Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells:
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
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2014
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| Beschreibung: | 124 Bl. Ill., graph. Darst. 30 cm |
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Datensatz im Suchindex
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| adam_text | LYSINE-SPECIFIC DEMETHYLASE 1 REGULATES
DIFFERENTIATION ONSET AND
MIGRATION OF TROPHOBLAST
STEM CELLS
INAUGURALDISSERTATION
ZUR ERLANGUNG DER DOKTORWURDE DER
FAKULTAT FUR BIOLOGIE DER ALBERT-LUDWIGS-UNIVERSITAT
FREIBURG IM BREISGAU
VORGELEGT VON
STEFANIE HOLZ
GEBOREN IN KAISERSLAUTERN
FREIBURG IM BREISGAU, 2014
HTTP://D-NB.INFO/1055852875
CONTENTS
CONTENTS
1 INTRODUCTION 1
1.1 EPIGENETICS 1
1.1.1 CHROMATIN STRUCTURE 2
1.1.2 MODIFICATIONS OF CHROMATIN 3
1.1.3 CHROMATIN READERS 7
1.1.4 CROSS TALK BETWEEN CHROMATIN MODIFICATIONS 8
1.2 HISTONE METHYLATION
AND DEMETHYLATION 9
1.2.1 LYSINE-SPECIFIC DEMETHYLASE 1 (LSDL) 11
1.2.2 LSDL INTERACTING CO-REPRESSOR COMPLEXES 12
1.3 MOUSE PLACENTAL
DEVELOPMENT 13
1.3.1 MOUSE TROPHOBLAST STEM CELL (TSC] LINEAGES 14
1.4 EPIGENETICS DURING
EMBRYONIC DEVELOPMENT 18
1.4.1 EPIGENETIC REGULATION DURING TE LINEAGE-SPECIFICATION 20
1.4.2 FUNCTIONS OF LSD 1 DURING EMBRYONIC DEVELOPMENT 21
2 MATERIAL 23
2.1 CELL LINES
AND CULTURE
MEDIA 23
2.1.1 CELL LINES 23
2.1.2 MEDIA CONTENTS 23
2.1.3 TS CELL CULTURE 23
2.2 BACTERIA 24
2.3 PLASMIDS 24
2.3.1 GENERAL PLASMIDS 24
2.3.2 EXPRESSION PLASMIDS 24
2.4 OLIGONUCLEOTIDES 25
2.4.1 GENOTYPING 25
2.4.2 QRT-PCR 26
2.4.3 MIRNA AND SIRNA 2 7
2.5 ENZYMES 28
2.6 ANTIBODIES 28
2.7 KITS 29
2.8 SOLUTIONS AND
BUFFER 30
2.9 CHEMICALS 32
2.10 DEVICES 33
CONTENTS
3 METHODS 34
3.1 NUCLEIC ACIDS 34
3.1.1 DETERMINATION OF DNA AND RNA CONCENTRATION 34
3.1.2 CLONING WITH THE GATEWAY SYSTEM 34
3.1.3 ANALYTICAL AND QUANTITATIVE PREPARATION OF PLASMID DNA 35
3.1.4 RESTRICTION DIGESTION OF DNA 35
3.1.5 SEPARATION OF DNA FRAGMENTS BY AGAROSE GEL ELECTROPHORESIS 35
3.1.6 PRECIPITATION OF DNA/RNA WITH PHENOL/CHLOROPHORM 35
3.1.7 PRECIPITATION OF DNA 36
3.1.8 PREPARATION OF MRNA FROM TISSUE CULTURE CELLS 36
3.1.9 PREPARATION OF DNA FROM TISSUE OR TISSUE CULTURE CELLS FOR
GENOTYPING 36
3.1.10 POLYMERASE CHAIN REACTION (PCR) 37
3.1.11 CDNA SYNTHESIS / REVERSE TRANSCRIPTION (RT) 37
3.1.12 QUANTITATIVE PCR ANALYSIS 38
3.1.13 DNA SEQUENCING 38
3.1.14 RNA INTERFERENCE 39
3.1.15 CHROMATIN IMMUNOPRECIPITATION ASSAY (CHIP) 39
3.1.16 RIBOPROBE SYNTHESIS FOR IN SITU HYBRIDIZATION 40
3.1.17 IN SITU HYBRIDIZATION 41
3.1.18 CDNA MICROARRAY 42
3.2 PROTEINS 42
3.2.1 PREPARATION OF WHOLE CELL EXTRACTS FROM EUKARYOTIC TISSUE CULTURE
CELLS 42
3.2.2 PREPARATION OF NUCLEAR EXTRACTS FOR MASS SPECTROMETRY 42
3.2.3 DETERMINATION OF PROTEIN CONCENTRATIONS 43
3.2.4 IMMUNOPRECIPITATION 43
3.2.5 MASS SPECTROMETRY OF LSDL-INTERACTING PROTEINS 43
3.2.6 SUCROSE GRADIENT CENTRIFUGATION 44
3.2.7 DENATURING POLYACRYLAMIDE SDS GEL ELECTROPHORESIS (SDS-PAGE) 44
3.2.8 WESTERN BLOTTING AND IMMUNODETECTION 45
3.2.9 IMMUNOFLUORESCENT STAINING OF TISSUE CULTURE CELLS 45
3.2.10 IMMUNOHISTOFLUORESCENCE STAINING OF PARAFFIN EMBEDDED EMBRYO
SECTIONS 46
3.2.11 HE STAINING OF PARAFFIN EMBEDDED EMBRYO SECTIONS 46
3.3 BACTERIA 47
3.3.1 TRANSFORMATION OF E. COLI CELLS 47
3.4 EUKARYOTIC CELL CULTURE
AND MOUSE EXPERIMENTS 47
CONTENTS
3.4.1 PERMANENT CULTURE OF EUKARYOTIC CELL LINES 47
3.4.2 PREPARATION OF MOUSE EMBRYONIC FIBROBLASTS (MEFS) 47
3.4.3 PREPARATION OF MMC-MEFS 48
3.4.4 ISOLATION AND MAINTENANCE OF TROPHOBLAST STEM CELLS (TSCS) 48
3.4.5 CRYO-CONSERVATION OF EUKARYOTIC CELL CULTURE CELLS 49
3.4.6 TRANSIENT TRANSFECTION OF EUKARYOTIC CELL CULTURE CELLS 49
3.4.7 TRANSIENT TRANSFECTION OF TSCS 50
3.4.8 MIGRATION AND INVASION ASSAYS 50
3.4.9 PROLIFERATION ASSAY 51
3.4.10 FACS (FLUORESCENCE-ACTIVATED CELL SORTING) 51
3.4.11 PREPARATION OF PARAFFIN EMBEDDED MOUSE EMBRYOS 51
3.4.12 GENERATION OF TRANSGENIC MICE 52
3.5 COMPUTATIONAL DEVICES 53
3.5.1 DATABASES, COMPUTATIONAL ANALYSES AND ALIGNMENT PROGRAMS 53
3.5.2 SOFTWARES 53
4 RESULTS 54
4.1 DIFFERENTIAL REQUIREMENTS
OF LSDL IN EMBRYONIC TISSUES 54
4.2 DELETION OF LSDL IMPAIRS TROPHECTODERM DEVELOPMENT 55
4.3 LSDL IS DISPENSABLE FOR THE
MAINTENANCE OF THE
STERNNESS STATE 58
4.4 LSDL REGULATES THE DIFFERENTIATION OF TSCS 59
4.5 LSDL IS DISPENSABLE FOR PROLIFERATION OF UNDIFFERENTIATED TSCS 64
4.6 LSDL DETERMINES THE CELL
SIZE OF TSCS 65
4.7 LSDL RESTRAINS MIGRATION AND INVASION
OF TSCS 66
4.8 DELETION OF LSDL LOWERS THE THRESHOLD FOR DIFFERENTIATION
ONSET 68
4.9 LSDL CONTROLS EXPRESSION OF OVOL2 IN TSCS
AND TROPHECTODERM 72
4.10 LSDL DIRECTLY SUPPRESSES OVOL2 EXPRESSION IN TSCS 74
4.11 OVOL2 INDUCES
MIGRATION
AND INVASION
OF LSDL-/-
TSCS 78
4.12 LSDL REGULATES MIGRATION IN TSCS INDEPENDENT
OF THE
MAP3K4
PATHWAY 80
4.13 LSDL INTERACTS WITH A WIDE
RANGE OF PROTEINS 81
4.13.1 ///C2-DEPLETION LEADS TO ENHANCED MIGRATION/INVASION OF
UNDIFFERENTIATED TSCS
85
4.13.2 BRGL AND BRM ARE ESSENTIAL FOR TSC DIFFERENTIATION 86
5 DISCUSSION AND PERSPECTIVES 89
5.1 THE ROLE
OF LSDL IN STEM CELL MAINTENANCE 89
CONTENTS
5.1.1 LSDL CONTROLS DIFFERENTIATION OF STEM CELLS 89
5.1.2 IMPAIRED TROPHOBLAST GIANT CELLS DEVELOPMENT IN
LSDL-/-
TSCS 92
5.2 LSDL REGULATES MIGRATION OF TSCS 93
5.3 SIGNALLING PATHWAYS
IN LSDL-/- TSCS 95
5.4 LSDL IS PART OF MANY
COMPLEXES 98
5.4.1 LSDL-SWI/SNF COMPLEX 99
5.4.2 LSDL INTERACTS WITH HIC2 100
5.5 CONCLUSION 101
TABLES 103
REFERENCES 106
LIST OF ABBREVIATION 121
ACKNOWLEDGEMENTS 124
|
| any_adam_object | 1 |
| author | Hölz, Stefanie |
| author_GND | (DE-588)142117803 |
| author_facet | Hölz, Stefanie |
| author_role | aut |
| author_sort | Hölz, Stefanie |
| author_variant | s h sh |
| building | Verbundindex |
| bvnumber | BV042140480 |
| ctrlnum | (OCoLC)891569931 (DE-599)DNB1055852875 |
| dewey-full | 610 570 |
| dewey-hundreds | 600 - Technology (Applied sciences) 500 - Natural sciences and mathematics |
| dewey-ones | 610 - Medicine and health 570 - Biology |
| dewey-raw | 610 570 |
| dewey-search | 610 570 |
| dewey-sort | 3610 |
| dewey-tens | 610 - Medicine and health 570 - Biology |
| discipline | Biologie Medizin |
| format | Thesis Book |
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| spelling | Hölz, Stefanie Verfasser (DE-588)142117803 aut Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells vorgelegt von Stefanie Hölz 2014 124 Bl. Ill., graph. Darst. 30 cm txt rdacontent n rdamedia nc rdacarrier Freiburg (Breisgau), Univ., Diss., 2014 (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=027580457&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Hölz, Stefanie Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells |
| subject_GND | (DE-588)4113937-9 |
| title | Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells |
| title_auth | Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells |
| title_exact_search | Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells |
| title_full | Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells vorgelegt von Stefanie Hölz |
| title_fullStr | Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells vorgelegt von Stefanie Hölz |
| title_full_unstemmed | Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells vorgelegt von Stefanie Hölz |
| title_short | Lysine-specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells |
| title_sort | lysine specific demethylase 1 regulates differentiation onset and migration of trophoblast stem cells |
| topic_facet | Hochschulschrift |
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