Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in dt. und engl. Sprache |
| Beschreibung: | 111 S. Ill., graph. Darst. |
Internformat
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| 245 | 1 | 0 | |a Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins |c vorgelegt von Tingting Wen |
| 264 | 1 | |c 2012 | |
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Datensatz im Suchindex
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| adam_text | Titel: Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins
Autor: Wen, Tingting
Jahr: 2012
Table of contents
1. Introduction............................................................................. 1
1.1. Epidermal-dermal junction and epidermolysis bullosa................... 1
1.1.1. Laminin 332........................................................................ 4
1.1.2. Integrins.............................................................................. 12
1.1.2.1. oe6f34 integrin............................................................. 15
1.1.2.2. a2|31 integrin............................................................. 17
1.1.2.3. cc3p1 integrin.............................................................. 17
1.2. Cellular adhesions.......................................................................... 19
1.2.1. Integrins and cell adhesion to the extracellular matrix......... 19
1.2.2. Intercellular junctions........................................................... 21
1.3. Actin cytoskeleton and migration................................................... 22
2. Aim of the study...................................................................... 25
3. Material and methods............................................................. 27
3.1. Material............................................................................................27
3.1.1. Reagents...............................................................................27
3.1.2. Antibodies............................................................................. 28
3.1.2.1. Primary antibodies......................................................28
3.1.2.2. Secondary antibodies.................................................29
3.1.3. Laboratory materials............................................................. 30
3.1.4. Equipment.............................................................................30
3.2. Methods...........................................................................................31
3.2.1. Cell culture............................................................................ 31
3.2.2. Small interference RNA mediated silencing..........................33
3.2.3. Phase contrast microscopy and measurement of cell
proliferation........................................................................... 35
3.2.4. Detection of apoptosis.......................................................... 35
3.2.5. Adhesion assay.....................................................................36
3.2.5.1. Adhesion on different extracellular matrix proteins.... 36
3.2.5.2. Adhesion on plastic.................................................... 36
3.2.6. Time lapse video microscopy................................................37
3.2.6.1. Preparation of cells.....................................................37
3.2.6.2. Equipment, software and recording of migration........37
3.2.7. Protein extraction..................................................................39
3.2.8. SDS-PAGE and immunoblotting...........................................40
3.2.9. Immunofluorescence staining and confocal microscopy...... 41
3.2.10. Statistic analysis................................................................... 43
4. Results..................................................................................... 44
4.1. Silencing of laminin cc3 chain leads to prominent intracellular retention
of laminin fi. chain and increased keratinocyte motility...................44
4.1.1. Efficient down-regulation of laminin oc3 chain upon siRNA
silencing................................................................................44
4.1.2. Knockdown of the laminin a3 chain reduces extracellular
laminin 332 and is associated with an increase of intracellular
laminin 72 chains..................................................................45
4.1.3. Intracellular accumulation of laminin 72 chains does not cause
apoptosis of HaCaT keratinocytes........................................51
4.1.4. Silencing of the laminin a3 chain modifies cell morphology.. 52
4.1.5. Down-regulation of laminin a3 chain leads to enhanced
keratinocyte motility...............................................................54
4.1.6. Down-regulation of laminin a3 chain alters keratinocyte
adhesion ability..................................................................... 56
4.1.7. Down-regulation of laminin a3 chain is associated with up-
regulation of ct2f31 integrin and redistribution of a6(34
integrin.................................................................................. 60
4.1.8. Analysis of actin regulating and signaling proteins............... 64
4.2. Down-regulation of integrin oc2 and a3 subunits results in enhanced
and non-coherent single cell movement......................................... 65
4.2.1. Down-regulation efficiency upon silencing of integrin oc2 and
oc3 subunits............................................................................65
4.2.2. Down-regulation of the integrin oc2 and a3 subunits leads to
morphological alterations......................................................67
4.2.3. Down-regulation of the integrin oc2 and ce3 subunits
enhances non-coherent and single cell migration.................70
4.2.4. Silencing of the integrin ct2 and cc3 subunits activates FAK
and ERK signaling pathway(s)..............................................74
4.2.5. Knockdown of integrin a3 subunit is accompanied by up-
regulation of a2(31 integrin and down-regulation of a6fS4
integrin.................................................................................. 75
4.2.6. Down-regulation of integrin cc3 subunit elevates expression of
fibronectin and laminin 332................................................... 77
5. Discussion................................................................................81
5.1. Role of laminin oc3 chain in regulating the amount of laminin 72 chain
in the cell......................................................................................... 81
5.2. Role of laminin 332 in keratinocyte migration................................. 83
5.3. Roleofa3pi integrin in keratinocytes............................................. 84
5.4. Possible role of a2(51 integrin in contact inhibition in epithelia........ 86
5.5. Role of cxf31 integrins in keratinocyte migration................................87
6. Abbreviations.......................................................................... 90
7. Bibliography............................................................................ 91
Acknowledgements....................................................................109
Eklarung......................................................................................110
Curriculum Vitae.........................................................................111
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| language | English |
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| spelling | Wen, Tingting 1983- Verfasser (DE-588)1033287814 aut Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins vorgelegt von Tingting Wen 2012 111 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=026203451&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Wen, Tingting 1983- Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins |
| subject_GND | (DE-588)4113937-9 |
| title | Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins |
| title_auth | Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins |
| title_exact_search | Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins |
| title_full | Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins vorgelegt von Tingting Wen |
| title_fullStr | Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins vorgelegt von Tingting Wen |
| title_full_unstemmed | Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins vorgelegt von Tingting Wen |
| title_short | Regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins |
| title_sort | regulation of keratinocyte behavior by laminin 332 and alpha beta 1 integrins |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026203451&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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