSc, students may enrol in this 2-year course and receive • Prof. Dr. Klaus Unsicker(Institute for Anatomy and intensive training in neuroscientific theory and cutting-edge Cell Biology) lab technology. The program is taught in English and welcomes international students. With a strong emphasis on experimental The program launched in 2007 is designed for twelve highly approaches, the program’s aim is to prepare students for a career motivated and talented students each year. in the neurosciences. The MSc can be completed in either 3 or 4 semesters. IZN PhD Program The program: 1st semester: Established in April 2004, the doctoral program at the Interdis- • Frontiers of Biosciences I + II (Lectures in molecular biosciences, ciplinary Center for Neurosciences leads to the degree of Dr. rer. including molecular neurobiology) nat. (in the Faculty of Biological Sciences) or of Dr. med. (in the • Lab class ‘Modern methods in neurobiology’ with seminar Faculty of Medicine). • Lab class ‘Axon growth, synaptogenesis, and synaptic plasticity’ with seminar The program provides research opportunities in the fields of Neuroanatomy, Neurobiology, Clinical Neurobiology, Neuro- 2nd semester: physiology, and Medical Cell Biology. With the new structure of • Focus Bioscience I: ‘Neuronal structures and molecules’ the IZN additional topics of Neuroscience (Behaviour, Modelling, (Lectures, lab rotation and seminar) Neurodegenerative Diseases, Pain, Psychiatric Disorders etc.) will • Focus Bioscience II: ‘Neuronal networks and computation’ be covered. (Lectures, lab rotation, seminar) It also includes doctoral students from the Graduate College 3rd semester: 791 ‘Neural Developmental and Degenerative Processes: Basic • Biolab: Neuroscience lab rotation program Research and Clinical Implications’ which is currently in its second • Working in Biosciences: Lab rotation program of the student’s funding period. choice Presently, the program comprises 45 doctoral students, 17 of whom are funded by the Graduate College 791, with the remain- 4th semester: der receiving support from various stipends and institutional • Master’s thesis in a neuroscience lab. funds. Instruction covers a wide spectrum of neuroscientific research, ranging from molecular neurobiology to brain physiology as well The program provides students with a variety of lectures and as cognitive and computational neuroscience. The MSc program seminars, as well as with the possibility to participate in several is organized by the following IZN neurosciences lecturers: of the following practical courses offered by members of the IZN.

• Prof. Dr. Hilmar Bading (Institute for Neurobiology) • Preparation and culture of primary neurons (H. Bading) • Dr. Peter Bengtson (Institute for Neurobiology) • Video-imaging of neuron cultures (H. Bading) • Dr. Francesca Ciccolini (Institute for Neurobiology) • Electrophysiology (H. Bading) • Prof. Dr. Andreas Draguhn (Institute for Physiology • Transmission electron microscopy (H. Bading) and Pathophysiology)

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Pre- and Postgraduate Teaching Programme

Bachelor Biology Bachelor Molecular Cellbiology Seminars, Practical Courses, Lectures

Master Molecular Biosciences Major Neurosciences Seminars, Practical Courses, Lectures

HBIGS Selection, Admission, Administration

SFB 636 Graduate IZN PhD Programme Graduate College 791 Programme

Medical Faculty Faculty of Bioscience MD-PhD Programme of the Medical Faculty Dr. med. Dr. rer. nat. and the Faculty of Biosciences

175 Teaching

• Multiple electrode arrays (H. Bading) With its new structure, the IZN integrates 51 IZN Investigators and • Confocal laser scanning microscopy (H. Bading) thus provides a context in which students in the PhD program • Isolation and in vitro culture of neural stem cells (F. Ciccolini) will have extensive opportunities to enhance their knowledge in • Fluorocytometry (F. Ciccolini) the fields of behavioral/systems and translational neurosciences. • Associative learning and memory (C. Schuster) • Drosophila molecular and classical genetics (C. Schuster) • Electrophysiology – Intracellular current clamp recordings (C. Schuster) • Functional Imaging – calcium and FM 1-43 (C. Schuster) • In vitro and in vivo electrophysiological recording in the hippocampus (H. Monyer) • Paired recordings from connected neurons in brain slices (A. Rozov) • Preparation and culture of primary neurons (J. Kirsch) • Video-imaging of neuron cultures (J. Kirsch) • Transmission electron microscopy (J. Kirsch) • Confocal laser scanning microscopy (J. Kirsch) • Transmission electron microscopy (K. Unsicker) • Neuronal cell culture (K. Unsicker) • Non-radioactive in situ hybridisation on tissue slices (U. Ernsberger) • Visualisation of the developing mouse nervous system by GFP (K. Tucker) • Transfection, imaging, and morphological analysis of primary neuronal cultures (G. Schratt) • Immunohistochemistry on floating brain sections (H. Simon) • Fluorescence-imaging and patch clamp electrophysiology in cultured neurons (U. Misgeld and A. Draguhn) • Patch clamp recording from identified neurons in brain slices (U. Misgeld and A. Draguhn) • Measurement and analysis of neural network oscillations (U. Misgeld and A. Draguhn)

At the IZN’s annual retreat, PhD students have the chance to present a poster to the assembled members of the center and invi- ted symposium speakers.

Several IZN Investigators are also members of ‘The Hartmut Hoff- mann-Berling International Graduate School of Molecular and Cellular Biology Heidelberg’ (HBIGS) which is part of the ‘Exzel- lenzinitiative’ approved in October 2007.

176 Diploma Theses

Diploma Theses 2003-2008 tifying potassium channels Oehmen, Martin (2008) Bading Effects of presynaptic GABA-A-receptors on inhibitory synaptic Weislogel, Jan (2003) transmission in cultured hippocampal neurons Calcium imaging using recombinant Ca2+ probes in primary Kameke, Alexandra von (2008) hippocampal neurons Optical imaging of fast network activity Wiegert, Simon (2004) (I) Live Imaging of ERK Nuclear Translocation and (II) Analysis Flor of the Role of Gene Transcription in Activity Induced Enhance- Caumans, Indra (2003) ment of Synaptic Efficacy in Hippocampal Neurons in Culture Dysfunktionale Aufmerksamkeitsprozesse und Präpulsinhibiti- Hoffmann, Tina (2006) on bei schizophren erkrankten Menschen Nuclear Translocation of the CREB Coactivator TORC2 (Trans- Cetinkaya, Sevim (2003) ducer of Regulated CREB Activity 2) and Examination of CREB Das Schreckreflexparadigma: Emotionale Verarbeitung visuel- mediated Activity-Dependent Neuronal Survival in Cultured ler Reize und reizinduziertes Verlangen in Verbindung mit den Hippocampal Neurons Persönlichkeitsmerkmalen “Schadensvermeidung” und “Neu- Krieger, Markus (2007) gierverhalten” bei abstinenten Alkoholikern. Effects of nuclear Ca2+ and CaMKIV signalling on Nucleus Ac- Lamberger, Michael (2003) cumbens-gated Behavior in the Rat Das Schreckreflexparadigma: Emotionale Verarbeitung visuel- Vicinus, Benjamin (2007) ler Reize und reizinduziertes Verlangen in Verbindung mit den Interfering with nuclear ERK1/2-signaling in cultured hippo- Persönlichkeitsmerkmalen “Schadensvermeidung” und “Neu- campal neurons gierverhalten” bei abstinenten Alkoholikern. Ditzel, Désirée (2008) Martin, Veronika (2003) Imaging of spacially distinct calcium signal using recombinant Evaluation eines Reizexpositionsverfahrens zur Behandlung calcium probes alkoholabhängiger Patienten Schlüter, Jana (2008) Rumpf, Matthias (2003) Calcium imaging in Hydra vulgaris Die Rolle von Lernprozessen und elterlichem Verhalten bei der Entwicklung von sozialen Ängsten Tröster, Philip (2008) Calcium signaling and activity-dependent gene expression in Someasan, Alexandra (2003) hippocampal neurons Die P50-Suppression schizophren Erkrankter erhoben in einer simultanen P50-PPI-Abteilung Bartsch Stroth, Sanna (2003) Panke, Jutta (2007) Somatosensorisch evozierte Potentiale bei chronischen Rho and MAPK signal transduction pathways in the MEGAP Schmerzen der Skelettmuskulatur knockout mouse Wald, Annette (2003) Ciccolini Untersuchung des Zusammenhangs der Inhibitory Gating-Ma- ße Präpulsinhibition und P50-Suppression an gesunden und Hundeshagen, Phillip (2007) schizophrenen Probanden Effect of Neural Activity on the Differentiation of Neural Stem Cells Chaney, Vernon (2004) Untersuchungen zur Wahrnehmung und zum Erkennen emoti- Draguhn onaler Gesichtsausdrücke bei der sozialen Phobie Lehmann, Alexander (2007) Dos Santos, Vasco (2004) Actions of Aß-peptides on synaptic transmission Zur Spezifität der Emotionsinduktion bei Patienten mit genera- Duhme, Nana (2008) lisierter Sozialphobie und nicht ängstlichen Kontrollpersonen Characterisation of normal and mutant human inwardly rec- in einem Verhaltenstest

177 Diploma Theses

Nees, Frauke (2004) Ullrich-Kleinmanns, Jens (2005) Psychophysiologische Untersuchungen bei alkohol-/nikotin- Neuropsychologische Aspekte der Nikotinabhängigkeit: Zur abhängigen Patienten - Analyse der Reaktion auf verschiedene Veränderung der motivationalen Valenz verschiedener Verstär- Alkohol- bzw. Nikotinreize ker im Nikotinentzug Pfeiffer, Kerstin (2004) Farrugia, Claire (2006) Traumaberichte von Personen mit und ohne PBS: inhaltliche Quantitative sensory testing and psychological variables in tri- Textanalyse sowie subjektive und psychophysiologische Reak- geminal neuralgia, trigeminal neuropathy and atypical facial tionen pain Scharpf, Katrin (2004) Flach, Florentine (2006) Validierung einer Druckstimulationsmethode zur experimen- Der Einfluss von Persönlichkeitsmerkmalen auf die Posttrau- tellen Schmerzinduktion matische Belastungsstörung Storner, Tina (2004) Hoffmann, Mike (2006) Psychophysiologische Untersuchungen bei alkohol-/nikotin- Wiederauffrischung (reinstatement) - ein Mechanismus zur abhängikgen Patienten - Analyse der Reaktion auf verschiede- Aufrechterhaltung der sozialen Phobie ne Alkohol- bzw. Nikotinhinweisreize Kehrberger, Christiane (2006) Tuttas, Marie-Luise (2004) Schmerzverarbeitung und Schmerzsensibilität bei Kindern mit Psychometrische Evalution von Fragebögen zur Schmerzdiag- rezidivierenden Bauchschmerzen nostik von Kindern und Jugendlichen Koler, Johanna (2006) Vogl, Stephan (2004) Der Einfluss von Delta-9-THC auf Lernen und Gedächtnis Zur Rolle der Antizipation bei der stressbedingten Aktivierung Konieczna, Anna (2006) der Hypothalamus-Hypophysen-Nebennierenrinden-Achse Mehrdimensionalität psychophysikalischer Kennwerte experi- Balke, Doreen (2005) menteller Schmerzverarbeitung bei Gesunden und chronisch Langzeitauswirkungen früher Schmerzerfahrungen auf die Schmerzkranken Schmerzwahrnehmung in später Kindheit Krajewski, Robert (2006) Dilmac, Kristil (2005) Können endokrinologische Parameter anhand von Stressver- Soziale Kompetenz bei Subtypen sozialer Phobie arbeitungsstrategien, alltäglicher Belastung, Depressivität und Halbeis, Damaris (2005) Ängstlichkeit vorhergesagt werden? Klassifikation chronischer Gesichtsschmerzen anhand des MPI-D Lang, Stefan (2006) Herrmann, Sandra (2005) Zum Einfluss stresskorrelierter Muskelanspannung bei Patien- Psychometrische Evaluation von Fragebögen zur Diagnostik ten mit Trigeminusneuralgie bei Schmerzen und allgemeinen körperlichen Beschwerden Rance, Mariela (2006) im Kindes- und Jugendalter Sensibilisierung bei verschiedenen Stimulationsmethoden Heuser, Mark (2005) Suppl Angelika (2006) Emotionale Reagibilität bei generalisierter sozialer Phobie Mögliche Einflussfaktoren für die Entstehung einer sozialen Mußgnug, Nadga (2005) Phobie Angstinduktion mittels phobierelevanten und -irrelevanten Yimaz, Pinar (2006) Stimuli bei nicht-gerneralisierter sozialer Phobie Zusammenang zwischen quantitativer sensorischer Testung Schreiber, Viola (2005) und der Beeinträchtigung bei Patienten mit chronischen mus- Emotionale Taubheit in der Posttraumatischen Belastungsstö- kuloskelettalen Schmerzen rung Zahran, Assad (2006) Soekadar, Surjo (2005) Der Einfluss von Lernerfahrungen auf implizite Assoziationen Phantomschmerz und neuronale Plastizität nach Amputation bei Personen mit sozialer Phobie und nichtängstlichen Kont- der unteren Extremität: Prävention und Therapie mit Meman- rollpersonen tine und Baclofen

178 Diploma Theses

Klimm, Sabrina (2006) Vocke, Kerstin (2006) Das Schmerzerleben von Kindern mit rezidivierenden Bauch- Mutagenese der cAMP-Bindestelle von CNG-Kanälen schmerzen: eine experimentelle Untersuchung unter Verwen- Hengl, Thomas (2006) dung des Kaltwassertests Lokalisation von mRNA-Transkripten auf Gewebeschnitten des Greiner, Stefanie (2007) Riechepithels Die Rolle der Mutter-Kind-Interaktion während des Kaltwasser- Schlabing, Jochen (2006) tests bei Kindern mit rezidivirenden Bauchschmerzen Expression von Calmodulinmutanten Lieven, Stefan (2007) Stavermann, Maren (2007) Wenn die Angst, vor anderen das Gesicht zu verlieren, Men- Fluoreszenzoptische Messungen der Riechzellaktivität schen aus der Bahn ihres Alltags wirft Jeridi, Semir (2007) Wittenberg, Sandra (2007) Etablierung eines fluoreszenzbasierten Nachweissystems für Gibt es Geschlechtsunterschiede in der Schmerzwahrneh- Ca2+-aktivierte Cl-- Flüsse mung von Kindern mit Migräne Daiber, Philipp (2007) Dinu-Biringer, Ramona (2008) Expression von Bestrophin 2 in olfaktorischen und nozizepti- Elektrodermale Reaktivität und neuronale Korrelate beim aver- ven Systemen siven Lernen und Verlernen. Vergleich von Cue- und Kontext- Konditionierung im Zusammenhang mit Ängstlichkeit Holstein Thiel, Kathrin (2008) Bielen, Holger (2003) Aufmerksamkeitshinwendung auf alkoholassoziierte Stimuli: Identifizierung der Interaktionspartner des T-Box Transkripti- Zusammenhang mit der kognitiven Leistungsfähigkeit und onsfaktors Brachyury in Hydra der Inhibition präpotenter Verhaltensreaktionen bei alkoho- Philipp, Isabelle (2003) labhängigen Personen Cloning and Expression of Cell Polarity Genes in Hydra Frings Saina, Michael (2003) Savic, Alexandra (2004) Isolation und Expressionsanalyse von Mitgliedern eines TGF- Suche nach Ionenkanalblockern für CNG-Kanäle in Schlangen- beta Signalwegs in Nematostella vectensis und Kegelschneckengiften Vocke, Dirk (2003) Drexel, Jan (2004) Funktionale Charakterisierung von Hydra Chordin zur Vorberei- Isolierung und Charakterisierung einer Astacin-homologen tung von Injektionsexperimenten und Protein-Interaktionsstu- Metalloprotease dien mittels Blau-Nativer PAGE Mayer, Ulrich (2004) Anton, Roman (2004) Immunohistochemische Charakterisierung von Schmerzzellen Analyse eines BMP2/4 Dpp-Orthologs und eines potentiellen in der Ratte BMP-Antagonisten von Nematostella vectensis (Anthozoa, Ed- wardsiidae) Wiedmann, Verena (2004) Internetportal der Bioinformatik für Studenten im Grundstudi- Fritzenwanker, Jens (2004) um der Biologie Beschreibung der Embryonalentwicklung und Charakterisie- rung eines Fork Head Homologs aus Nematostella vectensis Klimmeck, Daniel (2005) Vitale Gewebeschnitte des olfaktorischen Epithels der postna- Hrach, Jens (2004) talen Maus Mikroinjektionen in Nematostella vectensis Ungerer, Nicole (2005) Mättner, Robert (2004) Isolierung und Charakterisierung von Membranproteinen aus Analyse der Transfektion von Hydrazellen mit Transposon ent- Riechzellen der Ratte haltenden Vektoren Funk, Katharina (2006) Nacak, Tanju (2004) Entwicklung eines Modellsystems für die Signaltransduktion in Charakterisierung von Signalmolekülen während der Kopfre- Schmerzzellen generation in Hydra vulgaris

179 Diploma Theses

Büttner, Andreas (2005) Kirsch Analyse putativer BMP- und Wnt-Anatgonisten mit einem Cys- Laue, Thomas (2007) tin-Knoten Motiv in Hydra Lokalisationsstudien des eukaryotischen Elongationsfaktors Krause, AnnKatrin (2006) (eEF1 A) in hippocampalen Neuronen und Etablierung von Charakterisierung von Gremlin-ähnlichen Molekülen in Hydra RNA-Interferenz-Methoden zur Reduktion der Expression von magnipapillata eEF1 A Steinbronn, Nadine (2006) Kuner Thomas Cadherine in Nematostella vectensis - genauere Charakterisie- rung und Interaktionstudien mit beta-Catenin. Vasileva, Mariya (2005) Overexpression of synapsin isoforms in the calyx of Held via Veit, Marion (2006) ATGp Charakterisierung eines PS-1 und eines APP Homologs in Hyd- ra magnipapillata Reuter,Kirsten (2006) Selective Labeling of Presynaptic Protein in the Calyx of Held Christ, Annabel (2007) Mediated by the Genetically Encoded AGT-tag ntersuchungen zur stabilen Transfektion von Hydra magnipapillata Sandikci, Arzu (2006) Imaging steady state spatial distribution of chloride in the Kins soma and dendrites of principal neurons using new variants of Nesic, Iva (2002) genetically encoded chloride indicator Clomeleon Untersuchungen zur subzellulären Lokalisation von APLP1 und Monyer APLP2 Schächinger, Thorsten (2003) Haas, Petra (2003) Generation of a DNA construct for the conditional knock-out of Charakterisierung subzellulärer Sortierungssignale von APP the NMDA receptor subunit NR2B gene in Mus musculus Szodorai, Anita (2003) Götz, Thomas (2004) Untersuchungen zur Funktion der APP-Proteinfamilie als Kine- Developing a novel method to reversibly modulate neuronal sin-Membranrezeptoren activity in the mouse brain Wagner, Katja (2004) Bocklisch, Christina (2007) Untersuchungen zur Interaktion der APP-Genfamilie mit PSD- Functional and neurochemical characterization of NR2D-EGFP 95 und Fe65/Fe65L1 expressing cells in the hippocampus Back, Simone (2005) Untersuchungen zur subzellulären Verteilung von APP in Ner- Pollerberg venzellen Jungbauer, Simon (2003) Lauther, Nadine (2005) Einfluss von mikro- und nanostrukturierten Oberflächen auf Untersuchungen zur axonalen Transportmaschinerie von APP Melanozyten und Osteoblasten Familienmitgliedern Tchouandong, Leopoldine (2003) Siehl, Katjuscha (2005) Produktion und Aufreinigung der rekombinanten Zelladhäsi- RNA Interference Technology Based Knockdown of the APP onsmoleküle DM.GRASP u. ALCAM Familiy Proteins Bezler, Martin (2004) Rusu, Patricia (2005) Einfluss unterschiedlich nanostrukturierter Oberflächen auf Analyses of the Physiological Functions of APP/APPL in Droso- das Verhalten von Osteoblasten phila melanogaster Blüm, Raphael (2004) Stahl, Ronny (2007) Durchmusterung einer cDNA-Bank nach extrazellulären Inter- The influence of APP processing on cell adhesion aktionspartnern von Zelladhäsionsmolekülen Souren, Marcel (2004) In vivo analysis of regulatory elements predicted in silico

180 Diploma Theses

Georg, Tanja (2005) Laudenklos, Sabrina (2004) Regulation der Endozytose von Zelladhäsionsmolekülen Lentiviral-based approach to study dendritic integrative pro- Krais, Annette (2005) perties of pyramidal neurons in rat barrel cortex in vivo Einfluss des Zelladhäsionsmoleküls DM-GRASP auf intrazellu- Spors läre Signalwege Shahshahani, Nilufar (2008) Bertuch, Stefanie (2006) Odor adaptation and odor discrimination time Regulation der Endozytose des Zelladhäsionsmoleküls DM- GRASP Sprengel Bubis, Andreas (2007) Layer, Liliana (2003) Untersuchungen zur Interaktion von Zelladhäsionsmolekülen Funktioneller Austausch der endogenen GluR-A-Untereinheit Geuder, Tanja (2007) durch GluR-A-Varianten im Hippokampus der Maus Untersuchungen zur Interaktion zweier integraler, axonaler Marx, Verena (2003) Membranproteine Tetrazyklinregulation GFP markierter AMPA-Rezeptoren im Ge- Höckendorf, Burkhardt (2007) hirnpräparat transgener Mäuse Proof of principle for a gene trap approach in medaka using Mihaljevic, André (2004) the maize transposable element Ac/Dc mRNA trafficking of the voltage - gated potassium channel subunit Kv4.2 in hippocampal pyramidal neurons. Rupp Bus, Thorsten (2005) Andermann, Martin Selektive Steuerung der Genexpression in GnRH-Neuronen. Neuromagnetic investigation of instrument size: Influence of musical aptitude and pitch perception Tang, Wannan (2005) In vivo labelling of Tbr1 gene in mitral cells in the olfactory bulb Ibrom, Sophie of developing zebrafish, Danio rerio Mismatch negativity, the auditory memory trace and musical aptitude Strobel, Cornelia (2006) Analyse des Isolator 40 Elements in transgenen Mäusen Schuster Tucker Pfenning, Philipp-Niclas (2007) Die Rolle von TRP-Calciumkanälen und NO-Signalkaskaden Brachmann, Isabel (2005) während erfahrungsabhängiger Potenzierung glutamaterger Analysis of the central and peripheral nervous systems of the Synapsen mouse mutant line tauGFP during embryonic development Leefmann, Jon (2008) Schumacher, Stefanie (2006) Analysis of synaptic transmission and locomotive behavior in Establishment of RNA interference (RNAi) for the reduction of larvae of Drosophila melanogaster by optical control of phosen- gene expression in neural stem cells from mouse brain and sitive adenylyl cyclase and channelrhodopsin-2 from in vitro-differentiated embryonic stem cells Litzenburger, Ulrike (2007) Weißmüller, Kathrin (2006) Die Rolle des Phosphorregulierungsstaus der postsynaptischen The neurobiological developmental potential of in vitro-diffe- Glutamatrezeptoren und die Rolle der NO-Synthese während rentiated embryonic mouse stem cells in the central and peri- der erfahrungsabhängigen synaptischen Potenzierung von pheral nervous system of the chick embryo Drosophila melanogaster Shen, Xiao (2008) Analysis of a cortical layering defect in the postnatal brain of Seeburg tau::N19RhoA transgenic mice Cetin, Ali Haydar (2004) Lentivirus-mediated gene expression in the mammalian cor- Unsicker tex Lohr, Jennifer (2004) Die Entwicklung der Nebennieren und sympathoadrenalen Zelllinie bei SF-1 haploinsuffizienten Mäusen

181 Master / Doctoral Theses

Master Theses 2003-2008 Doctoral Theses 2003-2008 Bading Bading Aso, Yoshinor (2007) Dübel, Jens (2005) Activity Dependent Induction of GADD45 Family Genes and Untersuchung intrazellulärer Chlorid-Konzentrationen in Clo- Promotion of Survival in Hippocampal Neurons meleon-exprimierenden ON-Bipolarzellen der Mausretina mit Burau, Karin (2007) Hilfe der Zwei-Photonen-Mikroskopie The morphology and dynamics of the retinal bipolar cell termi- Inta, Joana Monica (2005) nals in the zebrafish TWEAK and NF-kB in Cerebral Ischemia Lu, Li (2007) Bucher, Julia (2006) Characterization of Activity-regulated Fbxo33, a Potential Gene Early development of topographically organized activity pat- Involved in Synaptic Plasticity terns and GABAergic Interneurons in the zebrafish olfactory Zou, Ming (2008) bulb A Nuclear Calcium-Regulated Gene Program for Neuroprotec- Chen, Jing (2006) tion Cerebellar granule cell-specific deletion of the AMPA receptor subunit GluR-D gene Draguhn Hartmann, Bettina (2006) Vasileva, Mariya (2007) Molekulare Mechanismen synaptischer Plastizität in nozizepti- Alterations of synaptic transmission after disruption of the ven Neuronen des Rückenmarks unter Beteilligung von AMPA- APPpathway Rezeptoren Schratt Wittmann, Malte (2006) Khudayberdiev, Sharof (2007) Synaptic and extra-synaptic NMDA receptors in hippocampal The transcriptional regulation of a large microRNA cluster at neurons: regulation of nuclear shape and cell fate the rat 6q32 domain Zhang, Sheng-Jia (2006) Veith, Matthias (2007) Specifying molecular determinants of the subcellular targeting Identification of neuronal microRNA target mRNAs of synaptic and extrasynaptic GABA(A) receptors Eder, Anja (2007) Tucker Generation and analysis of calcium signals in the cell nucleus Isabel Brachmann (2005) Sanno, Hitomi (2007) Analyse des Zentralen und Peripheren Nervensystems der Postnatal control of cortical development by Rho GTPases Mausmutantenlinie tau-GFP während der Embryonalentwick- lung Weislogel, Jan (2008) Imaging of spatially distinct calcium-signals using recombinant Stefanie Schumacher (2006) indicators Etablierung von RNA Interference (RNAi) zur Minderung der Genexpression in neuralen Stammzellen aus dem Mausgehirn Bartsch und von in vitro-differenzierten embryonalen Stammzellen Baur, Max (2006) Kathrin Weissmueller (2006) Charakterisierung tet-kontrollierter, serotonerger oder glialer Das neurobiologische Entwicklungspotential von in vitro-dif- Gehirnexpression des artifiziellen Transkriptionsfaktors (tTA) ferenzierten embryonalen Maus-Stammzellen in zentralen und mittels doppelt transgener Reporter-Mäuse peripheren Nervensystem von Hühnerembryonen Kautt, Sandra (2007) Wolff, Steffen (2007) Expression of MEGP mRNA during embryonic development Nuclear-cytoplasmic shuttling of histone deacetylases and its Mannhardt, Sönke (2007) role in neurogenesis Effect of nifedipine on fear memory extinction

182 Doctoral Theses

Huppert, Verena (2008) Hausselt, Susanne (2006) Charakterisierung tet-kontrollierter Überexpression des Sero- Computation of direction selectivity in retinal starburst amac- tonin-Transporters in serotonergen Neuronen (TPH2-tTA/lacZ- rine cell dendrites − studied using electrophysiological recor- tetO-SERT) dings and two−photon imaging Böhm, Gerald (2008) Breuninger, Tobias (2006) Charakterisierung konditionaler serotonerger Rekombination Chromatic processing in the retina durch das Cre/LoxP-System mittels doppelt transgener Repor- Nanguneri, Siddharth (2008) ter-Mäuse (R26R TPH2-CreERT2) Mechanisms of dendritic processing in retinal amacrine cells Ciccolini Flor Gakhar-Koppole, Nidhi (2006) Koeppe, Caroline (2003) Effect of neural activity on neural precursors differentiation Mechanismen kortikaler Reorganisation nach Beinamputation Carillo-García, Carmen (2008) Peper, Martin (2003) Role of Growth/Differentiation Factor (GDF) 15 in the regulati- Untersuchungen zur neurobehavioralen Toxizität polychlorier- on of embryonic neural precursors ter Kohlenwasserstoffe nach chronischer Exposition im Nied- Draguhn rigdosisbereich Maier, Nicolaus (2004) Stolle, Annette M. (2003) Mechanisms of fast network oscillations in mouse hippocam- Taktile Illusion: Wahrnehmung und neuronale Analyse spatio- pal slices temporaler Reizmuster Hartmann, Kristin (2007) Wessa, Michéle (2003) Homeostatic plasticity of GABAergic synapses in the hippo- Dissociation between explicit and implicit memory in Posttrau- campus matic Stress Disorder Golovka, Tatyana (2007) Diener, Carsten (2004) Effects of cannabinoids on GABA-A-receptors Der Zusammenhang zwischen dem Konditionierungs- und Ar- beitsgedächstnisdefizit von schizophrenen Erkrankten Holter, Nadine (2008) Maturation of inhibitory signalling within the dentate area of Frew, Ashley (2004) the mouse The influence of discouragement, anxiety and anger on pain: An examination of the role of endogenous opioids Voinova, Olga (2008) Developmental changes of release at neocortical synapses Knoblauch, Christine (2004) Soziale Kompetenz und dysfunktionale Kognitionen von Per- Bähner, Florian (2008) sonen mit einer generalisierten Sozialphobie und nicht ängst- Plasticity of CA1 pyramidal neurons in oscillating networks lichen Kontrollpersonen in einem Verhaltenstest Grimm, Christiane (2008) Löber, Sabine (2004) Effects of A-beta peptides on synaptic transmission Exposure therapy in alcohol dependence Weiß, Elisa (2008) Stöckel, Cornelia (2004) Modulation of hippcoampal networks by glucocorticoids Reorganisation des primären somatosensorischen Kortex bei Birke, Gunnar (2008) Thalidomid-Embryopathie Serotonergic modulation of hippocampal network oscillations Welzel, Helga (2004) Euler Neuropsychological and structural predictors of therapy success in alcohol dependence Dübel, Jens (2005) Two-photon imaging reveals somato-dendritic chloride gra- Albrecht, Ulrike (2005) dient in retinal ON-type bipolar cells expressing the biosensor Reizreaktion und Verlangen bei pathologischen Glücksspie- Clomeleon. lern: Psychologische und physiologische Parameter

183 Doctoral Theses

Christmann, Christoph (2005) Leménager, Tagrid (2007) Integration von Elektroenzephalographie (fMRT) bei somato- Operationalisierung differenter Craving Dimensionen und sensorisch evozierter zerebraler Aktivität Validierung an psychologischen sowie neurophysiologischen Damian, Marinella (2005) Kriterien Der Nutzen neuropsychologischer Speed-Tests für die Früh- Ludäscher, Petra (2007) und Differentialdiagnostik dementieller Syndrome und subkli- Somatosensorik und Schmerzwahrnehmung bei Patientinnen nischer Formen kognitiver Beeinträchtigungen mit Borderline-Persönlichkeitsstörung Demirakca, Traute (2005) Niederbühl, Eva Nastassja (2007) Neuronale Aktivierung bei einfachen assoziativen Lernprozes- Schmerzwahrnehmung und kortikale Verarbeitung bei Pa- sen: Darstellung einer einfachen assoziativen Lernaufgabe mit tienten mit chronischer Rückenmarkstimulation: Periphere funktioneller Kernspintomographie und Vergleich zwischen elektrische Stimulation und elektroenzephalografische Unter- gesunden und schizophrenen Patienten suchungen Hofer, Anne K. (2005) Pohlann, Grit Kerstin (2007) Reliabilität und Validität eines Fragebogens zum Screening Differentielle klassische Konditionierung emotionaler Reaktio- von Schlafstörungen nen auf Fazialreize bei generalisierten Sozialphobikern Mc Avinue, Laura (2005) Ridder, Stephanie (2007) The Use of Imagery in Rehabilitation Psychophysiologie und Glucocorticoid-Rezeptor: Hinweise für Diers, Martin (2006) die Entstehung psychischer Erkrankungen The role of illusory hand movements in cortical reorganization Thalemann, Ralf (2007) and phantom phenomena after unilateral arm amputation Variablen exzessiver Computer- und Internetnutzung im Kin- Klossika, Iri (2006) des- und Jugendalter Schmerzsensibilität und –sensibilisierung bei chronischen Weber, Nadja (2007) muskuloskelettalen Schmerzen Plasmatische und serologische Veränderungen von Interleu- Paul, Birgit (2006) kin-6, Tumornekrosefaktor-α und Neuropeptid-Y bei antizi- Psychologie und Pathophysiologie von Patientinnen mit chro- pierten und mittels Cholezystokinin-Tetrapeptid induzierten nischen Rücken/ Kreuzschmerzen nächtlichen Panikattacken Struve, Maren (2006) Zohsel, Kathrin (2007) Emotional processing in tinnitus Schmwerzverarbeitung bei Kindern mit verschiedenen Arten Barth, Johanna (2007) chronischer Schmerzen Subgruppendifferenzierung bei Patienten mit Borderline- Gierhan, Sven (2008) Persönlichkeitsstörung unter Berücksichtigung von dimensio- Die Rolle des Hippocampus bei Posttraumatischen Belastungs- naler Persönlichkeitsausprägung, Komorbiditäten und Symp- störungen - eine fMRT-Studie tomausprägung Sandgathe Husebo, Bettina (2008) Bühler, Mira (2007) Assessment of Pain in Patients with Dementia Functional Imaging of Motivational and Emotional Processing in the Healthy and Addicted Brain Frings Eichler, Miriam (2007) Gilbert, Daniel (2006) Empfindungsveränderungen bei Patienten mit Spinal Cord Chlorid-basierte Signalverstärkung in Capsaicin-sensitiven Atimulation: Ergebnisse quantitativer sensorischer Testung bei Schmerzzellen chronischen neuropathischen Schmerzen in einer unteren Ex- Mayer, Ulrich (2008) tremität Charakterisierung des Proteoms olfaktorischer Cilien Jochims, Anja Marie (2007) Klimmeck, Daniel (2008) Kortikolimbische Verarbeitung von Schmerzreizen und imagi- Molekulare Grundlagen exzitatorischer Cl- Ströme in Riechsin- nierter Selbstverletzung bei der Borderline-Persönlichkeitsstö- neszellen rung

184 Doctoral Theses

Gass Blüm, Raphael (2008) Zörner, Björn (2004) Funktionelle Beziehungen zwischen GlyR-Aktivität und GlyR Verhaltensbiologische und neurochemische Untersuchungen assoziierten Proteinen zur Neurotrophin-Hypothese der Depression mit transgenen Kremer, Thomas (2008) Mäusen Identifications of novel presynaptic proteins Chourbaji, Sabine (2005) Köhr Coping with stress: impact of the hypothalamic pituitary adrenal system and neurotrophic circuits in the learned helpless- Schupp, Bettina (2004) ness modell of depression GluR-B und GluR-Blong: Einfluss auf die synaptische Lokalisati- on und Transmission von AMPA-Rezeptoren im Hippokampus Holstein Pawlak, Verena (2004) Cramer v.d. Laue, Christoph (2003) NMDA receptors with reduced Ca2+ permeability and their ef- Funktion des cytoplasmatischen und nukleären ß-Catenins in fect on hippocampal long-term potentiation in mutant mice der Morphogenese von Hydra Astori, Simone (2006) Scholz, Corinna (2003) Modulation of GABAergic transmission in the cerebellar stel- Isolation und Funktion von mesodermalen Genen bei Nema- late cell network by neurotransmitter spillover and synaptic tostella. cross talk Kusserow, Arne (2005) Punnakkal, Pradeep (2007) Wnt genes from Nematostella Functional consequences of triheteromeric NMDA receptors in Guder, Corina (2005) hippocampal synapses Signalling molecules from Hydra Kuner, Rohini Bertulat, Bianca, (2008) Vodrazka Peter (2008) bHLH genes in Hydra Role of Plexin-B1/sema4D in hippocampal development Mättner, Robert (2008) Hirschberg, Alexandra (2008) Hydra transgenesis Role of Plexin-B/sema4D in corticogenesis Lengfeld, Tobias (2008) Hydra canonical wnt genes Kuner, Thomas Wimmer, Verena (2004) Kins Räumliche Verteilung und dynamische Organisation synapti- Kuan, Yung-Hui (2005) scher Vesikel in der Heldschen Calyx Characterization of APPBP2/PAT1a and Analysis of its interac- Abraham, Nixon (2006) tion with APP family Proteins Mechanisms of odor discrimination in the olfactory bulb of Szodorai, Anita (2006) mice Characterization of the transport machinery in axonal APP ve- Schleich, Wolfram (2006) sicles Clomeleon Biosensor-Mauslinien zur optophysiologischen Grübl, Tomas (2007) Dondzillo, Anna (2008) Analysis of the PAT1a binding motif in APP/APLPs and identifi- Active zone proteins Piccolo and Bassoon at the calyx of Held: cation of novel PAT1a binding partners affecting APP endocy- age-dependent localization and targeted in vivo perturbation tosis and processing Groh, Alexander (2008) Kirsch A giant driver synaptic connection in the cortico-thalamic pa- Becker, Michael (2008) thway of the rodent whisker system Der Elongationsfaktor (eEF1 A) und seine Beziehung zum Zyto- Kempf, Christian (2008) skelett in eukaryontischen Organismen 3D distribution of matrix proteins in the calyx of Held

185 Doctoral Theses

Mense Doganci, Beril (2006) Reinöhl, Jochen (2006) The generation and characterisation of mice with conditional ATP and capsaicin as mediators of muscle knock-out of the NMDAR subunit NR2B Dahlhaus, Anne (2007) Zivkovic, Aleksandar (2006) Synaptic input of rat spinal lamina I neuron The role of feedback and feed-forward perisomatic inhibition in the hippocampal gamma oscillations Monyer Vogt, Angelika (2007) Blatow, Maria (2003) Generation and analysis of transgenic mice expressing CRE re- A novel network of multipolar bursting interneurons in the combinase in defined interneurons neocortex Goengrich, Christina (2008) Filippov, Mikhail (2003) Protein-protein interactions of GAP junctions channels Tracing connexin26 expression using a genetic “knock-in” ap- Racz, Attila (2008) proach Establishing a relationship between molecular determinants of Barbe, Michael (2005) hippocampal oscillatory activities and cognitive behaviour The physiological characterization of Pannexin hemichannels Niehrs Coserea, Irina (2005) Excitotoxicity in the CNS in mouse mutants with manipulated Bilic, Josipa (2007) glutamate receptors Screening for nuclear reprogramming factors and analysis of DNA demethylation during in vitro myoblast differentiation Helbig, Ingo (2005) Labeling of electrical synapses in a transgenic animal model Hassler, Christine (2007) Characterization of developmental role and mechanistic func- Inta, Dragos (2005) tion of Kremen proteins Expression profile of 5-HT3-positive neurons in a transgenic mouse model Swaminathan, Suresh (2007) Signaling of the Wnt co-receptor LRP6 Tönges, Lars (2005) Generation and analysis of a trangenic mouse with fluores- Pollerberg cently labeled N-Methyl-D-Aspartate receptor 2D Peter, Jan-Christoph (2003) Arslan, Ayla (2006) Modulation der Aktivität des nikotinischen Acetylcholinrezep- Specifying molecular determinants of the subcellular targeting tors in transgenen Tiermodellen of synaptic and extrasynaptic GABAA receptors Henschel, Oliver (2004) Chen, Jing (2006) Funktion der muskelspezifischen Kinase MuSK: Analyse durch Cerebellar granule cell-specific deletion of the AMPA receptor konditionale Inaktivierung im transgenen Tiermodell subunit GluR-D gene Schmid, Christina (2004) Mihaljevic, André (2006) Identifizierung und Charakterisierung von AST-1 in C. elegans, Analysis of recombinant glutamate receptors in the hippocam- einem ETS Transkriptionsfaktor, der die axonale Wegfindung pus of the mouse reguliert Sandu, Cristina (2006) Kleber, Susanne (2005) The generation and the characterization of the TASK-3 knock- γ-Bestrahlung führt zu CD95-abhängiger Invasivität in apopto- out mice seresistenten Glioblastomzellen Timmermann, Patricia (2008) Sotiropulos, Ioannis (2006) Veränderung der N-methyl-D-aspartat-Rezeptoruntereinheit Identification of the cellular mechanisms underlying the con- 2A im Tiermodell tribution of stress and glucocorticoids to Alzheimer’s disease Bark, Christine (2006) pathology Connexin 36 vermittelt die synchrone Entladungstätigkeit von Neidhardt, Lukas (2007) Mitralzellen in den Glomeruli des olfaktorischen Bulbus Functional characterization of Lupli

186 Doctoral Theses

Uhlenhaut, Henriette (2007) ein Kandidatengen für die Transposition der großen Gefäße The role of the winged-helix transcription factor Fox12 des Herzens Knab, Christoph (2007) Häcker, Beate (2005) Axonal elongation on micro-patterned substrates Identifizierung von Zielgenen des Transkriptionsfaktors SHOX Avci, Hasan (2005) (Short Stature Homeobox Containing Gene) The role of cell adhesion molecules for axonal growth and ori- Endris, Volker (2004) entation Identifizierung und Charakterisierung von Genen für mentale Zelina, Pavol (2005) Retardierung/cognitive Funktion The role of cell adhesion molecule intracellular signalling for Sabherwal, Nitin (2006) axonal functions Functional characterization of missense and regulatory muta- Kinge, Parag (2005) tions in the human short stature homeobox gene (SHOX) Genetic analysis of proteins involved in the invertebrate ner- Schneider, Katja (2007) vous system development Funktionelle Charakterisierung des Homeoboxgens SHOX und Schwarz, Valentin (2005) Analyse eines transgenen Tiermodells Functional analysis of IgSF CAMs in C. elegans nervous system Spitzer, Jessica (2006) development Identifizierung und Charakterisierung von Interaktionspart- Sotiropulos, Ioannis (2006) nern des Transkriptionsfaktors SHOX2 Cellular and molecular role of stress for microtubules Rupp Maier, Bettina (2007) Klein, Holger The role of cell adhesion molecules DM-GRASP in the growth Mismatch negativity and musical aptitude in Alzheimer Di- cone sease Cavalcanti-Adam, Ada Elisabetha (2005) Kugel, Julia Spatial arrangement of ligands by nano-templates affects cell Neuromagnetic representation of comodulation masking re- adhesion and signalling lease Seiler, Markus (2007) Sönmez, Banu Program 3of5: Screening for complex patterns in potein seqen- Tracking the neuromagnetic responses in auditory cortex as ces the repetition rate of a sound is varied across the lower limit Schmitz, Caroline (2007) of pitch Genetic analysis of CAMs involved in axonal pathfinding Schuster Albrecht, Steffen (2008) Impact of nano-scale patterns of surface proteins on cellular Knirr, Matthias (2007) responses Analysis of the novel translational regulator Charybde at Dro- sophila NMJs Wolfram, Tobias (2008) Impact of nano-patterned IgSF-CAMs on neuronal adhesion Schütz and signalling Kirilov, Milen (2006) Rappold Characterization of HNF mutants in the mouse Frank, Bernd (2003) Ronzaud, Caroline (2007) Identifizierung, Charakterisierung und physikalische Kartie- Role of mineralocorticoid receptor in sodium retention in the rung Serotonin-Rezeptor-homologer Gene und Mutations- kidney analyse in Patienten mit neuropsychiatrischen Erkrankungen Elzer, Joachimq (2007) Muncke, Nadja (2003) Analysis of non-genomic action of steroid hormones Positionsklonierung und Charakterisierung von PROSIT240 -

187 Doctoral Theses

Erdmann, Gitta (2007) Pawlak, Verena (2004) Identification and characterization of mineralocorticoid recep- NMDA receptors with reduced Ca2+ permeability and their ef- tor target genes in the mouse hippocampus fect on hippocampal long-term potentiation in mutant mice Habermehl, Daniel (2008) Licznerski, Pawel (2004) Generation of knockdown mutations in signal-dependent Regulation of AMPA receptor function by RIL: a trafficking stu- transcription factors by RNA interference and lentiviral vectors dy based on the use of recombinant Sindbis and Lentiviral vec- Rieker, Claus (2008) tors in cultured neurons CREB/CREM function in dopaminergic neurons Freese, Simone (2004) Doxycyclin-kontrollierte Genexpression im ROSA26-Lokus der Schwaninger Maus. Zhang, Wen (2003) Kolleker, Alexander (2005) Einfluss von NF-κB auf die zerebrale Ischämie Trafficking regulations of AMPA receptor synaptic functions: Kleesiek, Jens (2005) GluR-Blong mediates juvenile synaptic plasticity & stargazin Die Wirkung des IKK-Inhibitors BMS-345541 auf die Genexpres- regulates the receptor desensitization sion von NF-κB-Zielgenen in der zerebralen Ischämie Hambsch, Boris (2005) Inta, Ioana (2005) Signaltransduktion der Intrazellulären Domäne der g-Proto- TWEAK and NF-κB in cerebral ischemia cadherine Loewer, Marc (2005) Dittgen, Gil Tanjew (2005) Takayasu-Arteriitis Optimization of Sindbis virus and Lentivirus expression sys- Herrmann, Oliver (2005) tems for genetic manipulation of neurons in vivo Regulation der Körpertemperatur und Neuroprotektion durch Zhu, Peixin (2006) endogenes Interleukin-6 während zerebraler Ischämie Different genetic ways for conditional gene regulation in the Schölzke, Marion (2005) mouse brain Die Aktivierung des Transkriptionsfaktors NF-κB in Körnerzel- Bonn, Stefan (2007) len nach exzitatorischer Stimulation mit Glutamat Processing and signalling of α- and γ-Protocadherins Keinert, Timo (2007) Marx, Verena (2007) Cannabinoid Rezeptor 2 in der zerebralen Ischämie, eine phar- Molecular alterations of AMPA receptors and their effects on makologische Studie am Mausmodell hippocampus dependent tasks Muhammad, Sajjad (2008) Bus, Thorsten (2008) High mobility group box 1, receptor for advanced glycation Switching excitatory neurotransmission in the adult hippo- end products and IkappaB kinase are potential pharmacologi- campus: a study with a conditional mouse mutant cal targets in cerebral ischemia Meyer Borgloh, Stefan (2008) Lang, Min-Fei (2008) GluR-A distribution and calcium signals in mouse neocortex A transgenic strategy to define SLCO1c1-expressing structures measured in vivo by 2-photon microscopy during brain development Cetin, Ali Haydar (2008) Seeburg Viral vectors for studying nervous system development and function Schleich, Wolfram (2006) Clomeleon Biosensor-Mauslinien zur optophysiologischen Be- Spanagel stimmung der intrazellulüren Chloridkonzentration in Nerven- Abarca, Carolina (2003) zellen The interaction of drugs of abuse with molecular components Shimshek, Derya Robert (2003) of the internal clock GluR-B: die wichtigste AMPA-Rezeptor-Untereinheit in der Kirschke, Christian (2003) Maus Studies on conditioned opioid withdrawal

188 Doctoral Theses

Perreau-Lenz, Stephanie (2004) Unsicker Control of the daily melatonin rhythm Adodo, Godfrey (2003) Molander, Anna (2005) Effects of Histone Deacetylase inhibitors on the expression of Role of glycine receptors in the regulation of dopamine activity AMPA receptor subunits and the alternative splice variants of and ethanol intake in the rat GluR2 Flip/Flop Vengeliene, Valentina (2005) Allmendinger, Alexandra (2004) Long-term voluntary alcohol consumption in the rat: interac- Structural and chemical alterations of adrenal chromaffin cells tion of genetic and environmental factors in mice lacking c-Ret Zieker, Derek (2006) Lenhard, Thorsten (2004) Gene expression profiling following exhaustive exercise Neuroprotektive und genregulatorische Funktionen von Glial Zghoul, Tarek (2007) Cell Line-Derived Neurotrophic Factor in einem in vitro Exzito- Altered circadian states and their retuned relationship with al- toxizitätsmodell des Hippokampus cohol Subramaniam, Srinivasa (2004) Zieker, Judith (2007) Regulators of cerebellar granule neuron degeneration Gene expression in patients suffering from posttraumatic Franke, Aylin (2005) stress disorder Role of BMP’s in triggering chromaffin cell development Spors Schmeer, Corina (2006) Expression of Growth Differentiation Factor-15 (GDF-15) in hu- Künsting, Thomas (2007) man gliomas and its functions in glioblastoma cell lines Testing an olfactory bulb model with dynamic input patterns Strzelzcyk, Adam (2007) Sprengel Charakterisierung der in vivo Funktion von growth/differentia- Shimshek, Derya Robert (2003) tion factor-15 für das Überleben adulter Motoneurone GluR-B: die wichtigste AMPA-Rezeptor-Untereinheit in der Carrillo-Garcia, Carmen (2008) Maus. Role of Growth/ Differentiation Factor (GDF) 15 in the regulati- Freese, Simone (2004) on of embryonic neural precursors Doxycyclin-kontrollierte Genexpression im ROSA26-Lokus der Krause, Sonja (2008) Maus Die Expression von Neurotrophinen und Neurotrophinzepto- Mihaljevic, André (2006) ren in Amygdala und Hippokampus von Mäusen Analysis of recombinant glutamate receptors in the hippocam- Esposito, Lorena (2005) pus of the mouse Sympathoadrenal progenitor heterogeneity Zhu, Peixin (2006) Gut, Philipp (2005) Different genetic ways for conditional gene regulation in the Analysis of adrenal-deficient mice mouse brain Oberle, Stephan (2006) Tucker LIFRß and cardiotrophin signalling for IML neuron development Sanno, Hitomi (2007) Postnatal control of cortical development by Rho GTPases Wick Maya Shaked (2008) Pfenning, Phillip-Niklas Role of histone deacetylases (HDACs) in embryonic neuroge- Functional characterization of hypoxia-induced invasiveness- nesis in the mouse forebrain related molecules Luger, Sebastian Glioma endothelium - stem cell interaction

189 Doctoral Theses

Wittbrodt Rembold, Martina (2007) Morphogenesis of the optic vesicle Ramialision, Mirana (2007) Identification and analysis of cis-regulatory modules in medaka eye development Souren, Marcel (2008) Six3 and gene regulatory networks in medaka eye development Witzemann Yampolsky, Pessah (2008) Regulation der Expression von AChR-Untereinheiten in transgenen Tiermodellen

190 Lectures, seminars and courses offered by IZN Investigators

Lectures, seminars and courses offered by IZN Investigators

Lectures

From evo-devo to comparative genomics; Evolution module of EMBL PhD course Detlev Arendt Cell biology III Hilmar Bading Cell biology IV Hilmar Bading Einführung in die Molekulare Psychiatrie Dusan Bartsch Psychosomatische Medizin und Psychotherapie Martin Bohus Medical physiology Andreas Draguhn Transportprozesse in biologischen Membranen Thomas Euler Einführungsvorlesung, Grundpraktikum C4 (Nerv/Sinne) Thomas Euler Clinical and cognitive neurosciences Herta Flor Medical psychology (MaReCuM) Herta Flor Sensory physiology – from ion channels to behaviour Stephan Frings Animal- and human physiology Stephan Frings Module Focus Bioscience 1 “Sensory physiology” Stephan Frings Module Focus Bioscience 2 “Physiology of the hippocampus” Stephan Frings Biology IV (Developmental Biology and Immunology) Thomas Holstein Molecular Developmental Biology Thomas Holstein Biology III (Molecular Cell Biology) Thomas Holstein Molekularbiologie I Stefan Kins Zellbiologie IV Stefan Kins Generation of Transgenic Mice and Joint Biotechniques Stefan Kins Microscopical Anatomy Joachim Kirsch Macroscopical Anatomy Joachim Kirsch Cell Biology Joachim Kirsch Integrated lecture (Biochem., Phys., Anat.) Thomas Kuner Molecular Biosciences Thomas Kuner Macroscopic anatomy, neuroanatomy, and histology Siegfried Mense Integrative Neuroanatomie/Neurophysiologie Hannah Monyer Neurovisionen: Synchrone Netzwerkaktivität und Plastizität im ZNS Hannah Monyer Medizinische Psychologie (Pschosoziale Grundlagen der Medizin) Hannah Monyer Pharmakologie und Toxikologie von Arzneistoffen Ulrike Müller Ringvorlesung Biologie A, Molekulare Biotechnologie Ulrike Müller

191 Lectures, seminars and courses offered by IZN Investigators

Molekulare Biotechnologie Ulrike Müller Begriffe der molek. Embryonalentwicklung der Wirbeltiere Christof Niehrs Methoden der molekularen Entwicklungsbiologie bei Wirbeltieren Christof Niehrs Biological Psychology Sabina Pauen Theories and Methods of Developmental Psychology Sabina Pauen Basics of Neuropsychology Sabina Pauen Development across the life-span Sabina Pauen Early childhood development: Applied Aspects Sabina Pauen Entwicklung des visuellen Systems Elisabeth Pollerberg Zelluläre und molekulare Neuroentwicklungsbiologie Elisabeth Pollerberg Human Genetics Gudrun Rappold Psychology and Neurophysiology of Hearing Andre Rupp EEG-MEG: Methods and Applications Andre Rupp Physiology Martin Schmelz Zyklusvorlesung Neurobiologie: Neuronale mikroRNAs Gerhard Schratt Lernen & Gedächtnis, Synapsen und Moleküle Christoph Schuster Nerv und Sinne (Begleitvorlesung zum tierphysiologischen Grundpraktikum GP-C4) Christoph Schuster Zellbiologie IV Ringvorlesung Christoph Schuster Pharmacology and Toxicology for students of pharmacy, biology, and chemistry Markus Schwaninger Pathophysiology Markus Schwaninger Molecular and cellular neurobiology Peter Seeburg Developmental Neurobiology Horst Simon Rezeptorklassen, Membrantransport, Exocytose, Endokrine Funktionen/Hormone Thomas Söllner Klinische Pharmakologie Rainer Spanagel Suchterkrankungen Rainer Spanagel Psychiatrie Rainer Spanagel Mouse Models in Neuroscience Research Rolf Sprengel Parkinson’s Disease: Causes and Therapy Kerry Tucker Neurobiologie Klaus Unsicker Neuroanatomie Klaus Unsicker Integrierte Vorlesung/Organe und Organsysteme) Klaus Unsicker Basics in neurooncology and neuroimmunology Wolfgang Wick Neurology Wolfgang Wick Numerische Algorithmen, Einführung in die Numerische Mathematik Gabriel Wittum Numerik 1: Diskretisierung partieller Differentialgleichungen Gabriel Wittum

192 Lectures, seminars and courses offered by IZN Investigators

Numerik 2: Schnelle Löser für große Gleichungssysteme Gabriel Wittum Wissenschaftliches Rechnen Gabriel Wittum

Seminars and Colloquia

Introduction to Biosciences Hilmar Bading Transformation, transfection, infection Hilmar Bading Instruments and techniques in Biosciences Hilmar Bading Neuronal plasticity Hilmar Bading Calcium signals in learning, survival, and death Hilmar Bading Modul Frontiers in Biosciences I: Modern methods in Neurobiology Hilmar Bading Doktorandenseminar Martin Bohus Studenten im Praktischen Jahr, Tutorium Martin Bohus Ausgewählte Kapitel der Psychosomatik Martin Bohus Neural Stem Cell Biology Francesca Ciccolini Instruments and techniques in Biosciences Francesca Ciccolini Medical physiology Andreas Draguhn Preclinical physiology Andreas Draguhn Visuelles System Thomas Euler Informationsverarbeitung in sensorischen Systemen Thomas Euler Gene, Gehirn & Kognition Christian Fiebach Methoden der kognitiven Neurowissenschaften Christian Fiebach Neuroanatomie des Belohnungssystems Christian Fiebach Kolloquium der Arbeitsgruppe Neurokognition und Kognitive Neurologie Christian Fiebach Seminar psychophysiology in clinical and health psychology Herta Flor Seminar neuropsychology Herta Flor Seminar psychology of pain Herta Flor Pain: Foundations, assessment, treatment Herta Flor Psychophysiological methods Herta Flor Anxiety disorders or addiction Herta Flor Medical psychology (MaReCuM) Herta Flor Sensory organs, sensory cells, sensory perfomance Stephan Frings Biochemistry and physiology of everyday life Stephan Frings Signal transduction in sensory cells Stephan Frings Module Focus Bioscience 1 “Sensory physiology” Stephan Frings

193 Lectures, seminars and courses offered by IZN Investigators

Module Focus Bioscience 2 “Physiology of the hippocampus” Stephan Frings Translational medicine in Psychiatry: from Animal Models to Therapy of Human Disorders Peter Gass Biochemical Baselines of Psychiatric Disorders Peter Gass Genomic model systems: evolution, function and disease Thomas Holstein Molecular mechanisms of evolution and development Thomas Holstein Progress in Molecular Developmental and Evolutionary Biology Thomas Holstein Advanced Biochemistry: Expression and study of signaling and adhesion proteins Thomas Holstein Genetik, Molekular-, Zell- und Neurobiologie der Alzheimerkrankheit Stefan Kins Aktuelle Probleme der Molekular- und Neurobiologie Stefan Kins Aktuelle Probleme der Genetik, Molekular-, Neuro- und Zellbiologie Stefan Kins Molekularbiologie und Genetik der Alzheimer Krankheit Stefan Kins Tiermodelle der Entwicklung, Regeneration und Degeneration in der Neurobiologie Stefan Kins Tiermodelle neurodegenerativer Erkrankungen Stefan Kins Molekularbiologie der Proteine begleitend zum Praktikum Molekularbiologie Stefan Kins Entwicklungsbiologie des Nervensystems Stefan Kins Neurodegeneration Stefan Kins Molecular biology, cell biology and genetics of Alzheimer`s disease Stefan Kins “Physiology” for Medical students (cardivascular, respiratory, urinary) Georg Koehr Graduate Students Seminar at the Max-Planck-Institute Georg Koehr “Physiology” for Medical students (Muscle and nervous system) Georg Koehr “Atherosklerose” for Medical students Georg Koehr Allgemeine Pharmakologie Rohini Kuner Physiology seminar Thomas Kuner Integrated seminar Thomas Kuner Introduction to biology students Thomas Kuner Molekulare Grundlagen neuropsychiatrischer Erkrankungen Hannah Monyer Tiermodelle der Entwicklung, Regeneration und Degeneration in der Neurobiologie Ulrike Müller Aktuelle Probleme der Neurobiologie und funktionellen Genomik Ulrike Müller Critical Reading in Developmental Psychology Sabina Pauen Research seminar for advanced students Sabina Pauen Infancy Research Sabina Pauen Bioethik Elisabeth Pollerberg Developmental neurobiology Elisabeth Pollerberg Entwicklung des Nervensystems von Modellorganismen Elisabeth Pollerberg How axons grow and navigate Elisabeth Pollerberg

194 Lectures, seminars and courses offered by IZN Investigators

Progress in developmental neurobiology Elisabeth Pollerberg Recent publications in developmental biology Elisabeth Pollerberg Einführung in das Studium der Biologie Elisabeth Pollerberg Einführung in die Entwicklungsneurobiologie Elisabeth Pollerberg Progress in Developmental Neurobiology Elisabeth Pollerberg Recent publications in Developmental Neurobiology Elisabeth Pollerberg Human Molecular Genetics Gudrun Rappold Human Genetics Gudrun Rappold Pharmacotherapy Markus Schwaninger Ionenkanäle – Strukturelemente und Funktion Peter Seeburg Excitatory neurotransmission Peter Seeburg Use of Software and Databases in Molecular Biology Horst Simon Disorders of the Motor System Horst Simon Developmental Neurobiology Horst Simon Proteine I und II, Verdauung/Lipide, Biochemie der Cancerogenese Thomas Söllner Addiction Biology Rainer Spanagel Biologie der Drogenabhängigkeit Rainer Spanagel 14th Jerusalem School in Life Sciences “The Sense of Smell” Hartwig Spors Vom Gen bis zur 3D Struktur des Genproduktes Rolf Sprengel Axon growth, synaptogenesis, plasticity Rolf Sprengel Neurogenesis in Central and Peripheral Nervous System Kerry Tucker The Growth Cone: Sensitivity and Guidance Kerry Tucker Epigenetics and Neurobiology Kerry Tucker Mikroskopische Anatomie Klaus Unsicker Neuroanatomie Klaus Unsicker Zellbiologie-Seminar Klaus Unsicker Modellierung des Transports durch Biogewebe Gabriel Wittum Modellierung der Signalverarbeitung in Neuronen Gabriel Wittum Informatik und Gesellschaft Gabriel Wittum Seminar for Graduate College students Veit Witzemann Graduate Students Seminar at the Max-Planck-Institute Veit Witzemann

195 Lectures, seminars and courses offered by IZN Investigators

Practical Courses

Evolution module of EMBL PhD course Detlev Arendt Experimental Physiology and Developmental Biology Hilmar Bading Introduction to gene transfer techniques Hilmar Bading Live imaging of mammalian cells Hilmar Bading Proteins Hilmar Bading Neuronal calcium signaling in plasticity and survival Hilmar Bading Electrophsiology of synaptic plasticity Hilmar Bading Modern methods in Neurobiology Hilmar Bading Modul Frontiers in Biosciences I: Modern methods in Neurobiology Hilmar Bading Psychosomatische Medizin und Psychotherapie Martin Bohus Neural Stem Cell Cultures Francesca Ciccolini Medical physiology Andreas Draguhn Neurophysiology Andreas Draguhn Cellular Neurophysiology for the “Studienstiftung des Deutschen Volkes” Andreas Draguhn Preclinical physiology Andreas Draguhn Biophysikalische Methoden, Elektro- und Optophysiologie/Retina Thomas Euler Tierphysiologie (Versuchsteil Nerv/Sinne) Thomas Euler Modern methods in Neurobiology Thomas Euler RNA and DNA Stephan Frings Human Physiology Stephan Frings Bioloab “Electrophysiology”, “Biochemistry of the senses“ Stephan Frings Animal physiology 1 Stephan Frings Animal physiology 2 Stephan Frings Experimental physiology Stephan Frings Module Focus Bioscience 1 “Sensory physiology” Stephan Frings Module Focus Bioscience 2 “Physiology of the hippocampus” Stephan Frings Experimental Embryology of Marine Invertebrates Thomas Holstein Signalling Molecules in Development Thomas Holstein Developmental Biology Thomas Holstein Experimental Developmental Biology Thomas Holstein Structure and Function of Signalling Molecules Thomas Holstein Advanced Biochemistry: Expression and study of signaling and adhesion proteins Thomas Holstein

196 Lectures, seminars and courses offered by IZN Investigators

Molekularbiologie (3 wö., ganztägig) Stefan Kins Molekularbiologie der Proteine (1 wö., ganztägig) Stefan Kins Methoden der Molekularbiologie und Genetik der Alzheimer Krankheit Stefan Kins Molecular biology, cell biology and genetics of Alzheimer`s disease Stefan Kins Laborpraktikum (6 wö., ganztägig) Stefan Kins Grundpraktikum C (Teil 3) Mikrobiologie Stefan Kins Generation of Transgenic Mice and Joint Biotechniques Stefan Kins Microscopical Anatomy Joachim Kirsch Macroscopical Anatomy Joachim Kirsch Cell Biology Joachim Kirsch “Physiology” for Medical students (cardivascular, respiratory, urinary) Georg Koehr “Physiology” for Medical students (Muscle and nervous system) Georg Koehr “Leitsymptome’’ for medical students Rohini Kuner “Spezielle Themen der Pharmakologie” for students of the biology faculty Rohini Kuner Macroscopic anatomy Thomas Kuner Microscopic anatomy Thomas Kuner Gross anatomy Siegfried Mense Histology Siegfried Mense Immunohistochemistry in the central nervous system Hannah Monyer In vitro and in vivo electrophysiological recording in the hippocampus Hannah Monyer Methoden der Zellkultur und Funktionellen Genomik Ulrike Müller Wirkstoffforschung Ulrike Müller Entwicklungsneurobiologie Elisabeth Pollerberg Transfektion des visuellen Systems des Huhnembryos in ovo Elisabeth Pollerberg Zelluläre und molekulare Neuroentwicklungsbiologie Elisabeth Pollerberg Untersuchungen zur Rolle des Zytoskeletts im Wachstumskegel Elisabeth Pollerberg Molecular Genetics Gudrun Rappold Human Genetics Gudrun Rappold Brain Cell Lysate and Immune Precipitation Gudrun Rappold Modul Frontiers in Biosciences I: Modern methods in Neurobiology Gerhard Schratt Frontiers in Biosciences Christoph Schuster Learning & Memory, Synapses and Molecules Christoph Schuster Physiological Analysis of Synaptic Plasticity Christoph Schuster Biochemistry of Synaptic Plasticity Christoph Schuster Learning & Memory in Drosophila Christoph Schuster

197 Lectures, seminars and courses offered by IZN Investigators

Tierphysiologisches Grundpraktikum Christoph Schuster Pharmacology Markus Schwaninger Pharmacology for biology students Markus Schwaninger Virus mediated gene transfer Peter Seeburg Histology Horst Simon Cell biology Horst Simon Addiction Biology Rainer Spanagel Verhaltens- und molekulare Untersuchungen zur Drogenwirkung Rainer Spanagel Membranphysiologie Hartwig Spors Physiologie Hartwig Spors Axon growth, synaptogenesis, plasticity Rolf Sprengel Gene transfer to the mouse brain, Purification of the Taq-Polymerase Rolf Sprengel Using EGFP to illuminate the mouse nervous system Kerry Tucker Molecular, cellular and organismic studies, Using EGFP to illuminate the mouse nervous system Kerry Tucker Neurology Wolfgang Wick Molecular and genetic tools for the analysis of medaka and zebrafish development (EMBO course) Jochen Wittbrodt Intensive course at the MPI for Medical Reseach Veit Witzemann Course for Graduate College Veit Witzemann

198 NWG Lehrerfortbildung

Lehrerfortbildungsveranstaltungen der ‚Plastizität des Nervensystems ‘ Neurowissenschaftlichen Gesellschaft - 17.03.2006

‚Neurowissenschaften in der gymnasialen Oberstufe’ 10.00 Prof. Andreas Draguhn, IZN, Institut für Physiologie: Im Rahmen der ‚Brain Awareness Week’ Funktionsprinzipien von Synapsen

‚Neues vom Gehirn’ 11:00 Prof. Hannah Monyer, Dr. Jakob von Engelhardt, IZN, Klin. Neurobiologie der Neurologischen Klinik: 15. 03. 2004 Neubildung von Nervenzellen - ein Mechanismus der Plastizität? 10.15 Hilmar Bading, IZN, Neurobiologie: Lernen 13:00 Prof. Hilmar Bading, IZN, Neurobiologie: 11.00 Oliver von Bohlen, IZN, Neuroanatomie: Das emotionale Kalzium: Schalter für Veränderungen im Gehirn Gehirn 14:00 PD Dr. Oliver von Bohlen und Halbach, IZN, Neuroanato- 13.30 Andrea Ludolph / Joachim Kirsch, IZN, Anatomie & Zell- mie: biologie: Zappelphilippsgehirn – das Aufmerksam- Morphologische Aspekte erfahrungsabhängigen Ler- keitsdefizitsyndrom nens

14.15 Klaus Unsicker, IZN, Neuroanatomie: Wachstumsfaktoren 15:00 Erfassung biologischer Messwerte von Probanden im für Hirnzellen Praktikum. Demonstration mit praktischen Übungen. Referent: Ferdi Oberheinrich, Firma ADInstruments, 15.00 Horst Simon, IZN, Neuroanatomie: Entwicklungsneuro- Spechbach b. Heidelberg biologie und Krankheit

15.45 Kerry L. Tucker, IZN, Neuroanatomie: Heilung von Läh- ‚Signalmoleküle des Nervensystems: neue Techniken und mungen – Regeneration von Nervenbahnen nach trau- Ergebnisse der molekularen und zellulären Neurobiologie ‘ matischen Verletzungen 23.03.2007

‚Neurobiologische Grundlagen kognitiver Leistungen‘ 10:00 Prof. Dr. Thomas Söllner, BZH/IZN, Univ. Heidelberg: Exo- und Endocytose an der neuronalen Synapse 04.03.2005 11:00 PD Dr. Mathias Klugmann, IZN, Neurobiologie: 10.00 Prof. Andreas Draguhn, IZN, Institut für Physiologie: Gentherapie für neurologische Erkrankungen Informationsverarbeitung in neuronalen Netzwerken 13:00 Prof. Dr. Gudrun Rappold, IZN, Humangenetik, Univ. 11:00 Dr. Rainer Friedrich, IZN, MPI für Medizinische Forschung: Heidelberg: Funktionsprinzipien des Riechsinnes Genetische Grundlagen der geistigen Retardierung

13:00 Dr. Andreas Frick, MPI für Med. Forschung: 14:00 Prof. Dr. Thomas Kuner, IZN, Medizinische Zellbiologie: Mechanismen der synaptischen Plastizität Gezielte Eingriffe in die Funktion von Synapsen des zentralen Nervensystems durch viralen Gentransfer 14:00 Dr. Oliver Dick, IZN, Institut für Neurobiologie: Das Auge: Einblicke ins Gehirn 15:00 Versuche zur Neuro- und Sinnesphysiologie im Unter- 15:00 Prof. Herta Flor, IZN, ZI Mannheim: Traumagedächtnis richt: Demonstration mit praktischen Übungen. Ferdi Oberheinrich, Firma ADInstruments, Spechbach b. Heidelberg

199 NWG Lehrerfortbildung

‚Entwicklung des Nervensystems‘ 14.03.2008

10:00 Prof. Dr. Jochen Wittbrodt, Zoologie, IZN, Univ. Heidel- berg: Evolution und Entwicklung des Auges

11:00 Prof. Dr. Thomas Holstein, Zoologie, IZN, Univ. Heidelberg: Entstehung des Nervensystems

13:00 Prof. Dr. Joachim Kirsch, Anatomie und Zellbiologie, IZN, Univ. Heidelberg: Entwicklung von zentralnervösen Syn- apsen

14:00 Prof. Dr. Kerry Tucker, Neuroanatomie, IZN, Univ. Heidel- berg: Entwicklung der Anatomie des Gehirns

15:00 Prof. Dr. Christoph Schuster, Neurobiologie, IZN, Uni Hei- delberg: Axonale Wegfindung und Plastizität im Nerven- system von Insekten

200 Seminars and Symposia

201 IZN Seminar Series

IZN Seminar Series Tanja Kögel Myosin Va-dependent transport and maturation of secretory The IZN hosts four seminar series. The bi-weekly ‘IZN Seminars’, granules that provide doctoral students or postdocs with an opportunity Jochen Kuhse to present their projects, alternate with the ‘Heidelberg Neuro- The regulation of cluster formation of NMDA-receptor NR1- biology Lectures’, in which renowned scientists from abroad talk splice variants in spinal neurons about their latest research. At the bi-weekly ‘GK 791 Seminars’ Volker Nimmrich and the monthly ‘SFB 488 Seminar: Progress in developmental High-frequency (~200 Hz) network oscillations in the rodent neurosciences’, IZN Investigators and their group members re- hippocampus: analysis of cellular and network mechanisms port on their current projects. in vitro In addition, the IZN organizes special lectures, international symposia and the annual IZN retreat at Kloster Schöntal/Jagst, which Heike Peterziel has become a tradition for all Heidelberg/Mannheim neuroscien- Molecular mechanisms of GDNF/TGF-ß cooperativity tists to meet and discuss science. Katharina Schindowski Growth factors in cerebral ischemia: Regulation and physio- IZN Seminars 2003 logical roles of GDF-15, FGF-2, and FGF-5 Andreas Schober Hilmar Bading Neurotrophic factors for preganglionic sympathetic neurons Nuclear calcium signaling in neuronal survival Christian Scholz Mascha Blatow Development of midbrain dopaminergic neurons in chicken A novel network of multipolar bursting interneurons generates theta frequency oscillations in neocortex Kerry Tucker A green mouse: Using GFP to study peripheral nervous sys- Karin Burau tem development c-ret signaling and neuronal diversification Peer Wulff Antonio Caputi Cell type-specific modulation of neuronal activity Anatomical and physiological characterization of hippocampal calretinin cells IZN Seminars 2004 Francesca Ciccolini Characterisation of late neural stem cells by direct isolation: Isabel Aller uncouplingof acquisition of EGF responsiveness and bias to- Selective ablation of the GABAA receptor g2 subunit from wards glia generation. cerebellar granule cells Andreas Draguhn Ayla Arslan The role of transmitter uptake and transmitter metabolism Synaptic and extrasynaptic GABAA receptors at central inhibitory synapses Peter Bengtson Jakob von Engelhardt Synchronised recurrent bursting of cultured hippocampal Generation of transgenic mice expressing EGFP in cholinace- neurons – a model for investigating long lasting plasticity. tyltransferase-positive neurons Karin Burau Elke Fuchs Development of cholinergic sympathetic neurons Conditional gene ablation in GABAergic interneurons Sabine Chourbaji Ulrich Hoheisel Behavioural and transgenic approaches in murine animal Consequences of changes in spinal and supraspinal cGMP models for depressive disorders for spinal neuronal activity. Francesca Ciccolini Katrin Huber EGF is a motogen for late development stem cells The development of the sympathoadrenal cell lineage Irina Coserea Role of NMDA receptor subunits in excitatory cell death

202 IZN Seminar Series

Suhua Deng Paola Sgado Functional analysis of plexin-B family members plexin-B1 The atypical homeoprotein Pbx1a controls late axonal path- and plexin-B2 finding of mesencephalic dopaminergic neurons Beril Doganci Frank Stief Conditional knock-out of the NMDA receptor subunit NR2B Altered inhibition-excitation-balance in hippocampal inter- Thomas Dresbach neurons in chronic temporal lobe epilepsy Targeting proteins to nascent synapses: Routes to branches Jens Strelau Anja Eder Characterization of growth-differentiation factor 15, a trans- Generating Ca2+ signals by photolysis forming growth factor beta superfamily member Gitta Erdmann Markus Uhrig Analysis of glucocorticoid signaling in the mouse brain by From development to neurodegeneration- Neural transdif- gene targeting ferentiation of human adult stem cells? & Gene expression profiling of human neuroblastoma cells exposed to A beta Nidhi Gakhar peptides Calcium Signalling in Neural Precursors Wei Zhang Daniel Gherbassi Muscarinic modulation of SK channel activity in nigra neu- Mining the human genome: Identification of genes with po- rons tential links to Parkinson’s disease Aleksandar Zivkovic Christine Hassler Characterization of excitatory synaptic transmission in GluR- Functional screen for interaction partners of Kremen protein D knock-out mice in Xenopus Embryos Ulrich Hoheisel IZN Seminars 2005 Inflammatory cytokines and neurotrophins as stimulants for group IV muscle afferents in the rat. Kambiz N. Alavian Katrin Huber Mechanism of cell death in engrailed deficient mesencepha- Role of Phox2B and MASH-1 in chromaffin cell development lic dopaminergic neurons Dragos Inta Oliver von Bohlen In vivo labeling of specific interneuronal populations Role of trkB in the maintenance of dendritic spines Bettina Maier Antonio Caputi The role of DM-GRASP in the developing visual system Analysis and characterization of two Calretinin-positive GA- Bergic cell populations in layer II/II of the mouse neocortex David McCormick Rapid activation of recurrent cortical networks: A basic fea- Francesca Ciccolini ture of cortical operation? Effect of CREB / CREM transcription factors in neural stem cell proliferation and self-renewal Corina Popovici GDF-15 - a TGF-ß superfamily member with possible implica- Geoffrey Drew tions in the pathology of malignant gliomas Substance P drives endocannabinoid signalling in midbrain periaqueductal grey neurons by enhancing glutamate re- Hitomi Sanno lease The GTPase Rho & nerve development Alexei Egorov Caroline Schmitz Muscarinic control of graded firing activity in medial tempo- Screening for genes controlling axon guidance in C. elegans ral lobe structures Andreas Schober Ulrike Engel, Harvard Medical School, Boston GDNF/TGF-ß synergy in a model of Parkinson’s disease Signaling to the cytoskeleton in neuronal outgrowth: The Christoph Schuster microtubule plus end binding protein Orbit/CLASP media- Experience-dependent synaptic plasticity in Drosophila tes axon guidance downstream of the tyrosine kinase Abl.

203 IZN Seminar Series

Uwe Ernsberger Peer Wulff Factors for cholinergic development of sympathetic neu- Functions of GABA-A receptors on interneurons rons Amanda Wyatt Kristin Hartmann / Christina Janista Retrograde NO-signalling activates presynaptic NMDARs at “Regulation of efficacy at GABAergic synapses by presynap- Drosophila NMJs tic GABA content” Ulrich Hoheisel IZN Seminars 2006 Tetrodotoxin (TTX) as a tool for studying the spinal connectivity of nociceptive afferents Isabel Aller The neurobiology of TASK-1 and –3 potassium channels Matthias Klugmann CNS gene therapy in a rat leukodystrophy model Martin Both Specificity of information transfer within the hippocampus Thomas Kremer Identifying novel active zone proteins Ali Cetin Monitoring activity dependent changes in somato-sensory Jochen Kuhse barrel cortex Activity-dependent shedding of the NMDA receptor glycine binding site by matrix metalloproteinase 3: A novel mecha- Jing Chen nism of postsynaptic plasticity Cerebllar granule cell-specific deletion of the AMPA receptor GluR-D gene Daniela Lambertz Influence of chronic myositis on spinal field potentials of TTX Oliver Dick resistant skin and muscle afferents in rat spinal cord NMDA receptor regulated shuttling of FOXOs in hippocampal neurons Heike Peterziel Cooperativity between members of the TGF-beta superfami- Geoff Drew liy in neuronal cells Regulation of synaptic inhibition by glutamate uptake in the midbrain periaqueductal grey Andrei Rozov LTP (Induction and termination) Olivia Dumitrescu Characterization of 5HT3a-expressing cortical interneurons Andrei Rozov in a transgenic mouse model Supression of inhibition by a direct action of cannabinoids on ionotropic GABAA receptors Marina Eliava Morphological study of gap junctions using Cx36-EGFP Maya Shaked transgenic mouse Neural differentiation of ES cells Uwe Ernsberger Horst Simon Neuronal differentiation and diversification – the sympathe- Progressive loss of nigral dopaminergic neurons in postnatal tic system and beyond engrailed mutant mice Nidhi Gakhar Jörn Steinert Modulation of neural precursor cell differentiation by neural Are quantal size variations strictly postsynaptic? activity Stefan Titz Alexander Groh Ammonium uptake and KCC2 function: Pathophysiological Targeted expression of fluorescent proteins in layer 5B neu- and developmental aspects rons reveals a strongly depressing corticothalamic giant syn- Simone Velte apse imparting low-pass filtering of cortical inputs Modulation of SK channel activity in nigral neurons by disin- Frank Hofmann hibition and internal chloride concentration Microelectrode arrays (MEAs) as a tool to study late phase Simon Wiegert plasticity ERK trafficking in hippocampal neurons

204 IZN Seminar Series

Ulrich Hoheisel IZN Seminars 2007 Intracellular recordings of rat dorsal horn neurons in vivo before, during and after intramuscular injections of nerve Kambiz Alavian growth factor The role of P75 in the death of engrailed deficient mesencephalic dopaminergic neurons Dragos Inta Distinct postnatal migratory streams of GAGAergic neurons: Florian Bähner regulation by serotonin via its ligand-gated ion channel Plasticity of hippocampal neurons in memory-related network oscillations Dragos Inta Postnatal pancortical migration of GABAergic neurons mo- Waleed Barakat dulated by serotonin 5-HT3 receptors RAGE in cerebral ischemia Jarek Jarosik Tiziana Cesetti FGF-2 has antidepressant properties in an animal model of Absence of functional GABA-A receptors in stem cells of the depression postnatal subventricular zone Carla Margulies Uwe Ernsberger Nuclear Calcium Signaling: a green light for long-term me- To build a neuron (cont.) mory in Drosophila Elke Fuchs Ulrich Misgeld Recruitment of parvalbumin-positive interneurons determi- The functional role of endocannbinoid CB1 receptors on nes hippocampal function and associated behaviour neostriatal medium spiny neurons Nadine Holter Heike Peterziel Functional maturation of developing interneurons in the F-spondin promotes neuron survival and differentiation molecular layer of mouse dentate gyrus Alexei Ponomarenko David Lau Recruitment of interneurons shapes synchronous states of Analysis of transcription network involved in neuronal survival the intact hippocampus Volker Mack Hitomi Sanno A novel synaptic protein alters AMPA receptor-mediated Inducible expression of Rho GTPase modulators in the deve- transmission loping murine nervous system Daniela Mauceri Christian Scholz SAP97: Physio-pathological functions of a cargo protein K-ATP channels as a pharmacological target to rescue midbrain Andrei Rozov dopaminergic neurons from toxic insult nervous system Two Calretinin-positive GABAergic cell population in layer 2/3 Gerhard Schratt of mouse neocortex provide different forms of disinhibition MicroRNAs in synapse development Andreas Schober Stefanie Schuhmacher Zuckerkandl´s organ - re-visited RNA interference in neural stem cells Yongjoon Suh Jörg Steinert Effect of Dlx2 on neural precursor cells from subventricular Role of postsynaptic glutamate receptors in the experience- zone and hippocampus dependent activation of presynaptic NMDA Kerry Tucker Malte Wittmann Histone acetylation controls embryonic forebrain neuroge- Activity-dependent control of the geometry of neuronal nuclei nesis Wolfgang Wick Molecular basis of a cellular therapy for glioblastoma Nina Wittenmeyer Neuroligin 1 promotes maturation of the cytomatrix of active zones via postsynaptic interactions

205 IZN Seminar Series

IZN Seminars 2008 Heidelberg Neurobiology Lectures Nixon Abraham Synaptic inhibition accelerates odor discrimination in mice 2003 Julieta Alfonso Silvia Arber, Basel Neurogenesis and widespread forebrain migration of dis- Molecular pathways controlling sensory-motor connectivity tinct GABAergic neurons from the early postnatal subventri- in the spinal cord cular zone Elena Cattaneo, Milan Raphael Blüm Huntingtin function in Huntington’s Disease Glycine receptor activity affects intracellular signaling pa- Georg Dechant, Innsbruck thways in cultured neurons Neurotrophic factors and sympathetic neurotransmitter Isabel Brachmann plasticity Imaging of spinal nerve outgrowth in normal and semapho- Helmut Haas, Düsseldorf rin 3A mutant mouse embryos Histamine in the brain Carmen Carrillo Uwe Heinemann, Berlin Growth/differentiation factor (GDF) 15 regulates cell cycle Excitotoxical injury in nerve cells: involvement of mitochondria exit of secondary progenitors in the developing mouse ganglionic eminence Etienne Hirsch, Paris Glial cells and apoptosis in Parkinsonian syndromes Roberto Fiore Function of a large microRNA cluster in activity-dependent Kai Kaila, Helsinki neuronal development GABA: an exciting inhibitory neurotransmitter Florian Freudenberg Philipp Kahle, München The role of hippocampal AMPA receptors in an animal model Pathological features of a-Synuclein in Parkinson’s Disease of depression and related disorders Lydia Haussmann Hans Lassmann, Wien Protrusion and Retraction - Implications of the interaction Basic mechanisms of brain inflammation: their relevance for between MEGAP and the Ena/VASP binding protein Lamelli- the pathogenesis of multiple sclerosis and other human in- podin in lamellipodial dynamics flammatory CNS diseases. Alexander Lehmann Claudia Lodovichi, Durham Effects of a defined oligomeric Aß-species on synaptic activity New insights into olfactory system organization Sasidhar Murikinati Markus Missler, Göttingen CB2 receptor activation in cerebral ischemia alpha-Neurexins function as organizer molecules that couple Ca2+ -channels to synaptic transmission Pier Giorgio Pacifici Characterization of a mouse model lacking AChR activity du- Richard Olsen, Los Angeles ring embryonic development Structure, Function, and Plasticity of GABA-A Receptors Jan-Marek Weislogel Ole Paulsen, Oxford Visualisation of spatial distinct calcium signals in Hydra Network oscillations and synaptic plasticity in the hippocampus Sascha Weyer In vivo analysis of APP functional domains Frank Pfrieger, Strasbourg Cholesterol homeostasis and function in CNS neurons Sabrina Zechel Putative factors for the development and maintenance of Olaf Pongs, Hamburg the nigrostriatal dopaminergic system M-channels in the central nervous system - determinants of neuronal excitability

206 IZN Seminar Series

Jochen Roeper, Marburg Volkmar Lessmann, Mainz To spike or to burst - ionic mechanisms in control of electri- Intracellular targeting and synaptic secretion of neurotro- cal activity in dopaminergic neurons phins Dietmar Schmitz, Berlin Siegrid Löwel, Magdeburg Comparison of different forms of synaptic plasticity in the Experience-dependent plasticity of intracortical circuitry hippocampus and functional maps in the visual cortex Jörg Schulz, Tübingen Paolo Malatesta, Parma Molecular pathways of neuronal death: From mechanisms to Glial cells generate neurons: cellular and molecular mecha- treatment nisms of neurogenesis Marten Smit, Utrecht David McCormick, Boston Molecular mechanisms of midbrain dopamine neuron deve- Rapid activation of recurrent cortical networks: a basic fea- lopment ture of cortical operation? Ueli Suter, Zürich Thomas Misgeld, St. Louis Neuron-glia interactions: Lessons from development and di- Imaging the dynamics of axon removal in development and sease of the nervous system disease Roger Traub, Brooklyn Hans-Werner Müller, Düsseldorf Contrasting effects, on the generation of gamma oscillations, Long-distance axon regeneration and functional improve- of electrical coupling between principal cells and between ment in spinal cord injury interneurons Harald Neumann, Göttingen Eberhard Weihe, Marburg Molecular mechanism of inflammatory axonal injury Chemical coding of neurotransmission: The vesicular trans- Juha Partanen, Helsinki porter connection Intercellular signalling at the boundary between developing mid- and hindbrain Heidelberg Neurobiology Lectures 2004 Thomas Perlmann, Stockholm Eero Castren, Helsinki Nuclear receptor signaling in dopamine neuron develop- Physiological, pathophysiological and pharmacological ef- ment and survival fects of neurotrophins in adult brain Michael Sendtner, Würzburg Richard Dyck, Calgary Role of B-raf and BAG-1 in neurotrophin signalling Zinc as a neurotransmitter - Implications for plasticity and Esther Shohami, Jerusalem pathology Targeting intracellular mechanisms: a new approach to treat- David I. Graham, Glasgow ment of traumatic brain injury Traumatic brain injury - What is new? Christine Stichel-Gunkel, Bochum Mike Gutnick, Jerusalem M. Parkinson: on the way to a new therapy Thalamocortical axons release serotonin during cortical cir- Roger Traub, New York cuit development Model of a thalamocortical column, exhibiting spindles, gam- Reinhard Jahn, Göttingen ma oscillations, and a variety of seizure phenomenologies Molecular mechanisms of neuronal exocytosis Antoine Triller, Paris Avihu Klar, Jerusalem Diffusion dynamics of the postsynaptic glycine receptor at a Transcriptional and post translational control of axon single molecule level guidance Klaus Willecke, Bonn Martin Korte, Martinsried Expression and functions of connexins in mouse brain, inclu- Neurotrophins and activity dependent synaptic plasticity ding retina

207 IZN Seminar Series

Gabriel Wittum, Heidelberg Yves DeKoninck, Québec Reconstruction of neuron geometrics from 2-photon micro- Altered chloride homeostasis as a substrate of chronic pain scopy data Michael Frotscher, Freiburg Laminating the Hippocampus Heidelberg Neurobiology Lectures 2005 Toshitaka Fujisawa, Mishima Nail Burnashev, Amsterdam Peptidomic approach to identify novel neuropeptides Inhibitory amino acid receptors in the CNS neurons as tar- Heiko Luhmann, Mainz gets for direct action of cannabinoids Oscillatory network activity in immature neocortical net- Johan Ericson, Stockholm works Application of developmental determinates to produce au- Yoshiaki Kidokoro, Maebashi thentic midbrain dopamine neurons from stem cells Multiple types of Ca2+ channels involved in exocytosis and Bernd Fakler, Freiburg endocytosis at the Drosophila neuromuscular junction Charakterisierung und physiologische Bedeutung der Mulit- Thomas Klausberger, Oxford proteinkomplexe von Kv1- und SK2-Typ Kaliumkanälen Network oscillations and GABAergic interneurons in the hip- Andreas Faissner, Bochum pocampus Structure and functions of the microenvironment in neural Claudio Rivera, Helsinki stem cell development and neuron-glia interactions There is more to KCC2 than chloride regulation Kurt Gottmann, Düsseldorf Björn Scheffler, Gainesville Role of the synaptic adhesion molecule N-cadherin in presy- Neural stem cells: “The good, the bad, and the ugly” naptic organization and function Philippe Vernier, Gif-sur-Yvette Christian Hübner, Hamburg Organization and differentiation of dopamine systems in Lessons from cation-chloride cotransporter knockout mice protochordates and vertebrates : An evolutionary view Dimitri Kullmann, London Manuela Zaccolo, Padova Synaptic plasticity in hippocampal interneurons: mecha- Real-time imaging of cAMP: visualization of the spatial and nisms and roles in circuit computations temporal dynamics of intracellular signalling Andreas Lüthi, Basel Input-specific mechanisms of synaptic plasticity in the late- Heidelberg Neurobiology Lectures 2007 ral amygdala Marina Pizzi, Brescia George Augustine, Durham NF-kappaB factors in the control of neuronal cell death Synaptic biophotonics: using optogenetic approaches to study brain circuitry Alain Prochiantz, Paris Engrailed and Otx2 transcription factors as signaling mole- Clive Bramham, Bergen cules in axon guidance and synaptic plasticity BDNF and control of LTP consolidation in the intact brain Gudrun Rappold, Heidelberg Emilio Carbone, Torino MEGAP, mental retardation and the cellular basis of cognition Calcium channels in chromaffin cells: new role for T- and L- types Gal Richter-Levin, Haifa Stress and Amygdala modulation of memory-related proces- Giorgio Carmignoto, Padova ses in the hippocampus Astrocyte-neuron dialogue promotes neuronal synchrony Thomas Deller, Frankfurt Heidelberg Neurobiology Lectures 2006 Reshaping the nervous system - dendritic reorganization of Marlene Bartos, Freiburg dentate granule cells following denervation Shunting inhibition improves robustness of gamma oscillati- Thomas Euler, Heidelberg ons in hippocampal interneuron networks by homogenizing Dendritic processing in the retina firing rates

208 IZN Seminar Series

Christian Haass, München David L. Van Vactor, Boston The molecular clockwork of Alzheimer’s Disease Signaling mechanisms that regulate synapse development Helmut Kettenmann, Berlin in Drosophila News from the glial world Martin Korte, Braunschweig The Yin and Yang of Neurotrophin receptor signalling in the SFB 488 Seminars: process of synaptic plasticity: results and fairy tails Progress in developmental neurosciences Georg Kuhn, Göteborg Neurogenesis in the adult brain under pathological conditions SFB 488 Seminars 2003 Martin Schmelz, Mannheim Hilmar Bading C-nociceptors in chronic pain patients - translational studies MAP kinase signaling in neuronal network plasticity Dirk Feldmeyer Heidelberg Neurobiology Lectures 2008 Structural and functional changes in the developing barrel Laszlo Acsady, Budapest cortex A fresh view on the thalamus: excitatory and inhibitory con- Rainer Friedrich trol of higher order thalamic relays Function and development of olfaction and mechanotrans- Yves Barde, Basel duction in zebrafish Using embryonic stem cells to study neural development Sheriar Hormuzdi Dan Ehninger, Los Angeles Neuronal intercellular channel-forming proteins : Molecular Mouse models of tuberous sclerosis: cognitive and behavio- investigations, functional implications ral aspects Katrin Huber Charles ffrench-Constant, Cambridge The development of the sympathoadrenal cell lineage Regulation of neural stem and precursor behaviour during Rohini Kuner development and repair by extracellular matrix Expression and functional implications of plexin-family Christoph Kellendonk, New York members in the developing nervous system Modeling schizophrenia endophenotypes in mice Bingyu Mao Andreas Püschel, Münster Kremen2 modulates Dickkopf2 during Wnt/LRP6 signaling How to make a neuron: signaling pathways directing axon Rosanna Parlato formation Generation of CREB mutations in catecholaminergic neurons André Rupp, Heidelberg Christoph Peter Pitch-Perception: Psychoacoustics, modelling, and neuroma- Generation of a transgenic mouse model for the slow chan- gnetic representation in the auditory cortex nel congentital myasthenic syndrome Paul Saftig, Kiel Daniel J. Spergel Proteolytic systems and their functions in neurobiology: Lyso- Glutamate receptors and the attainment of fertility somes, autophagy and proteolysis at the plasmamembrane Stefan Titz Andreas Schäfer, Heidelberg Does GABA promote the developmental switch of its own Mechanisms of odor discrimination in mice response? Vincent Torre, Triest Bill Wisden Force generation in neurons: elementary events and compu- Building GABAergic synapses tational properties Jochen Wittbrodt Control of proliferation and differentiation during early vertebrate eye development

209 IZN Seminar Series

SFB 488 Seminars 2004 Rainer Friedrich Optical measurements of spatio-temporal activity patterns Oliver Henschel in the brain MuSK is required to maintain postsynaptic organisation at Oliver Henschel the neuromuscular junction Synapse degradation and formation of new synapses upon Thomas Holstein conditional inactivation of MuSK Genetic regulation of neurogenesis in cnidarians Katrin Huber Emil Karaulanov Role of the adrenal cortex in chromaffin cell development Transcriptional regulation of BMP4 synexpression in Xeno- Martin Kluska pus in vitro, in vivo and in silico analyses Functional relevance of the GDP-GTP exchange factor colly- Rohini Kuner bistin interaction with the Map4-Kinase4 on the generation Sema4D-Plexin-B signaling in the development of hippo- and morphology of inhibitory synapses campal and cortical neurons Rohini Kuner Jun Li Semaphorin4D-plexin-B1 modulate neuritogenesis and Early functional development of chemotopy in the zebrafish neuronal migration in the developing forebrain olfactory system Ulrich Misgeld Oliver Schlicker Development of inhibition in substantia nigra GABAA-receptors in dendritic membrane traffic Alexandra Moers Stefan Titz G12/G13 are critically involved in cerebellar development in Functional consequences of KCC2 overexpression in cultu- mice red hippocampal neurons G. Elisabeth Pollerberg Kerry Tucker MAP1B phosphorylation in navigating growth cones Manipulating axonal outgrowth in mouse nervous system Derya Shimshek development GnRH neuron specific gene manipulations Tillmann Weber Kerry Tucker Transgenic mouse models of serotonergic deficits in depression Reverse & forward genetic analysis of peripheral nerve deve- Nina Wettschureck lopment Gq/G11-mediated signalling is required for endocannabino- Robert Waltereit id-mediated neuroprotection Altered episodic-like memory and impaired synaptic plasti- Tim Wintermantel city in a rat model of Tuberous Sclerosis Genetic dissection of estrogen receptor function in the ner- Bill Wisden vous system K2P channels and neuronal cell excitability William Wisden Jochen Wittbrodt Targeting GABA-A receptors to synapses Triggering neurogenesis in the retina Jochen Wittbrodt The role of Rx3 in vertebrate optic vesicle evagination SFB 488 Seminars 2006 Sheng-Jia Zhang Transcriptional profiling of activity-dependent gene expres- Peter Bengtson sion in cultured hippocampal neurons Somatic and dendritic calcium signaling during LTP induction Gary Davidson SFB 488 Seminars 2005 New screen, new gene: identification of an LRP6/Arrow kinase Volker Endris Dirk Feldmeyer Functional characterisation of the mental retardation associ- Neuronal connectivity in the neocortex of reeler mice ated GAP protein, MEGAP

210 IZN Seminar Series

David Engblom Stefan Offermanns Analysis of drug addiction using mutations specific to cells G_12 /G_13 -mediated signaling is required for the develop- of the dopamine system ment of cerebral and cerebellar cortices Corina Guder G. Elisabeth Pollerberg An ancient Wnt-Dickkopf-antagonism in Hydra Role of microtubule-associated protein APC for axonal steering Katrin Huber Christoph Schuster The development of the sympathoadrenal lineage A Myosin-driven postsynaptic signaling complex? Rohini Kuner Marc Willaredt The role of Plexin-B1 and Semaphorin 4D in branching mor- A role for ciliary function in cortical development phogenesis Jochen Wittbrodt Juan-Ramon Martinez-Morales The early interaction of Atrophin-2 and Six3 during forebrain The medaka mutation ojoplano disrupts a novel vertebrate patterning determines the late pathfinding of retinal axons gene with a fundamental role in tissue morphogenesis Pessah Yampolsky Alexandra Moehrs AChR channel conversion and AChR-adjusted neuronal sur- G-protein mediated signalling in cortical development vival during embryonic development Karsten Thelen Intracellular impact of cell adhesion molecule DM-GRASP on SFB 488 Seminars 2008 axon elongation and navigation Sidney Cambridge Kerry Tucker Doxycycline-dependent, photoactivated gene expression at Control of cortico-striatal neurogenesis by histone deacetyl- cellular resolution in eukaryotic systems transferases Volker Endris The inhibitory effects of MEGAP on actin dynamics SFB 488 Seminars 2007 Nadine Holter Hilmar Bading Inhibition in the developing dentate gyrus A nuclear calcium-regulated genomic survival program Milen Kirilov Volker Endris Neuroendocrine regulation of reproduction: Role of the est- MEGAP constitutes a microtubule associated protein widely radiol receptor and GPR54 expressed in the developing nervous system Ulrich Misgeld Roberto Fiore Synaptic and non-synaptic inhibition in juvenile GABA neu- Function of a large microRNA cluster in activity-dependent rons of substantia nigra neuronal development Marcel Souren Kristin Hartmann Identification of upstream factors for the differentiation of Plasticity of GABAergic synapses retinal ganglion cells Katrin Huber-Wittmer Konstantin Khodosevich Development of the sympathoadrenal lineage Major signaling pathways in migrating neuroblasts Rohini Kuner Karsten Thelen Role of neuronal plexin-B proteins in migration and pattern Role of the 3’UTR of DM-GRASP for local protein translation formation in vivo Klaus Unsicker Hai-Kun Liu Development and degeneration of extra-adrenal chromaffin The nuclear receptor tailless (tlx) is expressed in adult neural cells: Mode of cell death stem cells and required for their generation Thomas Worzfeld Analysis of plexin-B2 signalling in vivo

211 IZN Seminar Series

Graduate College 791 Markus Schwaninger Acute neurodegeneration in cerebral ischemia “Neural Development and Degenerative Peter Seeburg Processes: Basic Research and Clinical Glutamate receptors in the developing brain Implications” Klaus Unsicker TGF-ßs: Multifunctional growth factors with important roles in neural development and maintenance 2003 Klaus Unsicker Development of the nervous system and related malforma- Dusan Bartsch tions Learning, memory and gene expression William Wisden Francesca Ciccolini Is neurotransmission important for the developing brain? Stem cells of the central nervous system Veit Witzemann Thomas Euler / Group Winfried Denk Neurological diseases at the neuromuscular junction Signal processing in the mammalian retina Veit Witzemann Marc Fatar / Group Michael Hennerici Formation of synapses at the neuromuscular junction- Ultrasound treatment in animal stroke models a change of the neurocentric view Dirk Feldmeyer Neuronal networks in the neocortex – structural and functio- Graduate College 791 2004 nal properties Rainer Friedrich Dusan Bartsch Synchronization of neuronal ensembles: mechanisms and Learning to forget: memory extinction functions Guest Speaker: Ralf Baumeister, Freiburg Tobias Hartmann C. elegans models for the functional analysis of human ge- Alzheimer’s disease nes involved in neurodegenerative diseases Fritz A. Henn Stefan Berger / Group Günther Schütz Neurogenesis in affective disorders Analysis of mineralocorticoid and glucocorticoid receptor function in brain by gene targeting Harald Hutter Axon guidance signals: Molecules and mechanisms Francesca Ciccolini Development of mammalian interneurons Thomas Lemberger / Günther Schütz Group Genetic dissection of the role of the CREB transcription fac- Dirk Feldmeyer tor in neuronal survival Methods to study neurotransmission Ulrich Misgeld Rainer Friedrich The developmental change of the GABA response from de- Topographic maps in the nervous system to hyperpolarizing Tobias Hartmann Christof Niehrs Cellbiological basis for Alzheimer’s disease Mechanisms of antero-posterior axis formation Harald Hutter Stefan Offermanns Transcriptional networks controlling neuronal differentiati- G-protein mediated signaling in the developing and adult on nervous system Guest Speaker: Dietmar Kuhl/Molecular Neurobiology, Berlin Andrei Rozov Learning about activity dependent genes Pre- and postsynaptic mechanism of facilitation Bernd Kuhn /Group Winfried Denk / MPI Voltage-sensitive dyes: Basic principles and applications

212 IZN Seminar Series

Ulrich Misgeld Tobias Hartmann Monitoring KCC2 function during the developmental switch Neurons, lipids and neurodegeneration of the GABA response Fritz A. Henn Hannah Monyer The pathophysiology of depression Control of development by RNA processing Fritz A. Henn Guest Speaker: Zoltan Nusser, Budapest Use and misuse of animal models of brain diseases Short-term plasticity of excitatory and inhibitory synapses Harald Hutter Stefan Offermanns The importance of pioneer axons for neuronal circuit formation Functions and mechanisms of plexin-mediated signalling in Rohini Kuner the nervous system Basic mechanisms in physiological and pathological pain G. Elisabeth Pollerberg Ulrich Misgeld Studies in the avian embryo visual system Can we still learn from (substantia nigra) slices? Andrei Rozov Christof Niehrs Voltage and ligand gated ion channels Regulation of antero-posterior neural patterning during ear- Markus Schwaninger ly Xenopus development Mechanisms of neuronal apoptosis G. Elisabeth Pollerberg Peter Seeburg he tip of the axon: Molecules on and in the growth cone Long term potentiation Gudrun Rappold Guest Speaker: Traub Roger, New York Genetic etiology of mental retardation Evidence for electrical couplingbetween the axons of pyra- Andrei Rozov midal neurons, and the role of such coupling in generating Metabotropic receptors and synaptic plasticity oscillations Günther Schütz William Wisden Analysis of nuclear receptor function by gene targeting New techniques to regulate neuronal activity Markus Schwaninger William Wisden Clinical problems in basic stroke research Homeostasis in the developing brain Klaus Unsicker Veit Witzemann The TGF-ß superfamily of growth factors and their roles in Ion channel diseases: from men tmice the nervous system William Wisden Graduate College 791 2005 Silencing neurons in development Dusan Bartsch Veit Witzemann Epigenetic regulation of gene expression in the brain The value of toxins in science and medicine Francesca Ciccolini Clinical implication of neural stem cell research Graduate College 791 2006 Andreas Draguhn Hilmar Bading High-frequency network oscillations: cellular neuroscience Calcium signaling in the cell nucleus with some cognitive “meaning”? Dusan Bartsch Guest Speaker: Wolfgang Driever, Freiburg Animal models in psychiatric research Developmental neurogenetics in zebrafish: Cell specification in the dopaminergic and noradrenergic systems Dusan Bartsch Epigenetic regulation of gene expression in the brain Rainer Friedrich Calcium imaging in the brain

213 IZN Seminar Series

Francesca Ciccolini Graduate College 791 2007 Effect of neurotransmitter on neural precursor differentiation and proliferation Hilmar Bading Nuclear calcium signaling and activity-regulated adaptive Gary Davidson / Christof Niehrs group responses in the nervous system Mechanisms and regulation of cell movement Dusan Bartsch Andreas Draguhn Memory extinction Plasticity of GABAergic synapses - role of transmitter concentration Andreas Draguhn Analysis of neuronal networks Rainer Friedrich Genetic tools for probing and manipulating the function of Andreas Draguhn neuronal circuits Mechanisms and meaning of high-frequency network oscillations Peter Gass Genetically induced mouse models of stress-sensitivity and Guest Speaker: Julietta Uta Frey stress-resistance Cellular memory consolidation: associative requirements Ulrich Misgeld Peter Gass Electrophysiological studies on substantia nigra neurons in Neurobiological concepts of depressive disorders slices Guest Speaker: Martin Korte Hannah Monyer Learning and memory: What’s LTP got to do with it? GABAergic interneurone diversity in the CNS Guest Speaker: Martin Korte Ulrike Müller The Ying and Yang of Neurotrophin receptor signalling in the Functions of the Amyloid precursor protein gene family process of synaptic plasticity: results and fairy tails Stefan Offermanns Ulrich Misgeld G-protein mediated signalling in the nervous system SK channels control the excitability of juvenile GABA output neurons in substantia nigra slices G. Elisabeth Pollerberg The secret (intracellular) life of cell adhesion molecules: A Ulrich Misgeld glance behind the curtain Synaptic inhibition in substantia nigra Gudrun Rappold Hannah Monyer Mental retardation Development of GABAergic interneurones Andrei Rozov Guest Speaker: Klaus Nave Release probability versus response probability - two dimen- Molecular mechanism of axon-glia interactions and myelin sions of short-term synaptic plasticity disease Günther Schütz Guest Speaker: Klaus Nave Genetic analysis of steroid hormone receptor function Mouse models of neurological diseases Peter Seeburg Christof Niehrs Excitatory neurotransmission Early embryonic neural patterning in Xenopus Markus Schwaninger Stefan Offermanns Signaling through the transcription factor NF-kappaB Mechanisms and roles of semaphorin/plexin signaling in the nervous system Klaus Unsicker Transforming growth factor-beta: multifunctional and syner- G. Elisabeth Pollerberg gistic signaling in the nervous system Inside axons: The role of the cytoskeleton Veit Witzemann Gudrun Rappold Signals from nerve and muscle that regulate the develop- New insights into neurological disorders ment of neuromuscular junctions

214 IZN Seminar Series

Günther Schütz Gudrun Rappold The role of the nuclear receptor tailless in neurogenesis and New insights into mental retardation brain tumor formation Günther Schütz Markus Schwaninger Estrogen receptor function in neuroendocrine regulation of Anoxic tolerance in turtles and other vertebrates. What can reproduction we learn for neurological diseases? Markus Schwaninger Markus Schwaninger NF-kB signaling in health and disease Mechanisms of brain damage and repair in cerebral ischemia Peter Seeburg Peter Seeburg Molecular determinants of fast synaptic neurotransmission Excitatory neurotransmission Klaus Unsicker Klaus Unsicker GDF-15, a novel TGFß with functions in postnatal neuron Specification of neuronal and endocrine derivatives of the survival and myelination neural crest Klaus Unsicker Veit Witzemann Recent progress in understanding neural functions of GDF-15 ynaptic plasticity at neuromuscular junctions Veit Witzemann Receptor subunit switches are universal components of syn- Graduate College 791 2008 apse development Hilmar Bading Nuclear calcium signaling in learning and survival Seminars of the Dusan Bartsch DFG ’Forschergruppe’ FOR 302/2-1 Voltage gated calcium channels in normal and pathologic brain function 2003 - 2006 Francesca Ciccolini Andrzej Pilc, Beata Legutko, Krakow Characterization of SVZ neural stem cells Glutamate and depression Andreas Draguhn Dr. Madalina Stanciu, Pittsburgh Different recording methods and typical data in cellular elec- Mitogen-activated protein-kinases (MAPK) signaling in neu- trophysiology ronal death Jakob von Engelhardt / Hannah Monyer group Synaptic plasticity Neurobiology Seminars Peter Gass Translational models in psychiatry 2003-2005 Guest Speaker: Helmut Kettenmann, Berlin “Concepts in neuroscience” lessons from history Nikolaus J. Sucher, Hongkong University of Science and Technology Guest Speaker: Edvard Moser, Trondheim, Norway The NMDA receptor complex: A target for traditional chinese Grid cells and spatial representation in entorhinalhippocam- medicine pal neural networks Valentin Stein, UC San Francisco Ulrike Müller Cellular and molecular aspects of synaptic plasticity: PSD-95 Mouse models of neurodegenerative diseases mimics LTP Christof Niehrs Dmitri Tkachev, The Babraham Institute, Cambridge, U.K. Molecular mechanisms of early antero-posterior patterning Expression profiling in Schizophrenia: An integrative com- G. Elisabeth Pollerberg plementary approach Signalling inside and outside of growth cones

215 IZN Seminar Series

Kelly Rogers, Institut Louis Pasteur, Paris Hans-Rudolf Brenner, Biozentrum Basel Visualisation of local Ca2+ dynamics with genetically en- Neural control of synapse-specific gene expression at the coded bioluminescent reporters neuromuscular junction Eckehard Freitag, Universität Leipzig Roger Traub, Dept. of Physiology, Pharmacology and Neurology, Laser-mediated analysis of cellular events - Netrin-1 overex- State University New York/USA pression studies on neural cells Evidence for electrical coupling between the axons of pyramidal neurons, and the role of such coupling in generating Seminars Clinical Neurobiology oscillations Samuel Lagier, Institut Pasteur, Paris 2003-2005 Gamma oscillations in the rodent olfactory bulb: generation mechanisms and function Rossella Conti, Laboratoire de Physiologie Cérébrale, Université Wolfgang Driever, University of Freiburg de Paris, France Developmental neurogenetics in zebrafish: Cell specification Two-photon imaging of Action Potential and ryanodine in the dopaminergic and noradrenergic systems evoked Ca2+ transients in synaptic terminals of cerebellar interneurons Tatsuhiro Hisatsune, Dept. Integrated Biosciences, University of Tokyo Sebastian Jungnickel, Howard Florey Institute for Experimental Theta oscillations activate adult hippocampal stem cells Physiology and Medicine, Melbourne, Australia Role of neuropeptides in the motor/sensory function of the inferior olive-cerebellum: Galanin systems in the mouse Elisabeth Foeller, Zoologisches Institut, Uni Frankfurt A possible role of inhibition in whisker map plasticity in rat somatosensory cortex Gernot Riedel, University of Aberdeen Involvement of dentate gyrus in memory formation in mice Tatiana Korotkova, Institut für Neurophysiologie, Universität Düsseldorf Hypothalamic modulation of the midbrain dopaminergic system Alexei Ponomarenko, Institut für Neurophysiologie, Universität Düsseldorf The why, where and how of hippocampal ripple oscillations Nail Burnashev, Dept. of Experimental Neurophysiology, Frije Universiteit Amsterdam Smart mouse: NR2B or not NR2B? Dr. K. V. Raghavan, National Centre for Biological Sciences, Ban- galore, India Muscle identity, fibre number, and the development of movement in Drosophila Michael Reid, Rutgers University, New York Spiral ganglion neurons; not so simple after all Olivia Dumitrescu, MPI f. Hirnforschung, Frankfurt Glutamate receptors of amacrine cells in the mouse retina

216 Symposia

IZN Retreat/ SFB 488/ FOR 302 Symposium IZN Retreat Kloster Schöntal, June 1-3, 2004 Kloster Schöntal, June 25-26, 2003 “Development & Circuit Formation” Introduction: Klaus Unsicker Session 1: Session 1: Chair: Hannah Monyer Chair: Klaus Unsicker Laure Bally-Cuif, Neuherberg Erwin Neher Dynamics of neurogenesis at the zebrafish midbrain-hind- Modulation of short-term synaptic plasticity brain boundary Yehezkel Ben-Ari Claudio Stern, London GABA: a pioneer transmitter Molecular dissection of neural induction in the chick embryo Rüdiger Klein, Munich Session2: Role of ephrins in neuronal networking and plasticity Chair: Hilmar Bading Andrew Lumsden Session 2: Forebrain patterning – a new perspective Chair: Andreas Draguhn Diethelm Richter Eric Frank, Boston Serotonin receptors: guardians of stable breathing The beginning of a molecular basis for the formation of the simple stretch reflex Session 3: Javier de Felipe, Madrid Chair: Kerry Tucker Cortical interneurons: From Cajal to 2003 Mart Saarma Structure and biology of GDNF family neurotrophic factor Session 3: receptors Chair: Andreas Draguhn Carlos Ibanez Manfred Heckmann, Freiburg Trophic signaling in the nervous system Electrical events and exocytosis in mossy fiber Christopher Henderson Henry Markram, Lausanne Coordinated development of individual motor units Molecular basis of electrophysiological diversity Session 3: Session 4: Chair: Andreas Draguhn Chair: Hilmar Bading Stephen P. Hunt, London Round table: Substance P in depression and addiction Goals and strategies of European Neuroscience Centers Thursday, June 3 Christian Büchel, Hamburg The importance of connectivity for brain function Session 5: Chair: Joachim Kirsch Session 5: Chair: Hannah Monyer Anders Björklund Induction of Parkinson-like neurodegeneration by overex- Herta Flor, Mannheim pression of human alpha-synuclein in the nigrostriatal sys- Learning, neuroplasticity and psychopathology tem

217 Symposia

Nils Brose Session 3: Short term synaptic plasticity: Molecules and mechanisms Chair: A. Draguhn

Session 6: Wolfgang Wurst Chair: Ulrich Misgeld Dissection of the genetic pathway controlling midbrain dopaminergic neuron induction and differentiation Olaf Pongs Susan Ackerman On the episodic nature of channelopathies Genetic control of cerebellar development Kai Kaila GABA: an exciting inhibitory transmitter Session 4: Chair: G. E. Pollerberg Session 7: Chair: Christoph Schuster Mary Hatten New directions in CNS migration Thomas Gasser Elke Stein Genetics of PD: Monogenic forms as models for the disease Molecular mechanism of axon guidance in the developing Lars Olson vertebrate CNS Genetic risk factors in Parkinson’s disease Esther Stoeckli New tricks for an old dog: Sonic hedgehog is a guidance cue for postcommissural axons SFB 488 Symposium / IZN Retreat Kloster Schöntal, June 15-17, 2005 Session 5: Chair: H. Simon “Neural Development: From Patterning to Network Formation” Liliana Minichiello TrkB regulates neocortex formation through the Shc/PLCg- mediated control of neuronal migration Session 1: Dennis O’Leary Chair: H. Bading Genetic specification of cortical areas and implications for behaviour Yoshiki Sasai Molecular and cellular control of regional specification in the Session 6: nervous system Chair: K. Unsicker Wieland Huttner Tom Curran Cell biology of neurogenesis The Shh pathway and pediatric brain tumors Session 2: Hermann Rohrer Chair: K. Tucker Specification and differentiation of autonomic neurons

Guo-Ping Fan Session 7: Epigenetic gene regulation in neural development Chair: C.Schuster Patrizia Casaccia-Bonnefil Patrik Ernfors Histone deacetylases in oligodendrocyte development The transcriptional control of nociceptive neuron subtype specification Frances Lefcort Imaging formation of the DRG and sympathetic ganglia

218 Symposia

Session 8: Session 5: Chair: U. Misgeld Chair: U. Misgeld Qiufu Ma Alexander Dityatev, Hamburg Molecular control of nociceptive/pain sensory neuron deve- Recognition and neurotransmission: impact of recognition lopment molecules on development of glutamatergic and GABAergic systems in the hippocampus

SFB 488 / FOR 302 Symposium / IZN Retreat Session 6: Chair: H. Monyer Kloster Schöntal, July 2-4, 2006 Josef Bischofberger, Freiburg ‘Development of Transmitter Systems’ Associative synaptic plasticity in mature and newly generated hippocampal granule cells Session 1: Hans Van Hooft, Amsterdam Chair: H. Bading Serotonergic control of postnatal cortical development by 5-HT3 receptor Susanne Schoch, Bonn Genetic ablation of alpha-RIMs: effect on synaptic functions Session 7: Jens Rettig, Homburg Chair: K. Unsicker The role of CAPS1 in LDCV secretion from adrenal chromaf- Chris Deppmann, Baltimore fin cells Neurotrophin Signaling and the Development of the Sympa- thetic Nervous System Session 2: Chair: K. Tucker Marten Smidt, Utrecht Development and subset specification of the mesodience- Nicholas Spitzer, La Jolla phalic dopaminergic system Activity-dependent transmitter specification and receptor matching Session 8: Chair: C. Schuster Session 3: Chair: G.E. Pollerberg Roland Friedel, Munich Role of Plexins and Semaphorins in cerebellar development Tania Vitalis, Paris Amir Dori, Tel-Aviv Origin and fate determination of a subpopulation of telence- Acetylcholinesterase splice variants influence murine neo- phalic interneurons cortical development through catalytic and non-catalytic Stewart Anderson, New York mechanisms Specification of cortical interneurons

Session 4: Chair: A. Draguhn Claudio Rivera, Helsinki Role of the neuron specific K-Cl cotransporter KCC2 in synaptic transmission during development and trauma Derek Sieburth, Boston Systematic analysis of neurotransmission in C. elegans

219 Symposia

IZN/SFB 488 Retreat IZN/SFB 488 Retreat Kloster Schöntal, July 8-9, 2007 Kloster Schöntal, July 20-21, 2008 “Neuronal Networks: from Cell Biology to Cognition?” “Technologies for the Neurosciences”

Opening Lecture: Session 1: Chair: J. Wittbrodt Stephan Frings Olfactory receptor neurons - feeding a sensory network Johann Engelhardt, DKFZ High resolution fluorescence microscopy beyond the diffrac- Highlight Lecture: tion limit Thomas Söllner Ernst Stelzer, EMBL Molecular machinery involved in regulated exocytosis Light sheet based fluorescence microscopes (LSFM, SPIM, DSLM) reduce phototoxic effects by several orders of mag- Session 1: nitude Chair: K. Unsicker Special Lecture: Rohini Kuner Chair: H. Bading The cannabinoid system and pain: novel insights via molecular and genetic approaches Angela Stevens, Applied Mathematics, Uni HD Mathematical modeling and analysis in the life-sciences: Ex- Ulrike Müller amples and perspectives The APP gene family: genetic models to dissect functional domains Session 2: Chair: G. Schratt Session 2: Chair: C. Schuster Holger Erfle, EMBL/BioQuant RNAi screening by automated microscopy Hartwig Spors Spatio-temporal patterns in the mammalian brain - in vivo Joe Lewis, EMBL imaging with voltage sensitive and calcium sensitive dyes The Chemical Biology Core Facility: finding needles in haystacks Christian Fiebach Session 3: Neural networks of verbal working memory: FMRI studies Chair: A. Draguhn Session 3: Moritz Helmstädter, MPIMF Chair: S. Frings Reconstructing neural circuits using serial blockface Scan- ning Electron Microscopy and machine learning Detlev Arendt Evolution of the telencephalon: associative brain centres in a Dirk-Peter Herten, BioQuant marine annelid, Platynereis dumerilii Single-molecule spectroscopy in living cells Rainer Spanagel Session 4: Clock genes and their role in psychopathology Chair: G. Köhr Thomas Kuner Synaptic inhibition accelerates odor discrimination in mice Ulrich Schwarz, BIOMS Modelling the stochastic dynamics of adhesion sites Fred Hamprecht, IWR Pattern recognition and image processing

220 Symposia

SFB488 Symposium Liqun Luo Exploring neural circuits using genetic mosaic methods in Heidelberg, October 14-16, 2004 flies and mice “Developmental Neurobiology: Matthias Landgraf From Molecules to Neural Systems” Patterning of dendrites in the Drosophila CNS

Session 5: Synapse Formation Session 1: Mechanisms of Neural Crest Development Chair: Andreas Draguhn Chair: Klaus Unsicker Peter Scheiffele Nicole Le Doarin Control of CNS synapse formation by the Neuroligin-Neure- The neural crest in the development of the vertebrate head xin adhesion system Marianne Bronner-Fraser Eckart Gundelfinger Hierarchy of regulatory events in neural crest formation Assembly and molecular organization of the active zone of neurotransmitter release Session 2: Wnt Signaling and Neural Crest Development Pico Caroni Chair: Jochen Wittbrodt Mechanisms controlling synapse maintenance and remodelling in young adults Chaya Kalcheim Regulation of neural crest delamination by BMP-dependent Patricia Salinas Wnt signalling Regulation of neuronal connectivity: a role for Wnt signalling Andy McMahon Session 6: Development of Neural Systems Ligand export and feedback control in HH-mediated patter- Chair: Ulrich Misgeld ning of the mammalian neural tube David Raible Peter Mombaerts Reiterated Wnt signaling in zebrafish neural crest develop- Olfaction targeted ment Wenbiao Gan Lukas Sommer Synaptic plasticity and pathology in vivo Wnt signaling in early neural crest stem cells Miles Whittington The rise and fall of gamma rhythms: Oscillations in the deve- Session 3: Neuronal Stem Cells loping and ageing brain Chair: Hilmar Bading Miriam Bibel Differentiation of mouse embryonic stem cells in Drosophila SFB 488 Symposium Magdalena Götz Heidelberg, September 20-23, 2007 Neurogenesis from glial cells: Pax6 as a master regulator? “TGF-ßs: Signaling and Roles in Neural Development, Session 4: Neuron Survival and Shape Maintenance, and Disease” Chair: Joachim Kirsch

Christopher Cowan Session 1: Ephs and GEFs: Coupling receptor activation to endocytosis, Chair: Klaus Unsicker axon guidance and synapse formation Michael B. Sporn, Hanover Alex Kolodkin New triterpenoid drugs that enhance TGF-beta signaling Molecular mechanisms of neuronal growth cone guidance and are neuroprotective

221 Symposia

Rik Derynck, San Francisco Session 6: Non-Smad signaling pathways complement Smad-media- Chair: Mart Saarma ted signaling by TGF-ß Kristján R. Jessen, London Session 2: The role of TGF beta in perinatal nerves Chair: Michael B. Sporn Vassilis Pachnis, London The Ret signaling pathway and its role in enteric nervous sys- Carl-Henrik Heldin, Uppsala tem development Signaling via the TGF-beta receptors: possible targets in tumor treatment Session 7: Peter ten Dijke, Leiden Chair: Kerstin Krieglstein TGF-b signaling in vascular development and diseases Thomas Holstein, Heidelberg Session 3: BMP/Chordin signaling and Cnidarian neurogenesis Chair: Jochen Wittbrodt Mart Saarma, Helsinki Christof Niehrs, Heidelberg GDNF interactions with old and new receptors Regulation of BMP signalling Edward de Robertis, Los Angeles Session 8: A self-regulating system of TGFβ/BMP cell-cell signaling that Chair: Tony Wyss-Coray integrates embryonic patterning Carlos Ibanez, Stockholm TGFß superfamily signaling in the nervous system Session 4: Chair: Christof Niehrs Kerstin Krieglstein, Göttingen Role of TGF-ß in nervous system development Petra Knaus, Berlin Stuart E. Dryer, Houston Regulation of BMP signaling by receptor endocytosis and TGF-ß signaling and the regulation of ion channel trafficking receptor associated proteins in developing neurons Joachim Wittbrodt, Heidelberg Factors and signals in eye development and morphogenesis Session 9: Chaya Kalcheim, Jerusalem Chair: Michael Sendtner A BMP-dependent molecular cascade that generates neural Brian McCabe, New York crest cell migration Neuronal activity and retrograde BMP signaling at the synapse Session 5: Chair: Chaya Kalcheim Michael B. O’Connor, Minneapolis Role of BMP signaling in hippocampal mediated learning Lukas Sommer, Zürich and memory in the mouse TGFbeta signaling regulating proliferation and fate decisions Tony Wyss-Coray, Stanford in neural stem cells Genetic mouse models for the study of TGF-beta signaling in Danny Huylebroeck, Leuven the brain Sip1/Zfhx1b in CNS and PNS development and Mowat-Wil- son syndrome Session 10: Suzana Atanasoski, Basel Chair: Thomas Holstein The role of Ski in neural development Adriano Fontana, Zürich On the role of TGFbeta in infectious and autoimmune diseases of the nervous system

222 Symposia

Ludwig Aigner, Regensburg Session 2: Regulation of neurogenesis in the healthy and diseased Chair: Peter Seeburg brain: a potential role of TGF-beta1 Joachim Kirsch Session 11: Targeting inhibitory synapses: How neurotransmitter recep- Chair: Adriano Fontana tors find their ways William Wisden Michael Weller, Tübingen GABA-A receptors in brain development TGF-ß: a key mediator of the malignant phenotype of glioblastoma Dirk Feldmeyer Early postnatal development in the Barrel Cortex Ulrich Bogdahn, Regensburg TGF-ß based therapy of malignant glioma (from bench to Session 3: bedside) Chair: Hannah Monyer Patrick Mehlen, Lyon The dependence Receptor notion: when apoptosis controls Stefan Offermanns nervous system development and tumorigenesis Gq/11 and G12/13 functions in the nervous system Rohini Kuner Session 12: Expression and functions of plexin- B family members in the Chair: Ulrich Bogdahn developing rodent nervous system Michael Sendtner, Würzburg Dusan Barsch Mouse models of motorneuron disease Conditional regulation of minibrain kinase expression Klaus Unsicker, Heidelberg Peter Seeburg Functions of GDF-15 in the nervous system Behavioural consequences in mice from GnRH promoter- directed changes in AMPA receptors

SFB 488 “Christmas Symposia“ 2003 SFB 488 “Christmas Symposia“ 2004

Session 1: Session 1: Chair: Klaus Unsicker Chair: Peter Seeburg Christof Niehrs Christof Niehrs The transmembrane protein FLRT3 is a novel regulator of Large scale microarray analysis provides a panoramic view of FGF signalling embryonic gene expression Jochen Wittbrodt Klaus Unsicker Control of proliferation and differentiation in the developing The sympathoadrenal cell lineage: adrenal cortex is not re- retina quired for chromaffin cell specification Uwe Strähle Hans-Hermann Gerdes Regulation of the neural determination gene neurogenin1 Tunneling nanotubes: a new principle of intercellular com- in the zebrafish CNS munication for neuronal cells? Andreas Draguhn G. Elisabeth Pollerberg Function and plasticity of synaptic inhibition – the role of Cell adhesion molecule DM-GRASP: Glue or cue? GABA-synthesis and GABA-uptake

223 Symposia

Session 2: Session 2: Chair: Andreas Draguhn Chair. Stefan Offermanns Ulrich Misgeld Hannah Monyer Modulation of inhibition in substantia nigra by retrograde Neurogenesis of distinct GABAergic interneurons in the signals postnatal brain Hannah Monyer Ulrich Misgeld GAP junctions and their role in network synchrony GABAergic control of retrograde signaling in substantia nigra Veit Witzemann Joachim Kirsch Control of synapse formation in muscle Map4K4 - a novel regulator of glycine receptor clustering?

Session 3: Session 3: Chair: Klaus Unsicker Chair: Klaus Unsicker Günther Schütz Thomas Holstein Analysis of glucocorticoid and mineralocorticoid receptor Wnt-signalling and cnidarian neurogenesis function by gene targeting Christof Niehrs Hilmar Bading The role of Kremen in Wnt modulation during neural crest Nuclear calcium signalling in plasticity and neuronal survi- specification val Rohini Kuner Andreas Draguhn Plexin-B2 modulates neural patterning in vivo GABA-metabolism and the plasticity of GABAergic synapses Peter Seeburg SFB 488 “Christmas Symposia“ 2006 Tet-regulated Cre in hypothalamic GnRH neurons of the mouse Session 1: Chair: Hilmar Bading SFB 488 “Christmas Symposia“ 2005 Gerhard Schratt MicroRNAs in synapse development Session 1: Chair: Hilmar Bading Veit Witzemann Formation of neuromuscular junctions: neurocentric versus Gerhard Schratt myogenic view MicroRNAs in synapse development Rainer Friedrich Veit Witzemann Function and development of neuronal circuits in the zebra- Formation of neuromuscular junctions: neurocentric versus fish olfactory bulb myogenic view Peter Seeburg Rainer Friedrich Gene regulation in defined hypothalamic cell populations Function and development of neuronal circuits in the zebrafish olfactory bulb Session 2: Peter Seeburg Chair: Stefan Offermanns Gene regulation in defined hypothalamic cell populations Hannah Monyer Neurogenesis of distinct GABAergic interneurons in the postnatal brain Ulrich Misgeld GABAergic control of retrograde signaling in substantia nigra

224 Symposia

Joachim Kirsch Darren Gilmour Map4K4 – a novel regulator of glycine receptor clustering? Coordinating cell movement and shape within neurogenic placodes Session 3: Chair: Klaus Unsicker Session 2: Chair: Stefan Offermanns Thomas Holstein Wnt-signalling and cnidarian neurogenesis Rohini Kuner Christoph Niehrs Role of Plexin-B proteins in laminar development of the cor- The role of Kremen in Wnt modulation during neural crest tex and spinal cord specification Veit Witzemann Rohini Kuner AChR-mediated activity determines positioning of synapses Plexin-B2 modulates neural patterning in vivo and nerve growth Ingrid Lohmann SFB 488 “Christmas Symposia“ 2007 Transcriptional control of morphogenesis by Hox proteins Laurence Ettwiller Session 1: Highly conserved non-coding DNA elements in fish reveal Chair: Jochen Wittbrodt enhancer function in neuronal structures

Christof Niehrs Session 3: DNA demethylation, DNA repair and pluripotency Chair: Andreas Draguhn Thomas Holstein Structure and function of an unusual neuronal cell type Peter Seeburg Genetic manipulation of hypothalamic functions Christoph Schuster Flies and Psychiatry? Thomas Dresbach (Kirsch group) Role of Neuroligins in presynaptic maturation Session 2: Christoph Schuster Chair: Klaus Unsicker Myosin-VIIa is required to maintain postsynaptic glutamate receptor function Thorsten Bus GnRH neuron specific gene manipulation Joachim Kirsch “Recent advances and perspectives in Glycine receptors in spinal cord development developmental biology” Andreas Draguhn Heidelberg, October 24-28, 2007 Assembly formation and memory consolidation in the hippocampus A trilateral meeting organized by the Hebrew University, Jerusalem, and its partners, the Universities of Göttin- SFB 488 “Christmas Symposia“ 2008 gen and Heidelberg

Session 1 : Chair: Jochen Wittbrodt Keynote Lecture Chair: Klaus Unsicker Thomas Holstein Comparative genomics and the origin of the nervous system Chaya Kalcheim Detlev Arendt Lineage segregation in the somitic mesoderm Evolution of the central nervous system in animals

225 Symposia

Session 2: Stem cells, neurogenesis Session 6: Neuron survival and death II Chair: Chaya Kalcheim Chair: Kerstin Krieglstein Andreas Wodarz Offer Gerlitz Molecular control of cell polarity and asymmetric cell divisi- S149 is a new Dpp target gene that acts as a co-repressor on in Drosophila with Brinker to promote cell death Victor Tarabykin Francesca Ciccolini Transcriptional control of cerebral cortex development Regulation of stem cell proliferation in the postnatal subventricular zone Session 3: Gastrulation and signalling pathways Chair: Kerry Tucker Session 7: Neuron survival and death II Chair: Jochen Wittbrodt Zeev Paroush Groucho-mediated transcriptional repression in Drosophila Günther Schütz development Regulation of dopaminergic neuron survival Christof Niehrs Kerstin Krieglstein New aspects of neural crest development In vivo cooperativity of GDNF and TGFß in the regulation of neuron survival Session 4: Cell migration, Axon guidance I Chair: Zeev Paroush Session 8: Development of epi- and endodermal derivatives Chair: Christof Niehrs Joel Yisraeli The role of VICKZ proteins in cell migration and metastasis Uri Gat Kerry Tucker The Runx family: from hair development in mice to the sea- ENU screen for axonal pathfinding errors anemone Nematostella Avihu Klar Tomas Pieler Molecular mechanisms for increasing complexity of axonal Pancreas development in Xenopus laevis guidance cues Annette Borchers The function of PTK7 in neural crest migration Session 5: Axon guidance II, Synaptogenesis Thomas Holstein Chair: Avihu Klar Patterning of an ancient nervous system Gudrun Rappold Cell motility and mental retardation Session 9: ..and back to very early phylogenesis and ontogenesis. Achim Kirsch Chair: Klaus Unsicker Glycine receptor development Herbert Steinbeisser Session 6: Neuron survival and death I Gastrulation in Xenopus Chair: Kerstin Krieglstein Jochen Wittbrodt Oded Behar Morphogenesis of the optic cup Semaphorins and neurotrophins in axon guidance and cell death

226 Symposia

Joint Symposium on Neurocomputing Yosef Yarom June 16-17, 2003 Oscillating without gap junctions David Hanse Center for Molecular Biology, ZMBH Electrical synapses and synchrony: The role of intrinsic currents Hannah Monyer Scientific Session I: New Microscopy Methods The oscillating brain approached by mouse mutants Chair: K. Unsicker Jörg Langowski Fluorescence fluctuation microscopy as a tool for under- 1. Bioquant Symposium standing molecular motion in cells December 9, 2004 Christoph Cremer New microscopy methods to study cell nuclear structure and its relation to gene regulation Opening Remarks: The Bioquant concept Winfried Denk Jochen Tröger, Willi Jäger, Bernd Bukau, Angret Joester Direction selectivity in the retina Session 1: Modeling in the life sciences Scientific Session II: Modeling Chair: Roland Eils Chair: A. Lewis Joachim Spatz, MPI für Metallforschung - Stuttgart und Biophysi- Tali Tishby kalische Chemie, Universität Heidelberg Quantitative principles of biological information processing Models for studying chemomechanical coupling in cell adhesion and development Idan Segev Cooperative synaptic plasticity in cortical dendrites Matthias Weiß, MEMPHYS - Center for biomembrane physics, Odense, Denmark und BIOMS, Heidelberg Rainer Dalhaus Structure formation of active biomembranes On the identification of synaptic connections in neural networks by graphical models Ulrich Schwarz, MPI für Kolloid- und Grenzflächenforschung, Potsdam und BIOMS, Heidelberg Andre Rupp Towards a quantitative understanding of cell adhesion Cortical processing of auditory information Gabriel Wittum, Interdisziplinäres Zentrum für Wissenschaftli- Israel Nelken ches Rechnen, Heidelberg Transformations of stimulus representation in the ascending Towards Simulation of Neuronal Signal Processing auditory system Session 2: Technologies for systems biology Scientific Session III: Cortical Dynamics Chair: Christoph Cremer Chair: I. Segev Irmgard Sinning, Biochemiezentrum, Heidelberg Aaron Lewis Quantitative analysis of SRP receptor-membrane interac- Functional imaging of neurons and neural networks tion Michael Brecht Stefan Hell, MPI für biophysikalische Chemie, Göttingen und Sensorimotor representations and their cellular codes in the DKFZ, Heidelberg rat necortex Fluorescence nanoscopy: Concepts, experiments and instru- Rony Paz (Lab E. Vaadia) ments Improvement of movements representation in the motor cortex of a monkey during learning

227 Symposia

Keynote lecture GK 791 Retreat Chair: Willi Jäger Odenwald, October 24-25, 2003 Hans Westerhoff, Vrije Universiteit, Amsterdam Integrative systems biology: Bringing genomes to life Friday, december 10 Session I Julia Mack (Dr. Rainer Friedrich, MPI, Heidelberg) Session 3: Functional optical analysis of inhibitory interneurons in the Challenges for systems biology in plant science and medicine developing olfactory bulb of zebrafish Chair: Klaus Unsicker Christine Hassler ( Dr. Christoph Niehrs, DKFZ, Heidelberg) Rüdiger Hell, Heidelberger Institut für Pflanzenwissenschaften The mechanism of axis formation in the developing embryo Heidelberg Caroline Schmitz ( Dr. Harald Hutter, MPI, Heidelberg) Functional characterization of sulfur metabolite-based regu- Screens for genes controlling axon guidance in C. elegans latory networks in plants Benedikt Kost, Heidelberger Institut für Pflanzenwissenschaften, Session II Heidelberg Characterization of the actomyosin system mediating orga- Bettina Maier ( Prof. Dr. Elisabeth Pollerberg, Institute of Zoology, nelle motility in pollen tubes Dept. of Developmental Neurobiology, Heidelberg) Functional analysis of axonal cell adhesion molecules (CAMs) Pascal Tomakidi, Poliklinik für Kieferorthopädie, Heidelberg manipulated in their expression levels in the developing vi- Organotypic co-cultures: Tools to study morphogenesis of sual system periodontal tissues in vitro Suhua Deng ( Prof. Dr. Stefan Offermanns, Institute of Pharmaco- Christiane Schönbein (AG Mahlknecht), Medizinische Klinik und logy Heidelberg) Poliklinik V, Heidelberg Generation and analysis of mice lacking the plexin-B family Analysis of epigenetical effects on the differentiation of he- members plexin-B1 and plexing-B2 matopoietic stem cells Session 4: Modeling of signal transduction Session III Chair: Thomas Rausch Gitta Erdmann ( Prof. Dr. Günther Schütz, DKFZ, Heidelberg) Victor Sourjik, ZMBH, Heidelberg Analysis of glucocorticoid receptor Signal processing in bacterial chemotaxis Claus Beck ( Prof. Dr. Dusan Bartsch, Zentralinstitut für seelische Stephan Frings, Zoologie, Heidelberg Gesundheit – Mannheim) Olfactory sensory cilia – from cartoon to function Down syndrome and cognitive processes: The Minibrain Kinase Ursula Klingmüller, DKFZ, Heidelberg Nidhi Gakhar ( Dr. Francesca Ciccolini, Prof. Dr. Hilmar Bading, Dynamic modelling of the JAK-STAT signaling pathway Neurobiology-Heidelberg) Rainer Friedrich, MPI für medizinische Forschung, Heidelberg The role of Calcium in embryonic striatal neural precursors Neuronal circuits in the olfactory system: neurophysiology Markus Uhrig ( Dr. Tobias Hartmann, DKFZ, Heidelberg) and computation Gene expression profiling of human neuroblastoma cells Peter Bengtson, Interdisziplinäres Zentrum für Neurowissen- exposed to Ab42 and neural differentiation of human adult schaften, Heidelberg stem cells in vitro and in vivo Transcription-dependent plasticity: Input/output function of nuclear calcium signaling in hippocampal neurons Session IV

Strategic discussion (group leaders only) Ayla Arslan ( Dr. William Wisden, Clinical Neurobiology Heidelberg) Chairs: Winfried Denk, Roland Eils A receptors segregation of different subunits in the meme- brane and modulation of benzodiazepine pharmacology for studying specific neuronal circuits

228 Symposia

Oliver Schlicker ( PD Dr. Hans-Hermann Gerdes, Neurobiology Markus Uhrig, ZMBH, Dr. Tobias Hartmann Heidelberg) Gene expression profiling of human neuroblastoma cells over- GABAA-receptors in dendritic membrane traffic expressing A peptides in the context of Alzheimer’s disease Beril Doganci ( Prof. Dr. Hannah Monyer, Clinical Neurobiology Heidelberg) Sabine Chourbaji, Central Institute of Mental Health (CIMH), NR2B conditional knock-out Mannheim, PD Dr. Peter Gass/ Prof. Dr. Fritz Henn The significance of multiple stressors in prehistory for the Session V specificity in the learned helplessness model of depression in mice Aleksandar Zivkovic ( Dr. Andrei Rozov, Clinical Neurobiology Heidelberg) Claus Beck, Central Institute for Mental Health Mannheim, Prof. Characterization of excitatory synaptic transmission in GluR- Dr. Dusan Bartsch D knock-out mice Minimal Animal model for cognitive deficits in Down Syn- drome Dragos Inta ( Prof. Dr. Hannah Monyer, Clinical Neurobiology Hei- delberg) Session II Generation of transgenic mice expressing enhanced green fluorescent protein (EGFP) in 5-HT3 receptor positive neu- Caroline Schmitz, MPI für medizinische Forschung, Dr. H. Hutter rons Identification of genes controlling axon guidance in C. ele- Srinivasa Subramaniam ( Prof. Dr. Klaus Unsicker, Neuroanatomy gans by a large scale RNAi screen Heidelberg) Bettina Maier, Institute of Zoology, Dept. of Developmental Neu- ERK activation promotes degeneration of cerebellar granule robiology, Prof. G .E. Pollerberg neurons independent of caspase-3 Functional analysis of axonal cell adhesion molecules (CAMs) Ioana Potrovita ( PD Dr. Markus Schwaninger, Neurology Heidel- manipulated in their expression levels in the developing vi- berg) sual system TWEAK a new protein of the TNFk family and its role on cere- Suhua Deng, Institute of Pharmacology, University of Heidelberg, bral ischemia Professor Stefan Offermanns Generation and analysis of mice lacking the plexin-B family Session VI members plexin-B1 and plexin-B2 Corina Popovici ( Prof. Dr. Klaus Unsicker, Neuroanatomy Heidel- Julia Mack, MPI für medizinische Forschung, Dr. Rainer Friedrich berg) Early development of functional spatial maps in the zebrafish olfactory bulb Sabine Chourbaji ( Prof. Dr. Fritz A. Henn, Zentralinstitut für seelische Gesundheit, Mannheim) Session III Behavioural aspects of animal models for depressive disorders Dragos Inta, Clinical Neurobiology, Professor Dr. Hannah Monyer Expression profile of 5-HT3-positive neurons in a transgenic mouse model GK 791 Retreat Ioana Inta, Neurology, AG. PD. Dr. Markus Schwaninger March 18-19, 2005 TWEAK and NF-kB in cerebral ischemia Nidhi Gakhar, Neurobiology, Prof. Dr. Hilmar Bading Calcium sgnalling in nural pecursors Session I

Gitta Erdmann, DKFZ, Prof. Günther Schütz Session IV Generating a mouse with a mutated glucocorticoid receptor allele leading to an impaired nulcear import Aleksandar Zikovic, Clinical Neurobiology, Dr. Andrei Rozov Role of Homer 1a in termination of LTP

229 Symposia

Ali Cetin, MPI, Department of Molecular Neurobiology, Prof. Dr. Peter Seeburg Lentiviral based approach to study activity dependent development of cortical networks in the rat barrel cortex Oliver Schlicker, Dept. of Neurobiology, Prof. Dr. Hans-Hermann Gerdes GABAA-rceptors in dendritic membrane traffic Beril Doganci, Clinical Neurobiology, Prof. Dr. Hannah Monyer Functional analysis of NR2B subunit

230 Symposia

231 Map of Campus

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720

37 P HTC 584 Berliner Straße 701 - 703 583 Technologiepark 700 585 531 582 520 Fernheizwerk 581 530 Technologiepark 580 705 509- 515 P 704 671 P 706 Pädagogische 506 524 Hochschule Max-PIanck- 523 562 678 VZM Institut 696 P 535 505 522

683 501-504 P 521 37 694 670 536 P 680 P 560 695 677 693 681 561 Ü P 21, 24 684 Studenten- 692 wohnheime 676 672 31 254 31, 37 691 685 687 675 294 293 235 31, 37, 721 Geogr. Institut 686 682 690 236 (Berliner Str. 48) 31, 32, 37 292 253 234 SAI 276 252 P 689 688 P 269 233 31 P 350 330 274 232 TSG 277 231 Ü 450 Kopf- OMZ 272 278 267 230 HIT klinik P 430 368 348 275 268 400 270 229a 328 T 308 273

i 347a e r 271 229 g Neue 367 a 347 Ü 31 r 327 307 t e Kinderklinik 288 n s THEORETIKUM t 31 r a 366 306 31 ß 346 326 21, 24 e Medizinische P Klinik 345 227 365 325 305 P P 364 324 410 Mensa 425 344a 304 344b 242 Botanischer Garten 282 201 31 222 163 360 281 261 241

370 Straße Berliner Zoo 162 31, 32 336-44 223 280 260 220 P 160 240 154 152 DKFZ 221 Nieren- 153 161 151 320 zentrum 155 P 371 31, 32 110 100 163 156 P Kinderklinik P 31,32 UBA 111 105 114 133 P 115 Ü 159 21, 24 Reiterverein 131 IZN Investigators 130 119 116 31, 32 132 P Universität/Klinikum 134 112 Max-PIanck-Institut 129 (Jahnstr. 29) Univ./Klinikum (im Bau) Chirurgische Klinik THEORETIKUM Sonstige Einrichtungen Wohn-/Gewerbegebiete Hauptbahnhof Klinikum Bergheim Campus Autobahn Altstadt N E C K A R 696 100 m

232 Map of Campus

Campus „Im Neuenheimer Feld“ INF 368 Wittum group Location of IZN Groups INF 400 Rupp group For the most part, the research groups of the IZN Wick group Investigators are accomodated in different buildings on the university campus “Im Neuenheimer Feld” (see INF 581 Niehrs group map, buildings in black). Other IZN groups are located Schütz group in Heidelberg‘s old town, the EMBL, and in Mannheim (Central Institute for Mental Health/ZI, and University MPIMF Denk group Clinics Mannheim). Euler group Köhr group INF 220/221 Von Deimling group Seeburg group Spors group INF 230 Frings group Sprengel group Holstein group Witzemann group Wittbrodt group Groups outside the campus INF 232 Pollerberg group Department of Psychology INF 280 Wiestler group Fiebach group Pauen group INF 282 Kins group EMBL Arendt group INF 307 Ernsberger group Gilmour group Kirsch group Kuner T. group Department of Anesthesiology Mannheim Simon group Schmelz group Tucker group Unsicker group ZI Mannheim Bartsch group Bohus group INF 326 Draguhn group Flor group Gass group INF 345 Schratt group Meyer-Lindenberg group Schuster group Spanagel group Söllner group INF 364 Bading group CBTM Mannheim Mense group Ciccolini group Monyer group Müller group

INF 366 Kuner R. group Rappold group Schwaninger group

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