Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish:
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2012
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Zsfassung in dt. und engl. Sprache |
| Physical Description: | 132 S. Ill., graph. Darst. |
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| 245 | 1 | 0 | |a Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish |c vorgelegt von Reza Thomas Ramezani |
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| adam_text | Titel: Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adu
Autor: Ramezani, Reza Thomas
Jahr: 2012
Table of Contents
Table of Contents.........................................................................................3
Summary......................................................................................................6
/ Contribution ofextracellular matrix to embryonic fin development.........................6
II Ciitaneous wound closure in adult zebraflsh............................................................8
Zusammenfassung.....................................................................................10
/ Beteiligung der extrazelluldren Matrix an embryonaler Flossenenrwicklung........10
II Kutaner Wundverschluss im adulien Zebraftsch....................................................12
Contribution ofextracellular matrix to embryonic fin development... 14
I Introduction..............................................................................................................15
1.1 The dermal-epidermal basement membrane.....................................................15
1.2 Nidogens............................................................................................................18
1.3 Embryonic fin development as a model for BM assembly...............................21
1.3.1 Embryonic fin fold formation...............................................................................21
1.3.2 Mutations affecting fin fold morphogenesis.........................................................23
1.4 Aims of the project............................................................................................25
UResuIts.....................................................................................................................26
II. I Structure and évolution of nidogen proteins....................................................26
11.2 Zebraflsh nidogen la.......................................................................................28
11.2.1 Expression ofnidla during embryogenesis........................................................28
II. 2.2 5 RACE and morpholino (MO) knockdown ofnidla..........................................29
11.3 Zebrafish nidogen 1b.......................................................................................30
11.4 Zebraflsh nidogen 2b.......................................................................................32
11.5 Zebrafish nidogen 2a.......................................................................................33
11.5.1 Developmenlal expression ofnid2a....................................................................33
11.5.2 Protein localisation ofNid2a..............................................................................35
11.6 Contribution ot nid2a to embryonic fin development.....................................36
11.6.1 MO knockdown of nid2a gène actixity................................................................36
11.6.2 Genetic interaction ofnid2a andhmcnl.............................................................38
II. 6.3 Comparative time-series ofnid2a morphants wilhfra and ne!...........................40
11. 6.4 Comparison of morphant and mutant BM composition......................................41
11.6.5 Transmission électron microscopy (TEM)..........................................................43
11.7 Characterisation of a nevv tin blister mutant....................................................47
11.7.1 Phenolype of the miles apart-Iike (2.4102) mutant..............................................47
11.7.2 Genetic mapping ofmil-like (2Al02)..................................................................49
11.7.3 Expression of slpr2 andspns2...........................................................................50
11.7.4 BM constitution in mil mutants...........................................................................51
11.7.5 Fibroneclin in mil and ton..................................................................................53
11.7.6 Hemicentin 2 and Collagen 11 in mil and toh......................................................55
III Discussion.............................................................................................................57
111.1 The rôle of nidogens in embryonic fin development......................................57
111.2 New insights into ECM assembly during fin fold formation.........................61
111.3 Amodel for the maintenance of fin fold formation........................................64
Cutaneous wound closure in adult zebrafish..........................................66
I Introduction..............................................................................................................67
1.1 Adult zebrafish skin..........................................................................................67
1.2 Cutaneous wound closure..................................................................................70
1.3 Mutagenesis screening in zebraflsh...................................................................72
1.4 Aimsof the project............................................................................................74
II Results.....................................................................................................................75
11.1 Adult wound closure screen.............................................................................75
11.2 Live imaging of closing wounds......................................................................78
11.3 Scanning électron microscopy (SEM) of adult wound closure.......................80
11.3.1 Architecture of the superficial cell layer.............................................................80
11.3.2 Architecture ofwounded skin..............................................................................81
11.3.3 Cell shape changes during wound closure..........................................................83
II. 3.4 Examination ofthe leading edge cells................................................................85
11.4 Génération of transgenic lines.........................................................................87
III Discussion.............................................................................................................90
111.1 The fast wound closure in adult zebrafish......................................................90
111.2 Models for cutaneous wound closure in fish..................................................92
111.3 A working-model for adult zebrafish wound closure.....................................95
111.4 Conclusions and outlook, the need for transgenic lines.................................97
Materials and Methods...........................................................................100
I Expérimental procédures.......................................................................................101
1.1 Zebrafish strains and maintenance..................................................................101
1.2 Genetic mapping............................................... 101
1.3 5 RACE and cDNA synthesis.........................................................................101
1.4 Microinjection.................................................. ...102
1.4.1 Morpholinos.................................................... ]02
1.4.2 MO control......................................... 102
1.4.3 mRNA synthesis.............................................. 102
1.5 Semiquantitative RT-PCR...............................................................................103
1.6 Tissue fixation.................................................................................................103
1.7 Whole-mount in situ hybridization.................................................................103
1.7.1 In Situ probe synthesis.......................................................................................104
1.8 Immunohistochemistry....................................................................................104
1.9 Microscopy......................................................................................................104
1.9.1 Scanning électron microscopy...........................................................................105
1.9.2 Transmission électron microscopy.....................................................................105
1.9.3 Live imaging......................................................................................................105
1.10 Durcupan embedding and sectioning............................................................105
1.11 Wound closure assay and screen...................................................................106
1.12 Gateway-cloning of transgenic constructs....................................................106
II Materials...............................................................................................................107
Bibliography.............................................................................................110
Abbreviations...........................................................................................122
Appendix..................................................................................................124
I List of Figures........................................................................................................125
II List of Tables........................................................................................................127
III Table ofscreened F3 familles.............................................................................128
II keratin 4 (krl4)promoter séquence.....................................................................133
Danksagung..............................................................................................134
Erklârung.................................................................................................135
Curriculum vitae.....................................................................................136
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| language | English |
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| spelling | Ramezani, Reza Thomas 1982- Verfasser (DE-588)1023546868 aut Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish vorgelegt von Reza Thomas Ramezani 2012 132 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. und engl. Sprache Köln, Univ., Diss., 2012 (DE-588)4113937-9 Hochschulschrift gnd-content HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025171943&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Ramezani, Reza Thomas 1982- Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish |
| subject_GND | (DE-588)4113937-9 |
| title | Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish |
| title_auth | Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish |
| title_exact_search | Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish |
| title_full | Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish vorgelegt von Reza Thomas Ramezani |
| title_fullStr | Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish vorgelegt von Reza Thomas Ramezani |
| title_full_unstemmed | Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish vorgelegt von Reza Thomas Ramezani |
| title_short | Contribution of extracellular matrix to embryonic fin development and Cutaneous wound closure in adult zebrafish |
| title_sort | contribution of extracellular matrix to embryonic fin development and cutaneous wound closure in adult zebrafish |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025171943&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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