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Language(s):
eng - English
Dates:
2009-03-27
Publication Status:
Accepted / In Press
Pages:
[1], XIV, 79 S. S. A - Z, AA - CC
Publishing info:
Berlin : Freie Universität Berlin
Table of Contents:
Contents
ABBREVIATIONS V
LIST OF FIGURES VII
LIST OF TABLES IX
ABSTRACT XI
ABSTRACT (GERMAN) XIII
1. INTRODUCTION 1
1.1 Aging 1
1.1.1 Aging theories 1
1.1.2 Model systems in aging research 5
1.1.3 Human skin aging 9
1.2 Whole genome microarray analyses 10
1.2.1 Molecular methods employed for gene expression analyses 10
1.2.2 Advantages and disadvantages of whole genome studies 11
1.2.3 Gene expression microarrays and aging 12
1.3 Aim of this work 13
2. MATERIAL AND METHODS 14
2.1 Model systems 14
2.1.1 Cell cultures 14
2.1.2 Human skin biopsies 14
2.1.3 Mice 15
2.2 RNA analyses 15
2.2.1 Total RNA isolation 15
2.2.2 RNA and cDNA quantification 16
2.2.3 Agarose gel electrophoresis 17
2.2.4 Reverse transcription 17
2.2.5 Real-time polymerase chain reaction (Real-Time PCR) 18
2.2.6 ENSEMBL chip hybridisation 18
2.2.7 Illumina bead chip hybridisation 20
2.2.8 Pathway analysis 21
2.3 Protein analyses 21
2.3.1 Protein isolation 21
2.3.2 Protein quantification (Bradford) 22
2.3.3 SDS-PAGE gel electrophoresis 22
2.3.4 Western blotting 23
2.4 Metabolite analyses 24
2.4.1 Lipid hydroperoxide (LPO) measurement 24
2.5 Histology 24
2.5.1 Section preparation 24
2.5.2 Hematoxylin Eosin staining 25
3. RESULTS 27
3.1 Global data analysis 27
3.1.1 Global gene expression analysis 27
3.1.2 RNA quality control 29
3.1.3 Target gene lists 30
3.1.4 Pathway analyses 31
3.1.5 Array data confirmation by Real-Time PCR 31
3.2 Human skin aging 33
3.2.1 Genetic changes in age-dependent hormone-treated cell cultures 33
3.2.2 Gene expression in young and old sun-protected skin biopsies 35
3.2.3 Analogous gene expression of in vitro and in vivo skin samples 38
3.3 Mouse aging (brain, heart & kidney) 40
3.3.1 Synchronizing the sample collection 40
3.3.2 Physical and morphological differences of young and aged mice 45
3.3.3 Age-dependent gene expression in mouse brain, heart and kidney 46
3.3.4 Age-related post-transcriptional and structural changes 53
3.4 Comparison of human and mouse aging 55
3.4.1 Correlation of age-regulated target genes 55
3.4.2 Correlation of age-regulated Kegg pathways and Gene Ontologies (BP and
CC) 57
3.4.3 Correlation of target genes to published data 62
4. DISCUSSION 64
4.1 Global data analysis 64
4.2 Human skin aging 67
4.3 Mouse aging 72
4.4 The basis of aging 76
5. CONCLUSION 78
REFERENCES A
PUBLICATIONS L
CURRICULUM VITAE M
APPENDIX N
I Buffers for SDS-PAGE gel electrophoresis N
II Buffers for western blotting O
III Target gene lists P
IV David output tables W
Rev. Type:
-
Identifiers:
eDoc: 456429
Degree:
PhD