DOCSLIB.ORG

Advancing Molecular Crustacean Chronobiology Through The

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Advancing Molecular Crustacean Chronobiology Through The Advancing molecular crustacean chronobiology through the characterisation of the circadian clock in two malacostracan species, Euphausia superba and Parhyale hawaiensis Thesis submitted for the degree of Doctor of Philosophy at the University of Leicester by Benjamin James Hunt Department of Genetics University of Leicester 2016 Advancing molecular crustacean chronobiology through the characterisation of the circadian clock in two malacostracan species, Euphausia superba and Parhyale hawaiensis Benjamin James Hunt Abstract The ability to entrain to environmental cycles and therefore anticipate and prepare for the changes they predictably bring is the preserve of the endogenous biological clock, most widely studied at the circadian level. Despite a rich history of research into the behavioural and physiological rhythms shown by many crustacean species, the underlying molecular system driving such traits is not well understood. The aim of this research was to develop our understanding of crustacean clocks through the study of two species, one of major ecological importance and the other a powerful model organism. The Antarctic krill Euphausia superba is a keystone species in the Southern Ocean ecosystem, and evidence suggests that the clock may influence both daily and seasonal rhythms. Using a variety of approaches, including the creation of a de novo assembled head transcriptome, a full suite of clock-related genes have now been cloned and characterised. Unlike many species Euphausia superba possesses orthologs of every canonical core clock gene, and cell culture assays indicate that the central feedback loop has the capacity for complete transcriptional inhibition via two separate pathways, raising the possibility that the krill clock may be an ancestral type or employ multiple oscillators to control rhythms of differing periods. In contrast to the relatively intractable krill, the amphipod Parhyale hawaiensis has simple maintenance requirements and an extensive genetic toolkit with the potential to enable sophisticated dissection of the molecular clock. With the aim of laying the groundwork for future research the clock genes of this species have also been identified, along with the development of a locomotor activity assay. Parhyale hawaiensis shows evidence of bimodal patterns of activity under the control of a molecular clock that combines mammalian-like characteristics with some unique features worthy of further investigation. i Acknowledgements First and foremost my thanks to my supervisor Ezio Rosato, a man of endless and infectious enthusiasm for research who provided me with the perfect level of support and guidance – it has been a pleasure and a privilege to work with him. I am indebted as well to my retired second supervisor Ted Gaten, whose generosity on my return to the fold enabled me to hit the ground, if not running, then at least hobbling slightly faster. Three other people require special thanks. Firstly to my unofficial supervisor Özge Özkaya, who not only laid much of the groundwork for this research but who shouldered the unenviable burden of teaching a ham-fisted zoology graduate the art of molecular benchwork without (visibly) flinching. Secondly to computer wizard Nathaniel Davies, creator of the Euphausia superba Transcriptome Database: Online Edition. Thirdly to Lin Zhang, the other member of lab 124’s tiny crustacean enclave and an ever-reliable source of insight and S2 plasmids. Thank you to every member of our lab past and present, with particular nods to Giorgio Fedele, Celia Hansen, Kam Chana, Jon Shand, Marie Nugent, Marcela Buricova, Ed Green, Laura Delfino, Laura Flavell, Mirko Pegoraro, Elise Facer-Childs and Wilhelmina Assam, all of whom were generous with time, assistance and opinions. Thanks too to the technical and administrative staff, particularly Helen Roe and Liam Gretton, and to Paul Seear for sharing his extensive knowledge. Outside of Leicester, Ana Patricia Ramos and Michalis Averof were generous with their advice and assistance on amphipod wrangling and in their willingness to share data and animals, and am I deeply grateful to Aziz Aboobaker, nastasios Pavlopoulos and Damian Kao for granting me early access to the Parhyale hawaiensis genome. If I tried to fully express my gratitude to family and friends, the acknowledgements section alone would break the 50,000 word limit this thesis is subject to. Mum, dad, granddad, Paula, Chris, Nathan, Manu, Sophie, Isabel and the rest of my family; and everyone who joins me for dinner or drinks from time to time - you have provided support, love and laughs in abundance and I thank you all. Finally, a tribute to two surpassingly wonderful people. Firstly to my fiancée Anna, whose unstinting support has sustained me through this challenge, and others. Even if there was no word limit I could not do you justice - I love you and need you, always. And to my grandma Betty, source of happy childhood memories beyond counting. Gron, I love you too, and I miss you so much. ii Contributors This thesis could not have been produced without the valuable contributions of the following colleagues. Dr. Ted Gaten: Collection and storage of Euphausia superba samples, as described in section 2.2.1.1. Entrainment of Euphausia superba in LD conditions, subsequent collection and storage of samples in LD and DD for use in RT-PCR, as discussed in section 2.4.3.3. Dr. Özge Özkaya: Euphausia superba RNA extraction and storage as described in section 2.2.1.2. Cloning and sequencing of the initial Esbmal1 340 bp fragment through degenerate and semi-degenerate PCR, as detailed in 2.3.1.1. Tissue-specific PCR of Esbmal1, Escry1 and Escry2 (Figure 2.7). Full cloning of Escry1 and Escry2 and characterisation of the former as light-sensitive (Figure 2.13). RT-PCR for Esbmal1, Esclk, Escry1 and Escry2 (2.4.3.3; Figure 2.15). Nathaniel Davies: Production of the Euphausia superba Transcriptome Database, covering the formatting of transcript and annotation data and the production of the website itself. Section 3.2.2.4 details this process in his words. This research used the ALICE High Performance Computing Facility at the University of Leicester and was funded by the National Environmental Research Council (NERC). iii Table of Contents Abstract i Acknowledgements ii Contributors iii Table of Contents iv List of Tables xiii List of Figures xv Common abbreviations and author’s note xix CHAPTER 1 GENERAL INTRODUCTION 1 1.1 The biological clock 1 1.1.1 Ancient rhythms 1 1.1.2 The benefits of a biological clock 1 1.2 The molecular clock 2 1.2.1 The circadian clock in Drosophila melanogaster 2 1.2.2 The core 3 1.2.3 Supporting roles 4 1.2.4 Variations on a theme 5 1.3 Crustacean clocks 5 1.3.1 A history of crustacean chronobiology 5 1.3.2 Molecular data: a long deficit, then a sudden deluge 9 1.4 Euphausia superba 10 1.4.1 Distribution and biomass 10 1.4.2 Life history 11 1.4.3 Ecological and economic importance 11 iv 1.5 The rhythms of Euphausia superba 12 1.5.1 Timekeeping in the Southern Ocean 12 1.5.2 Circadian behaviour 13 1.5.3 Seasonal variations 16 1.6 On the productivity of model and non-model organisms 17 1.6.1 Euphausia superba: high importance, high maintenance 17 1.6.2 The very model of a modern research animal 17 1.6.3 Seeking a pacemaker 19 1.6.4 Parhyale hawaiensis: a model organism par excellence? 22 1.7 Research aims and outline 22 CHAPTER 2 THE CLONING AND CHARACTERISATION OF THE CANONICAL CLOCK GENES OF EUPHAUSIA SUPERBA 24 2.1 Introduction 24 2.1.1 The core genes of the circadian clock 24 2.1.1.1 The central heterodimer 24 2.1.1.2 Repressive elements 26 2.1.1.3 The cryptochromes 27 2.1.1.4 Tying it all together 28 2.1.2 Other organisms 29 2.1.2.1 Insecta 29 2.1.2.2 Crustacea 30 2.1.3 Research aims 30 2.2 Methods 31 2.2.1 Sampling 31 2.2.1.1 Euphausia superba capture and storage 31 2.2.1.2 RNA extraction and cDNA synthesis 31 2.2.2 Gene cloning 32 2.2.2.1 Degenerate PCR 32 2.2.2.2 Searching the SRA 33 2.2.2.3 RACE extension 34 2.2.2.4 Transcriptome mining 34 2.2.2.5 Confirmation 34 2.2.3 Gene characterisation 36 2.2.3.1 Tissue expression 36 v 2.2.3.2 Protein sequence analysis 36 2.2.3.3 Phylogeny 36 2.2.3.4 Transcriptional activity assay 36 2.3 Results 39 2.3.1 Gene cloning 39 2.3.1.1 Esbmal1 39 2.3.1.2 Esclock 39 2.3.1.3 Esperiod 39 2.3.1.4 Estimeless 40 2.3.2 Protein sequence analysis 40 2.3.2.1 EsBMAL1 40 2.3.2.2 EsCLOCK 41 2.3.2.3 EsPERIOD 42 2.3.2.4 EsTIMELESS 42 2.3.2.5 Comparative analysis 43 2.3.3 Tissue expression 44 2.3.4 Phylogenetic trees 48 2.3.4.1 EsBMAL1 48 2.3.4.2 EsCLOCK 48 2.3.4.3 EsPERIOD 48 2.3.4.4 EsTIMELESS 48 2.3.5 Transcriptional activity 53 2.4 Discussion 54 2.4.1 Summary of results 54 2.4.1.1 Esbmal1 54 2.4.1.2 Esclock 54 2.4.1.3 Esperiod 54 2.4.1.4 Estimeless 55 2.4.1.5 The cryptochromes 55 2.4.2 Peptide features and comparisons 56 2.4.2.1 The core heterodimer 56 2.4.2.2 The domains of EsPERIOD and EsTIMELESS 59 2.4.2.3 Evolutionary relationships 60 2.4.3 The molecular clockwork of Euphausia superba 61 2.4.3.1 Transcription, repression and tissue expression 61 2.4.3.2 An ancient clock or a crustacean specialisation? 63 2.4.3.3 Regarding transcript cycling 65 vi 2.4.4 Future work 68 2.4.4.1 Immediate issues 68 2.4.4.2 Following up on a theory 69 2.4.5 Conclusions 70 CHAPTER 3 THE EUPHAUSIA SUPERBA TRANSCRIPTOME DATABASE: DE NOVO ASSEMBLY, ANNOTATION AND OUTPUT.
Load more

Recommended publications
  • A Classification of Living and Fossil Genera of Decapod Crustaceans
    RAFFLES BULLETIN OF ZOOLOGY 2009 Supplement No. 21: 1–109 Date of Publication: 15 Sep.2009 © National University of Singapore A CLASSIFICATION OF LIVING AND FOSSIL GENERA OF DECAPOD CRUSTACEANS Sammy De Grave1, N. Dean Pentcheff 2, Shane T. Ahyong3, Tin-Yam Chan4, Keith A. Crandall5, Peter C. Dworschak6, Darryl L. Felder7, Rodney M. Feldmann8, Charles H. J. M. Fransen9, Laura Y. D. Goulding1, Rafael Lemaitre10, Martyn E. Y. Low11, Joel W. Martin2, Peter K. L. Ng11, Carrie E. Schweitzer12, S. H. Tan11, Dale Tshudy13, Regina Wetzer2 1Oxford University Museum of Natural History, Parks Road, Oxford, OX1 3PW, United Kingdom [email protected] [email protected] 2Natural History Museum of Los Angeles County, 900 Exposition Blvd., Los Angeles, CA 90007 United States of America [email protected] [email protected] [email protected] 3Marine Biodiversity and Biosecurity, NIWA, Private Bag 14901, Kilbirnie Wellington, New Zealand [email protected] 4Institute of Marine Biology, National Taiwan Ocean University, Keelung 20224, Taiwan, Republic of China [email protected] 5Department of Biology and Monte L. Bean Life Science Museum, Brigham Young University, Provo, UT 84602 United States of America [email protected] 6Dritte Zoologische Abteilung, Naturhistorisches Museum, Wien, Austria [email protected] 7Department of Biology, University of Louisiana, Lafayette, LA 70504 United States of America [email protected] 8Department of Geology, Kent State University, Kent, OH 44242 United States of America [email protected] 9Nationaal Natuurhistorisch Museum, P. O. Box 9517, 2300 RA Leiden, The Netherlands [email protected] 10Invertebrate Zoology, Smithsonian Institution, National Museum of Natural History, 10th and Constitution Avenue, Washington, DC 20560 United States of America [email protected] 11Department of Biological Sciences, National University of Singapore, Science Drive 4, Singapore 117543 [email protected] [email protected] [email protected] 12Department of Geology, Kent State University Stark Campus, 6000 Frank Ave.
    [Show full text]
  • New Alien Crayfish Species in Central Europe
    NEW ALIEN CRAYFISH SPECIES IN CENTRAL EUROPE Introduction pathways, life histories, and ecological impacts DISSERTATION zur Erlangung des Doktorgrades Dr. rer. nat. der Fakultät für Naturwissenschaften der Universität Ulm vorgelegt von Christoph Chucholl aus Rosenheim Ulm 2012 NEW ALIEN CRAYFISH SPECIES IN CENTRAL EUROPE Introduction pathways, life histories, and ecological impacts DISSERTATION zur Erlangung des Doktorgrades Dr. rer. nat. der Fakultät für Naturwissenschaften der Universität Ulm vorgelegt von Christoph Chucholl aus Rosenheim Ulm 2012 Amtierender Dekan: Prof. Dr. Axel Groß Erstgutachter: Prof. Dr. Manfred Ayasse Zweitgutachter: Prof. apl. Dr. Gerhard Maier Tag der Prüfung: 16.7.2012 Cover picture: Orconectes immunis male (blue color morph) (photo courtesy of Dr. H. Bellmann) Table of contents Part 1 – Summary Introduction ............................................................................................................................ 1 Invasive alien species – a global menace ....................................................................... 1 “Invasive” matters .......................................................................................................... 2 Crustaceans – successful invaders .................................................................................. 4 The case of alien crayfish in Europe .............................................................................. 5 New versus Old alien crayfish .......................................................................................
    [Show full text]
  • Circadian Clocks in Crustaceans: Identified Neuronal and Cellular Systems
    Circadian clocks in crustaceans: identified neuronal and cellular systems Johannes Strauss, Heinrich Dircksen Department of Zoology, Stockholm University, Svante Arrhenius vag 18A, S-10691 Stockholm, Sweden TABLE OF CONTENTS 1. Abstract 2. Introduction: crustacean circadian biology 2.1. Rhythms and circadian phenomena 2.2. Chronobiological systems in Crustacea 2.3. Pacemakers in crustacean circadian systems 3. The cellular basis of crustacean circadian rhythms 3.1. The retina of the eye 3.1.1. Eye pigment migration and its adaptive role 3.1.2. Receptor potential changes of retinular cells in the electroretinogram (ERG) 3.2. Eyestalk systems and mediators of circadian rhythmicity 3.2.1. Red pigment concentrating hormone (RPCH) 3.2.2. Crustacean hyperglycaemic hormone (CHH) 3.2.3. Pigment-dispersing hormone (PDH) 3.2.4. Serotonin 3.2.5. Melatonin 3.2.6. Further factors with possible effects on circadian rhythmicity 3.3. The caudal photoreceptor of the crayfish terminal abdominal ganglion (CPR) 3.4. Extraretinal brain photoreceptors 3.5. Integration of distributed circadian clock systems and rhythms 4. Comparative aspects of crustacean clocks 4.1. Evolution of circadian pacemakers in arthropods 4.2. Putative clock neurons conserved in crustaceans and insects 4.3. Clock genes in crustaceans 4.3.1. Current knowledge about insect clock genes 4.3.2. Crustacean clock-gene 4.3.3. Crustacean period-gene 4.3.4. Crustacean cryptochrome-gene 5. Perspective 6. Acknowledgements 7. References 1. ABSTRACT Circadian rhythms are known for locomotory and reproductive behaviours, and the functioning of sensory organs, nervous structures, metabolism and developmental processes. The mechanisms and cellular bases of control are mainly inferred from circadian phenomenologies, ablation experiments and pharmacological approaches.
    [Show full text]
  • A Mutant Drosophila Homolog of Mammalian Clock Disrupts
    Cell, Vol. 93, 791±804, May 29, 1998, Copyright 1998 by Cell Press AMutantDrosophila Homolog of Mammalian Clock Disrupts Circadian Rhythms and Transcription of period and timeless Ravi Allada,*²³§ Neal E. White,³ Aronson et al., 1994; Shearman et al., 1997; Sun et al., W. Venus So,²³ Jeffrey C. Hall,²³ 1997; Tei et al., 1997). ²³ and Michael Rosbash* k In Drosophila, there are two well-characterized clock *Howard Hughes Medical Institute genes: period (per) and timeless (tim). Protein levels, ² NSF, Center for Biological Timing RNA levels, and transcription rates of these two genes ³ Department of Biology undergo robust circadian oscillations (Zerr et al., 1990; Brandeis University Hardin et al., 1990, 1992; Hardin, 1994; Sehgal et al., Waltham, Massachusetts 02254 1995; So and Rosbash, 1997). In addition, mutations § Department of Pathology in the two proteins (PER and TIM) alter or abolish the Brigham and Women's Hospital periodicity and phase of these rhythms, demonstrating Boston, Massachusetts 02115 that both proteins regulate their own transcription (Har- din et al., 1990; Sehgal et al., 1995; Marrus et al., 1996). Although there is no evidence indicating that the effects Summary on transcription are direct, PER contains a PAS domain, which has been shown to mediate interactions between We report the identification, characterization, and transcription factors (Huang et al., 1993; Lindebro et cloning of a novel Drosophila circadian rhythm gene, al., 1995). Most of these PAS-containing transcription dClock. The mutant, initially called Jrk, manifests dom- factors also contain the well-characterized basic helix- inant effects: heterozygous flies have a period alter- loop-helix (bHLH) DNA-binding domains (Crews, 1998).
    [Show full text]
  • Case 3:17-Cr-00249-BJD-JRK Document 89 Filed 03/14/18 Page 1 of 32 Pageid 361
    Case 3:17-cr-00249-BJD-JRK Document 89 Filed 03/14/18 Page 1 of 32 PageID 361 UNITED STATES DISTRICT COURT MIDDLE DISTRICT OF FLORIDA JACKSONVILLE DIVISION UNITED STATES OF AMERICA v. CASE NO. 3:17-cr-249-BJD-JRK JOSHUA RYNE GOLDBERG DEFENDANT’S SENTENCING MEMORANDUM The Defendant, Joshua Ryne Goldberg, by and through counsel, Paul Shorstein, Shorstein, Lasnetski & Gihon, LLC, hereby submits the following Sentencing Memorandum setting forth all factors that the Court should consider in determining the type and length of sentence that is sufficient, but not greater than necessary, to comply with the statutory directives set forth in 18 U.S.C. §3553(a). BACKGROUND In 2015, law enforcement officials began investigating a certain online account that was posting threatening and other disturbing comments over the internet. Specifically, some of the internet discussion revolved around a potential attack of a 9/11 event in Kansas City, Missouri. That investigation ultimately led law enforcement officials to the Clay County home of Joshua Goldberg and his family. In the early morning hours of September 10, 2015, FBI agents and other law enforcement officials executed a search warrant inside 1 Case 3:17-cr-00249-BJD-JRK Document 89 Filed 03/14/18 Page 2 of 32 PageID 362 the Goldbergs’ home. They found Joshua Goldberg inside, as he always was. Joshua gave a statement that lasted a few hours despite not having slept for approximately 24 hours and not having taken his medication. Presumably, because the law enforcement officials found a severely mentally ill young man rather than what they expected as the source of the threatening internet activity, the government sought a criminal complaint (case number 3:15-mj-1170-J-JRK), which was filed on September 10, 2015, rather than more formal charges.
    [Show full text]
  • EMERITA TALPOIDA and DONAX VARIABILIS DISTRIBUTION THROUGHOUT CRESCENTIC FORMATIONS; PEA ISLAND NATIONAL WILDLIFE REFUGE a Thesi
    EMERITA TALPOIDA AND DONAX VARIABILIS DISTRIBUTION THROUGHOUT CRESCENTIC FORMATIONS; PEA ISLAND NATIONAL WILDLIFE REFUGE A thesis submitted in partial fulfillment of the requirements for the degree MASTER OF SCIENCE in ENVIRONMENTAL STUDIES by BLAIK PULLEY AUGUST 2008 at THE GRADUATE SCHOOL OF THE COLLEGE OF CHARLESTON Approved by: Dennis Stewart, Thesis Advisor Dr. Robert Dolan Dr. Scott Harris Dr. Lindeke Mills Dr. Amy T. McCandless, Dean of the Graduate School 1454471 1454471 2008 ABSTRACT EMERITA TALPOIDA AND DONAX VARIABILIS DISTRIBUTION THROUGHOUT CRESCENTIC FORMATIONS; PEA ISLAND NATIONAL WILDLIFE REFUGE A thesis submitted in partial fulfillment of the requirements for the degree MASTER OF SCIENCE in ENVIRONMENTAL STUDIES by BLAIK PULLEY JULY 2008 at THE GRADUATE SCHOOL OF THE COLLEGE OF CHARLESTON Pea Island National Wildlife Refuge is a 13-mile stretch of shoreline located on the Outer Banks of North Carolina, 40 miles north of Cape Hatteras and directly south of Oregon Inlet. This Federal Navigation Channel is periodically dredged and sand is placed on the north end of the Pea Island beach. While the sediment nourishes the beach in a particularly sand-starved environment, it also alters the physical and ecological conditions. Most affected are invertebrates living in the swash, the most dominant being the mole crab (Emerita talpoida) and the coquina clam (Donax variabilis). These two species serve as a major food source for shorebirds on the island. It is especially important to protect this food resource on the federal Wildlife Refuge, which operates under a mandate to protect resources for migratory birds. For this research, beach cusps of various sizes were sampled to determine whether there is a correlation between invertebrate populations and the physical characteristics associated with these crescentic features.
    [Show full text]
  • Epigenetic Modifications to Cytosine and Alzheimer's Disease
    University of Kentucky UKnowledge Theses and Dissertations--Chemistry Chemistry 2017 EPIGENETIC MODIFICATIONS TO CYTOSINE AND ALZHEIMER’S DISEASE: A QUANTITATIVE ANALYSIS OF POST-MORTEM TISSUE Elizabeth M. Ellison University of Kentucky, [email protected] Digital Object Identifier: https://doi.org/10.13023/ETD.2017.398 Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Ellison, Elizabeth M., "EPIGENETIC MODIFICATIONS TO CYTOSINE AND ALZHEIMER’S DISEASE: A QUANTITATIVE ANALYSIS OF POST-MORTEM TISSUE" (2017). Theses and Dissertations--Chemistry. 86. https://uknowledge.uky.edu/chemistry_etds/86 This Doctoral Dissertation is brought to you for free and open access by the Chemistry at UKnowledge. It has been accepted for inclusion in Theses and Dissertations--Chemistry by an authorized administrator of UKnowledge. For more information, please contact [email protected]. STUDENT AGREEMENT: I represent that my thesis or dissertation and abstract are my original work. Proper attribution has been given to all outside sources. I understand that I am solely responsible for obtaining any needed copyright permissions. I have obtained needed written permission statement(s) from the owner(s) of each third-party copyrighted matter to be included in my work, allowing electronic distribution (if such use is not permitted by the fair use doctrine) which will be submitted to UKnowledge as Additional File. I hereby grant to The University of Kentucky and its agents the irrevocable, non-exclusive, and royalty-free license to archive and make accessible my work in whole or in part in all forms of media, now or hereafter known.
    [Show full text]
  • Burrowing Time of the Three Indonesian Hippoid Crabs After Artificial Dislodgment
    ILMU KELAUTAN September 2016 Vol 21(3):135-142 ISSN 0853-7291 Burrowing Time of the Three Indonesian Hippoid Crabs After Artificial Dislodgment Yusli Wardiatno*, Yuyun Qonita, and Agus Alim Hakim Department of Aquatic Resources Management, Faculty of Fisheries and Marine Sciences Bogor Agricultural University, Kampus IPB Darmaga, Bogor, Indonesia 16680 Email: [email protected] Abstract Three species of hippoid crabs are the target species of intertidal fishery along coastal line in District Cilacap, south Java; namely Emerita emeritus, Hippa adactyla and Albunea symmista. In Adipala sandy beach, Cilacap an experiment was conducted to reveal the burrowing time and velocity of the crabs. The experiment was performed by removing the crabs from their burrows, measuring their carapace length, and releasing them immediately on the substrate. Burrowing time was measured from the start of burrowing to the disappearance of the entire carapace under the sediment surface. Among the three species, E. emeritus had the fastest burrowing time. As a consequence in terms of velocity, the burrowing velocity of Albunea symmista was higher than that of Hippa adactyla and Emerita emeritus; meaning that with the same size A. symmista needs longer time to burrow. By evaluating with other previous studies, the burrowing time and burrowing velocity of the three sand crabs were comparable. The ability of fast burrowing in the three species seems likely to be the advantage for their survival in large wave disturbed coarse sandy habitat and for their ability to widely exist along the sandy coast of south Java. Keywords: behavior; Indian ocean; intertidal; sand crab; south Java; swash zone Introduction destination beaches of south Java.
    [Show full text]
  • Studio Della Modulazione Ormonale Della Riproduzione E Della Crescita Nei Crostacei Decapodi Mediante L’Uso Di Neurormoni Ricombinanti Ed Anticorpi Specifici
    UNIVERSITÀ DEGLI STUDI DI TRIESTE Sede Amministrativa del Dottorato di Ricerca XX CICLO DEL DOTTORATO DI RICERca IN NEUROSCIENZE Studio della modulazione ormonale della riproduzione e della crescita nei Crostacei Decapodi mediante l’uso di neurormoni ricombinanti ed anticorpi specifici (Settore scientifico-disciplinare BIO/05) DOTTORANDO COORDINATORE DEL COLLEGIO DEI ALESSANDRO MOSCO DOCENTI Prof.sa PAOLA LORENZON Università degli Studi di Trieste TUTORE Dott. PIERO G. GIULIANINI Università degli Studi di Trieste CO-TUTORE Dott. PAOLO EDOMI Università degli Studi di Trieste Anno Accademico 2006-2007 Indice Introduzione 1 Localizzazione del cHH 2 Struttura del cHH 4 Maturazione 9 Funzione 10 Manipolazione ormonale 12 Scopo della tesi 13 Materiali e metodi 14 Clonaggio del cDNA nel vettore di espressione 14 Clonaggio del Asl-rcHH-Gly-Lys 14 Clonaggio del Asl-rcHH-Gly 14 Espressione dei peptidi ricombinanti 14 Refolding e purificazione 16 Reazione di ammidazione 16 Spettrometria di massa 16 Dicroismo circolare 17 Estratto di ghiandole del seno 17 Purificazione del cHH nativo 17 Misura della concentrazione 17 Produzione e purificazione di anticorpi anti-Asl-rcHH-Val-Gly 18 ELISA 18 Stabulazione degli animali 19 Saggi biologici in vivo 19 Inibizione del cHH in A. leptodactylus 20 Risultati 21 Clonaggio del cDNA codificante per cHH-Gly-Lys e cHH-Gly 21 Espressione dei peptidi ricombinanti 21 Refolding e purificazione 21 Ammidazione 22 Caratterizzazione biochimica 22 Quantificazione del cHH 25 Saggi omologhi 26 Saggi eterologhi 29 Risposta iperglicemica
    [Show full text]
  • Invertebrate ID Guide
    11/13/13 1 This book is a compilation of identification resources for invertebrates found in stomach samples. By no means is it a complete list of all possible prey types. It is simply what has been found in past ChesMMAP and NEAMAP diet studies. A copy of this document is stored in both the ChesMMAP and NEAMAP lab network drives in a folder called ID Guides, along with other useful identification keys, articles, documents, and photos. If you want to see a larger version of any of the images in this document you can simply open the file and zoom in on the picture, or you can open the original file for the photo by navigating to the appropriate subfolder within the Fisheries Gut Lab folder. Other useful links for identification: Isopods http://www.19thcenturyscience.org/HMSC/HMSC-Reports/Zool-33/htm/doc.html http://www.19thcenturyscience.org/HMSC/HMSC-Reports/Zool-48/htm/doc.html Polychaetes http://web.vims.edu/bio/benthic/polychaete.html http://www.19thcenturyscience.org/HMSC/HMSC-Reports/Zool-34/htm/doc.html Cephalopods http://www.19thcenturyscience.org/HMSC/HMSC-Reports/Zool-44/htm/doc.html Amphipods http://www.19thcenturyscience.org/HMSC/HMSC-Reports/Zool-67/htm/doc.html Molluscs http://www.oceanica.cofc.edu/shellguide/ http://www.jaxshells.org/slife4.htm Bivalves http://www.jaxshells.org/atlanticb.htm Gastropods http://www.jaxshells.org/atlantic.htm Crustaceans http://www.jaxshells.org/slifex26.htm Echinoderms http://www.jaxshells.org/eich26.htm 2 PROTOZOA (FORAMINIFERA) ................................................................................................................................ 4 PORIFERA (SPONGES) ............................................................................................................................................... 4 CNIDARIA (JELLYFISHES, HYDROIDS, SEA ANEMONES) ............................................................................... 4 CTENOPHORA (COMB JELLIES)............................................................................................................................
    [Show full text]
  • In the United States District Court for the Northern District of Ohio Western Division
    Case: 3:16-cv-02724-JRK Doc #: 19 Filed: 01/29/18 1 of 19. PageID #: <pageID> IN THE UNITED STATES DISTRICT COURT FOR THE NORTHERN DISTRICT OF OHIO WESTERN DIVISION KAYLA D. SPENCER, Case No. 3:16 CV 2724 Plaintiff, v. Magistrate Judge James R. Knepp, II COMMISSIONER OF SOCIAL SECURITY, Defendant. MEMORANDUM OPINION AND ORDER INTRODUCTION Plaintiff Kayla D. Spencer (“Plaintiff”) filed a Complaint against the Commissioner of Social Security (“Commissioner”) seeking judicial review of the Commissioner’s decision to deny disability insurance benefits (“DIB”) and supplemental security income (“SSI”). (Doc. 1). The district court has jurisdiction under 42 U.S.C. §§ 1383(c) and 405(g). The parties consented to the undersigned’s exercise of jurisdiction in accordance with 28 U.S.C. § 636(c) and Civil Rule 73. (Non-document entry dated February 13, 2017). For the reasons stated below, the undersigned affirms the decision of the Commissioner. PROCEDURAL BACKGROUND Plaintiff filed for DIB and SSI in October 2011, alleging a disability onset date of September 30, 2010. (Tr. 326-35). Her claims were denied initially and upon reconsideration. (Tr. 198-203, 208-12). Plaintiff then requested a hearing before an administrative law judge (“ALJ”). (Tr. 213-14). Plaintiff (represented by counsel), and a vocational expert (“VE”) testified at a hearing before the ALJ on August 21, 2013. (Tr. 76-120). On October 18, 2013, the ALJ found Plaintiff not disabled in a written decision. (Tr. 169-92). On April 6, 2015, the Appeals Council Case: 3:16-cv-02724-JRK Doc #: 19 Filed: 01/29/18 2 of 19.
    [Show full text]
  • Sex-Specific Growth and Survival in the Mole Crab Emerita Portoricensis (Schmitt)
    0045258 JOURNALOF CRUSTACEANBIOLOGY, 11(1): 103-112, 1991 SEX-SPECIFIC GROWTH AND SURVIVAL IN THE MOLE CRAB EMERITA PORTORICENSIS (SCHMITT) Miguel P. Sastre ABSTRACT The anomurancrab Emerita portoricensis often occursin the intertidalzone of sandybeaches in the Caribbean.Four sites on PuertoRico, within Mayagiiezand Aiiasco bays, were sampled for 2 yearsin orderto constructsize-frequency distributions and determinerelative growth and survival of each sex. Emeritaportoricensis was regardedas a dioecious species, since no inter- mediateforms were found in any of the 30,755 specimens.Size frequencydistributions of male and female mole crabs were not significantlydifferent between sites. However, the maximum length of the carapaceof male mole crabs (11 mm) was shorterthan that of females (19 mm). Growthcurves based on size-frequencydistributions indicated that females grew significantly fasterthan males. Densities of males were greatestfollowing recruitment. The percentagesur- vival of youngindividuals (< 5 months of age)was higherfor males than femalesand percentage survival of older mole crabs was greaterfor females. Results of this study indicate that sexual heterogeneitiesin size structureof populations can be explained by differentialgrowth and survival of the sexes. Mole crabs of the genus Emerita (Ano- spermatophoresto pleopods of larger fe- mura: Hippidae) are commonly encoun- males (MacGintie, 1938; Wharton, 1942; tered in the intertidalzone of temperateand Knox and Boolotian, 1963; Efford, 1967; tropical sandy beaches. These crabs usually Diaz, 1980; Subramoniam, 1984). aggregateand migrate alongshore (Dillery Caribbean mole crabs, E. portoricensis and Knapp, 1970). Smaller crabs typically (Schmitt, 1935), spawn continuously occur in the upper portion of the littoral throughout the year. The proportion of zone, while largerorganisms are usually lo- ovigerous females increases with growth to cated at lower levels within this zone (Ef- larger size classes.
    [Show full text]