The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts:
Saved in:
| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2013
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | XII, 164 S. Ill., graph. Darst. |
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MARC
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| 502 | |a Münster (Westfalen), Univ., Diss., 2013 | ||
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Record in the Search Index
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|---|---|
| adam_text | IMAGE 1
CONTENTS
1. INTRODUCTION 1
1.1. ENDOTHELIAL BARRIER FUNCTION 3
1.2. ORGANIZATION OF ENDOTHELIAL CELL-CELL CONTACTS 4
1.2.1. TIGHT JUNCTIONS 5
1.2.2. ADHERENS JUNCTION 9
1.2.3. COMPLEXUS ADHAERENTES 19
1.2.4. GAP JUNCTIONS 19
1.2.5. ADHESIVE PROTEINS OF THE INTERENDOTHELIAL CLEFT 19
1.3. LEUKOCYTE ADHESION CASCADE 20
1.3.1. CAPTURING, ROLLING AND SLOW ROLLING 22
1.3.2. LEUKOCYTE ACTIVATION AND ARREST 22
1.3.3. FIRM ADHESION AND CRAWLING 23
1.3.4. TRANSENDOTHELIAL CELL MIGRATION 24
1.4. ROLE OF VE-CADHERIN TYROSINE PHOSPHORYLATION FOR ENDOTHELIAL CELL
CONTACTS . . . . . . 26
1.4.1. FACTORS AND MECHANISMS T H A T ALTER VE-CADHERINS PHOSPHORYLATION
29
1.4.2. VE-CADHERIN PHOSPHORYLATION AND INHIBITION OF CELL PROLIFERATION
35
1.4.3. LEUKOCYTES AFFECTING VE-CADHERIN PHOSPHORYLATION 35
2. METHODS AND MATERIALS 3 9
2.1. METHODS 39
2.1.1. BIOCHEMISTRY 39
2.1.2. MOLECULAR BIOLOGY 43
2.1.3. CELL BIOLOGY 50
2.1.4. IN VIVO 56
VII
HTTP://D-NB.INFO/1032978775
IMAGE 2
2.2. MATERIALS 62
2.2.1. CHEMICALS AND REAGENTS 62
2.2.2. KITS 65
2.2.3. ANTIBODIES 66
2.2.4. ENZYMES 67
2.2.5. ISOTOPES 67
2.2.6. BUFFERS AND MEDIA 68
2.2.7. CELL LINES 73
2.2.8. MOUSE STRAINS 73
2.2.9. CONSUMABLES 74
2.2.10. EQUIPMENT 75
2.2.11. SOFTWARE 78
3. RESULTS 7 9
3.1. IN VITRO TRANSMIGRATION ACROSS HUVECS EXPRESSING Y / F VE-CADHERIN
MUTANTS . . . . 79
3.1.1. GENERATION OF Y658F, Y731F AND Y658/731F ADENOVIRUSES 79
3.1.2. ADENOVIRAL EXPRESSION OF VE-CADHERIN MUTANTS IN HUVECS 81
3.1.3. OVEREXPRESSION OF MURINE Y / F VE-CADHERIN HAD NO EFFECT ON T E M
83
3.2. GENERATION OF Y658F AND Y731F VE-CADHERIN KNOCK-IN MICE 84
3.2.1. CLONING OF THE Y658F-VEC-RMCE AND Y731F-VEC-RMCE VECTORS 85
3.2.2. GENERATION OF RECOMBINANT ES CELL CLONES USING THE R M C E
TECHNOLOGY . . . . 85
3.2.3. GENERATION OF KNOCK-IN MICE VIA BLASTOCYST INJECTIONS 89
3.2.4. GENERATION OF KNOCK-IN MICE VIA MICROINJECTION OF DNA INTO
PRONUCLEUS . . . 94
3.3. GENERATION OF PHOSPHO SITE-SPECIFIC ANTI VE-CADHERIN ANTIBODIES 94
3.3.1. PREPARATION OF PEPTIDE IMMUNOGENS 96
3.3.2. PRODUCTION OF POLYLCONAL ANTIBODIES IN RABBITS 98
3.3.3. PRODUCTION OF MONOCLONAL ANTIBODIES WITH RATS 104
3.4. PHOSPHORYLATION OF Y685 IS NECESSARY FOR INDUCTION OF VASCULAR
PERMEABILITY I L L
3.4.1. VEGF AND HISTAMINE STIMULATE PHOSPHORYLATION OF Y685 IN HUVECS .
. . . 1 1 1
3.4.2. V E G F AND HISTAMINE-INDUCED VASCULAR PERMEABILITY IS REDUCED IN
SKIN OF
Y685F VE-CADHERIN MICE 115
VIII
IMAGE 3
3.4.3. MICROSPHERES EXTRAVASATE A T SITES OF INDUCED Y685
PHOSPHORYLATION IN MOUSE
CREMASTER 115
3.5. PHOSPHORYLATION OF Y658F AND Y731F IS NECESSARY FOR LEUKOCYTE
TRANSMIGRATION . . . 117
3.5.1. LEUKOCYTES STIMULATE DEPHOSPHORYLATION OF Y731 IN BEND.5 CELLS
117
3.5.2. LEUKOCYTE TRANSMIGRATION IS IMPAIRED IN INFLAMED CREMASTER MUSCLE
OF Y658F
AND Y731F VE-CADHERIN MICE 117
4. DISCUSSION 127
4.1. PRELIMINARY EXPERIMENTS AND GENERATION OF KNOCK-IN MICE AND
ANTIBODIES 128
4.1.1. IN VITRO TRANSENDOTHELIAL MIGRATIONS ASSAYS WITH Y / F
VE-CADHERIN MUTANTS . 128
4.1.2. GENERATION AND CHARACTERIZATION OF Y658F AND Y731F KNOCK-IN MICE
128
4.1.3. GENERATION OF POLYCLONAL AND MONOCLONAL PHOSPHO SITES-SPECIFIC
ANTIBODIES . . 130
4.2. THE TYROSINE RESIDUES OF VE-CADHERIN HAVE DISTINCT ROLES DURING
INFLAMMATION . . . . 131
4.2.1. Y658 AND Y731 ARE NECESSARY FOR EFFICIENT LEUKOCYTE EXTRAVASATION
131
4.2.2. Y685 IS REQUIRED FOR EFFICIENT INDUCTION OF PERMEABILITY 133
A. CURRICULUM VITAE 1 3 5
B. ACKNOWLEDGEMENTS 136
BIBLIOGRAPHY 137
ACRONYMS 1 6 1
IX
|
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| ctrlnum | (OCoLC)844064173 (DE-599)HBZHT017573472 |
| dewey-full | 571.74364355519353 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 571 - Physiology & related subjects |
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| dewey-search | 571.74364355519353 |
| dewey-sort | 3571.74364355519353 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| language | English |
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| physical | XII, 164 S. Ill., graph. Darst. |
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| spelling | Wessel, Florian 1983- Verfasser (DE-588)1032098279 aut The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts vorgelegt von Florian Hubert Erich Wessel 2013 XII, 164 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Münster (Westfalen), Univ., Diss., 2013 Cadherine (DE-588)4314135-3 gnd rswk-swf Endothel (DE-588)4014675-3 gnd rswk-swf Zell-Adhäsionsmolekül (DE-588)4300852-5 gnd rswk-swf Tyrosin (DE-588)4186526-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Endothel (DE-588)4014675-3 s Tyrosin (DE-588)4186526-1 s Cadherine (DE-588)4314135-3 s Zell-Adhäsionsmolekül (DE-588)4300852-5 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025926681&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Wessel, Florian 1983- The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts Cadherine (DE-588)4314135-3 gnd Endothel (DE-588)4014675-3 gnd Zell-Adhäsionsmolekül (DE-588)4300852-5 gnd Tyrosin (DE-588)4186526-1 gnd |
| subject_GND | (DE-588)4314135-3 (DE-588)4014675-3 (DE-588)4300852-5 (DE-588)4186526-1 (DE-588)4113937-9 |
| title | The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts |
| title_auth | The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts |
| title_exact_search | The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts |
| title_full | The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts vorgelegt von Florian Hubert Erich Wessel |
| title_fullStr | The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts vorgelegt von Florian Hubert Erich Wessel |
| title_full_unstemmed | The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts vorgelegt von Florian Hubert Erich Wessel |
| title_short | The role of VE-cadherin tyrosine phosphorylation for the control of endothelial cell contacts |
| title_sort | the role of ve cadherin tyrosine phosphorylation for the control of endothelial cell contacts |
| topic | Cadherine (DE-588)4314135-3 gnd Endothel (DE-588)4014675-3 gnd Zell-Adhäsionsmolekül (DE-588)4300852-5 gnd Tyrosin (DE-588)4186526-1 gnd |
| topic_facet | Cadherine Endothel Zell-Adhäsionsmolekül Tyrosin Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025926681&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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