Holdings: Extracellular matrix-cell interactions regulating zebrafish fin development

Extracellular matrix-cell interactions regulating zebrafish fin development:

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Main Author:	Konrad Richetti, Stefania 1986- (Author)
Format:	Thesis Microfilm Book
Language:	English
Published:	Köln 2015
Subjects:	Hochschulschrift
Online Access:	Inhaltsverzeichnis Inhaltsverzeichnis
Item Description:	Zusammenfassung in deutscher und englischer Sprache
Physical Description:	2 Mikrofiches (150 Seiten) Illustrationen, Diagramme

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adam_text	Titel: Extracellular matrix-cell interactions regulating zebrafish fin development Autor: Konrad Richetti, Stefania Jahr: 2015 Table of Contents Acknowledgements.................................................................................................................................3 Summary.................................................................................................................................................7 Zusammenfassung...................................9 1 Introduction...............................11 1.1 Teleost and mammalian epidermis: similarities and differences.........................................12 1.1.1 Mammalian epidermis development............................................................................12 1.1.2 Zebrafish epidermis development................................................................................12 1.1.3 Regulation of epidermal differentiation in fish and mammals.....................................16 1.2 Basement membrane............................................................................................................16 1.2.1 Laminins.............................................18 1.2.2 The Fibulins...................................................................................................................19 1.2.3 Nidogens.......................................................................................................................24 1.3 Zebrafish as a model to study the BM..................................................................................27 1.4 Zebrafish mutants with defects in fin development.............................................................28 1.5 Initial teleost fin development..............................................................................................90 1.5.1 Establishment of a fin fold............................................................................................90 1.5.2 Fin mesenchymal cell migration...................................................................................33 1.6 Objectives..............................................................................................................................36 1.6.1 Control of fin mesenchymal cell migration during fin development............................36 1.6.2 Role of epidermal-dermal junction in fin development...............................................36 2 Material and Methods..................................................................................................................37 2.1 Molecular Biology.................................................................................................................37 2.2 Isolation of genomic DNA.....................................................................................................37 2.3 Genotyping............................................................................................................................37 2.4 Microinjection.......................................................................................................................38 2.5 Morpholinos..........................................................................................................................38 2.6 Embryo fixation.....................................................................................................................39 2.7 In situ probe synthesis..........................................................................................................39 2.8 Whole-mount in situ hybridization.......................................................................................40 2.9 Durcupan sectioning.............................................................................................................41 2.10 Immunohistochemistry.........................................................................................................42 2.11 Imaging.......................................43 2.12 Image! programming for analysis of E-Cad staining.............................................................44 2.13 Western Blots........................................................................................................................45 2.14 Drug treatments....................................................................................................................46 2.15 Animal care...........................................................................................................................46 2.16 Transmission Electron Microscopy.......................................................................................47 2.16.1 Embryo fixation.............................................................................................................47 2.16.2 Epoxy Resin Embedding and Sectioning.......................................................................47 Control of fin mesenchymal cell migration during fin development............................................49 3 Results...........................................................................................................................................50 3.1 Fin mesenchymal cell migration...........................................................................................50 3.2 Expression of fibronectin during fin development...............................................................52 3.3 Fibronectin accumulates at somite borders and neural tube during zebrafish development......................................................................................................................................54 3.4 Expression of integrins during zebrafish development........................................................56 3.5 Mutation infibronectinlb does not affect FMC migration...................................................63 3.6 Inhibition of interaction between fin mesenchymal cells and the extracellular matrix results in defects in fin mesenchymal cell morphology...............................................................................65 4 Discussion......................................................................................................................................68 4.1 Zebrafish fin subepidermal space is invaded by fin mesenchymal cells at 26 hours post- fertilization ........................................................................................................................................68 4.2 Switch between RGD-binding Integrins as a trigger for migration.......................................69 4.3 ltgavP3 mediated apical migration: a role of Flemicentin-2?...............................................73 4.4 Interaction with the Fibronectin matrix is necessary for proper migration.........................75 4.5 Interaction with actinotrichia via Itga2\|31 is needed for orientation of protrusions...........77 4.6 Feeling and shaping the matrix: a model for fin mesenchymal cell migration.....................78 5 Results...........................................................................................................................................81 5.1 Nidogens in zebrafish development.....................................................................................81 5.1.1 nidogen la....................................................................................................................81 5.1.2 nidogen lb....................................................................................................................82 5.1.3 nidogen 2a....................................................................................................................84 5.2 Nidogen 2a protein localization............................................................................................85 5.3 nidogen 2a knockdown.........................................................................................................86 5.4 Efficient nidogen2a knockdown............................................................................................88 5.5 Knockdown of nidogen lb....................................................................................................90 5.6 Synergistic interaction between nidogen lb and nidogen 2a..............................................92 5.7 Mouse Nidogen2 mRNA rescues fin fold reduction phenotype...........................................94 5.8 Synergistic interaction between laminin aS and nidogen 2a...............................................95 5.9 Synergistic interaction between hemicentin 1 and nidogen 2a............................................97 5.10 Loss of Hemicentin 1 and Laminin ct5 does not affect Nidogen 2a protein distribution.... 102 5.11 Laminin clusters are observed in the apical portion of the fin fold in nidogen 2a morphants and nidogen 2a/hemicentinl double morphants...........................................................................104 5.12 Knockdown of nidogen2a results in reduced adhesion between EBL................................106 ........................................................................................................................................................109 5.13 Knockdown of nid2a affects E-cadherin localization..........................................................110 5.14 Distribution of E-cad in the cell membrane is affected by loss of nid2a............................Ill 5.15 Cell size is affected by loss of nidogen 2a, nidogen lb and hemicentin 1..........................113 5.16 Simultaneous knockdown of nidogen 2a and hemicentin 1 result in a high variability of cell shapes..........................................................................................................................................................115 5.17 Impaired interaction between Nidogens and Hemicentin -1 affects the organization of the fin ECM and adhesion between basal keratinocytes......................................................................117 6 Discussion....................................................................................................................................122 6.1 Nidogen 2a as a regulator of fin epidermal integrity..........................................................123 6.2 Nidogen 2a and Nidogen lb - partial redundant roles in fin development.......................126 6.3 Nidogen 2a and Laminin a5 function in the same pathway controlling fin development. 127 6.4 Interaction between Nidogen 2a and Hemicentin 1 is needed for epidermal-dermal junction maintenance.....................................................................................................................127 7 References..................................................................................................................................130 8 Appendix.....................................................................................................................................142 8.1 List of figures.......................................................................................................................142 8.2 List of tables........................................................................................................................143 Erklarung.............................................................................................................................................147 Lebenslauf...........................................................................................................................................148
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author	Konrad Richetti, Stefania 1986-
author_GND	(DE-588)1098455738
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dewey-full	573.7938617482
dewey-hundreds	500 - Natural sciences and mathematics
dewey-ones	573 - Specific physiological systems in animals
dewey-raw	573.7938617482
dewey-search	573.7938617482
dewey-sort	3573.7938617482
dewey-tens	570 - Biology
discipline	Biologie
format	Thesis Microfilm Book
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spelling	Konrad Richetti, Stefania 1986- Verfasser (DE-588)1098455738 aut Extracellular matrix-cell interactions regulating zebrafish fin development vorgelegt von Stefania Konrad Richetti Köln 2015 2 Mikrofiches (150 Seiten) Illustrationen, Diagramme txt rdacontent h rdamedia he rdacarrier 24x Zusammenfassung in deutscher und englischer Sprache Dissertation Universität zu Köln 2015 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1103249525/04 Inhaltsverzeichnis HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029230425&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis
spellingShingle	Konrad Richetti, Stefania 1986- Extracellular matrix-cell interactions regulating zebrafish fin development
subject_GND	(DE-588)4113937-9
title	Extracellular matrix-cell interactions regulating zebrafish fin development
title_auth	Extracellular matrix-cell interactions regulating zebrafish fin development
title_exact_search	Extracellular matrix-cell interactions regulating zebrafish fin development
title_full	Extracellular matrix-cell interactions regulating zebrafish fin development vorgelegt von Stefania Konrad Richetti
title_fullStr	Extracellular matrix-cell interactions regulating zebrafish fin development vorgelegt von Stefania Konrad Richetti
title_full_unstemmed	Extracellular matrix-cell interactions regulating zebrafish fin development vorgelegt von Stefania Konrad Richetti
title_short	Extracellular matrix-cell interactions regulating zebrafish fin development
title_sort	extracellular matrix cell interactions regulating zebrafish fin development
topic_facet	Hochschulschrift
url	http://d-nb.info/1103249525/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029230425&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
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