JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing:
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| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in engl. und dt. Sprache |
| Beschreibung: | XV, 99 Bl. Ill., graph. Darst. |
Internformat
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Datensatz im Suchindex
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| adam_text | IMAGE 1
TABLE OF CONTENTS
SUMMARY V
ZUSAMMENFASSUNG VI
ACKNOWLEDGEMENTS VIII
ABBREVIATIONS IX
LIST OF FIGURES XIV
1 INTRODUCTION 1
1.1 THE SKIN 1
1.1.1 THE DERMIS 2
1.1.2 THE BASEMENT MEMBRANE 2
1.1.3 THE EPIDERMIS 3
1.2 CUTANEOUS WOUND HEALING 5
1.2.1 THE FIBRIN CLOT 5
1.2.2 INFLAMMATION 6
1.2.3 NEW TISSUE FORMATION 6
1.2.4 TISSUE REMODELLING AND SCARING 7
1.3 KERATINOCYTE-FIBROBLAST COMMUNICATION IN SKIN 8
1.3.1 IMPACT OF FIBROBLASTS ON KERATINOCYTE BEHAVIOUR 8
1.3.2 GROWTH FACTORS AND CYTOKINES DURING WOUND HEALING 9
1.3.3 IN VITRO STUDIES OF FIBROBLAST-KERATINOCYTE CROSSTALK 11
1.3.4 ROLE OF AP-1 IN THE DOUBLE PARACRINE INTERACTIONS BETWEEN
KERATINOCYTES
AND FIBROBLASTS 11
1.4 THE C-JUN N-TERMINAL KINASES 13
1.4.1 THE STRUCTURE OF JNK PROTEINS 13
1.4.2 THE JNK SIGNALLING CASCADE 14
1.4.3 THE FUNCTIONS OF JNK1 OR JNK2 IN VIVO 15
1.4.4 JNK1 AND JNK2 IN EPIDERMAL MORPHOGENESIS AND WOUND HEALING 16
1.5 AIMS 18
2 MATERIAL 19
2.1 EQUIPMENT AND CONSUMABLES 19
HTTP://D-NB.INFO/1035537516
IMAGE 2
2.2 CHEMICALS 20
2.3 ANTIBODIES 22
2.3.1 PRIMARY ANTIBODIES 22
2.3.2 SECONDARY ANTIBODIES 22
2.4 OLIGONUCLEOTIDES 22
2.5 ENZYMES AND MOLECULAR BIOLOGY REAGENTS 23
2.6 KITS 24
2.7 INHIBITORS 24
2.8 MOLECULAR WEIGHT MARKERS 24
2.9 CELL CULTURE 24
2.9.1 CELL CULTURE MEDIA AND REAGENTS 24
2.9.2 CELLS AND CELL LINES 26
2.10 BUFFERS AND SOLUTIONS 26
3 METHODS 28
3.1 MOLECULAR BIOLOGY METHODS 28
3.1.1 ISOLATION OF TOTAL RNA USING TRIZOL 28
3.1.2 DNASE DIGESTION OF RNA 28
3.1.3 ISOLATION OF TOTAL RNA USING RNEASY MINI KIT 29
3.1.4 REVERSE TRANSCRIPTION 29
3.1.5 AMPLIFICATION OF DNA BY POLYMERASE CHAIN REACTION (PCR) 29
3.1.6 QUANTITATIVE REAL-TIME PCR (QRT-PCR) 30
3.2 PROTEIN BIOCHEMISTRY METHODS 31
3.2.1 PREPARATION OF PROTEIN EXTRACTS 31
3.2.2 DETERMINATION OF PROTEIN CONCENTRATION ACCORDING TO LOWRY 31
3.2.3 SDS POLYACRYLAMIDE GEL ELECTROPHORESIS (SDS-PAGE) 31
3.2.4 TRANSFER OF PROTEINS TO NITROCELLULOSE MEMBRANES (WESTERN BLOT) 32
3.3 HISTOLOGY METHODS 33
3.3.1 CO-LMMUNOFLUORESCENCE OF K10 AND LORICRIN 33
3.3.2 IMMUNOFLUORESCENT DETECTION OF BRDU 33
3.3.3 IMAGE ACQUISITION 34
IMAGE 3
III
3.4 GLOBAL GENE EXPRESSION ANALYSIS 34
3.4.1 ISOLATION OF TOTAL RNA 34
3.4.2 SUPPLY OF SERVICE BY THE GENOMICS AND PROTEOMICS CORE FACILITY
(DKFZ) 34
3.4.3 PROCESSING OF MICROARRAY DATA 34
3.5 CELL CULTURE 35
3.5.1 CALCIUM DEPLETION FROM FETAL BOVINE SERUM 35
3.5.2 ISOLATION OF PRIMARY KERATINOCYTES FROM HUMAN FORESKIN 36
3.5.3 CULTIVATION AND PASSAGING OF MOUSE EMBRYONIC FIBROBLASTS 37
3.5.4 FREEZING AND THAWING OF CELLS 38
3.5.5 DOSE RESPONSE CURVE 38
3.5.6 CO-CULTIVATION OF FIBROBLASTS AND KERATINOCYTES IN A TRANSWELL
SYSTEM 38
3.5.7 CELL PROLIFERATION ASSAY 39
4 RESULTS 40
4.1 IN VITRO CO-CULTURE CONDITIONS TO STUDY KERATINOCYTE-FIBROBLAST
INTERACTIONS
DURING RE-EPITHELIALIZATION IN CUTANEOUS WOUND HEALING 40
4.1.1 DETERMINATION OF THE APPROPRIATE CACI 2 CONCENTRATION IN
CO-CULTURE MEDIUM 40
4.1.2 THE MODIFIED CO-CULTURE SYSTEM 4 1
4.1.3 FIBROBLASTS PROMOTE KERATINOCYTE BEHAVIOUR WHEN TRIGGERED FOR
DIFFERENTIATION . 43
4.2 KERATINOCYTE PROLIFERATION AND TERMINAL DIFFERENTIATION UPON LOSS OF
DERMAL JNK 46
4.2.1 KERATINOCYTE PROLIFERATION IN CO-CULTURES WITH WT OR JNK-DEFICIENT
FIBROBLASTS 46
4.2.2 KERATINOCYTE DIFFERENTIATION DEPENDS ON DERMAL JNK 50
4.2.3 BOTH, DERMAL JNK1 AND JNK2 PROMOTE KERATINOCYTE TERMINAL
DIFFERENTIATION 57
4.3 IDENTIFICATION OF JNK-DEPENDENT SOLUBLE FACTOR(S) 61
4.3.1 KERATINOCYTE TERMINAL DIFFERENTIATION IS DEPENDENT ON CONSTANT
PRESENCE OF
DERMAL JNK-DEPENDENT SOLUBLE FACTOR(S) 61
4.3.2 JNK-DEPENDENT GENE EXPRESSION IN FIBROBLASTS ENCODING SOLUBLE
FACTORS WITH
POTENTIAL IMPACT ON KERATINOCYTE DIFFERENTIATION 70
5 DISCUSSION 73
5.1 IMPROVED CO-CULTURE CONDITIONS AS A PREREQUISITE TO ANALYZE THE
IMPACT OF
FIBROBLASTS ON KERATINOCYTE DIFFERENTIATION 73
5.2 DERMAL JNK1 AND JNK2 PROMOTE KERATINOCYTE DIFFERENTIATION DURING
RE-EPITHELIALISATION 75
IMAGE 4
5.3 CONTRIBUTION OF KERATINOCYTE-DERIVED STIMULI TO THE DOUBLE PARACRINE
CROSSTALK
WITH FIBROBLASTS 76
5.3.1 KERATINOCYTE-DERIVED STIMULI CONTROLLING JNK ACTIVITY IN
FIBROBLASTS 76
5.3.2 KERATINOCYTES REGULATE FIBROBLAST GENE EXPRESSION 78
5.4 ROLE OF JNK SUBSTRATES IN THE REGULATION OF JNK-DEPENDENT ACTIVITY
ON
KERATINOCYTE DIFFERENTIATION 78
5.5 IDENTIFICATION OF DERMAL JNK-REGULATED FACTOR(S) PROMOTING
KERATINOCYTE
DIFFERENTIATION 80
5.6 CONCLUSION AND PERSPECTIVES 85
6 REFERENCES 86
CONFIRMATION 100
|
| any_adam_object | 1 |
| author | Schumacher, Marion |
| author_GND | (DE-588)1033023027 |
| author_facet | Schumacher, Marion |
| author_role | aut |
| author_sort | Schumacher, Marion |
| author_variant | m s ms |
| building | Verbundindex |
| bvnumber | BV040936249 |
| ctrlnum | (OCoLC)853280986 (DE-599)BSZ380504170 |
| dewey-full | 570 610 |
| dewey-hundreds | 500 - Natural sciences and mathematics 600 - Technology (Applied sciences) |
| dewey-ones | 570 - Biology 610 - Medicine and health |
| dewey-raw | 570 610 |
| dewey-search | 570 610 |
| dewey-sort | 3570 |
| dewey-tens | 570 - Biology 610 - Medicine and health |
| discipline | Biologie Medizin |
| format | Thesis Book |
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| indexdate | 2024-07-10T00:35:44Z |
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| language | English |
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| physical | XV, 99 Bl. Ill., graph. Darst. |
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| spelling | Schumacher, Marion Verfasser (DE-588)1033023027 aut JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing [presented by Marion Schumacher] 2013 XV, 99 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in engl. und dt. Sprache Heidelberg, Univ., Diss., 2013 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025915106&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schumacher, Marion JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing |
| subject_GND | (DE-588)4113937-9 |
| title | JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing |
| title_auth | JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing |
| title_exact_search | JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing |
| title_full | JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing [presented by Marion Schumacher] |
| title_fullStr | JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing [presented by Marion Schumacher] |
| title_full_unstemmed | JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing [presented by Marion Schumacher] |
| title_short | JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing |
| title_sort | jnk dependent dermal genetic program controls interdependent keratinocyte fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025915106&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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