Identification of novel molecular factors involved in individual stress vulnerability:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
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| Schlagworte: | |
| Online-Zugang: | kostenfrei http://d-nb.info/1029040397/34 https://nbn-resolving.org/urn:nbn:de:bvb:19-150881 Inhaltsverzeichnis |
| Beschreibung: | IV, 151 S. Ill., graph. Darst. |
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| 245 | 1 | 0 | |a Identification of novel molecular factors involved in individual stress vulnerability |c Sebastian Scharf |
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Datensatz im Suchindex
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|---|---|
| adam_text | IMAGE 1
CONTENTS
C O N T E N T S
1 I N T R O D U C T I O N 1
1.1 THE CONCEPT OF STRESS 1
1.2 THE STRESS RESPONSE 1
1.2.1 THE SYMPATHETIC NERVOUS SYSTEM 1
1.2.2 THE HYPOTHALAMIC-PITUITARY-ADRENAL AXIS 2
1.3 MODULATION OF THE HP A AXIS 5
1.3.1 THE ROAD TO REGULATION: CONNECTIVITY OF THE PVN . . . 5
1.3.2 ACTIVATING INFLUENCES 5
1.3.3 INHIBITORY INFLUENCES AND FEEDBACK 6
1.3.4 CIRCADIAN RHYTHM AND PULSATILITY 7
1.4 ACUTE VERSUS CHRONIC STRESS 7
1.5 STRESS AS A RISK FACTOR FOR DISEASE 8
1.6 IMPORTANT TIME WINDOWS FOR STRESS EXPOSURE AND DEVELOPMENT 9
1.6.1 THE PRENATAL PERIOD 9
1.6.2 THE POSTNATAL PERIOD 10
1.6.3 ADOLESCENCE 11
1.6.4 ADULTHOOD 11
1.6.5 AGING 12
1.7 THE PERSISTENCE OF STRESS EFFECTS 12
1.8 THE CONCEPT OF VULNERABILITY AND RESILIENCE 12
1.9 MODELS OF VULNERABILITY AND RESILIENCE 13
1.9.1 TRANSGENIC MODELS 13
1.9.2 SELECTION OF SUBPOPULATIONS 14
1.9.3 SELECTIVE BREEDING 16
1.10 THE EVOLUTIONARY PERSPECTIVE OF VULNERABILITY 16
1.11 THE HIPPOCAMPUS AS A PRIME TARGET IN STRESS RESEARCH 17
1.12 MOUSE MODELS IN PSYCHIATRIC RESEARCH 17
1.13 ACTIVITY-REGULATED CYTOSKELETON-ASSOCIATED PROTEIN 18
1.13.1 INDUCTION OF THE ARC GENE 20
1.13.2 STRUCTURE AND BINDING MOTIFS OF THE ARC GENE 20
1.13.3 ARC MRNA TRANSPORT AND POST-TRANSCRIPTIONAL MODIFICATIONS 21
1.13.4 TRANSLATIONAL CONTROL OF ARC 21
1.13.5 FUNCTIONAL RELEVANCE OF ARC 22
1.13.5.1 ELECTROPHYSIOLOGY 22
1.13.5.2 LEARNING AND MEMORY 22
1.13.5.3 STRESS AND STRESS VULNERABILITY 23
1.14 AIMS 23
2 M A T E R I A L S & M E T H O D S 2 4
2.1 ANIMALS 24
2.2 EXPERIMENTAL DESIGN 24
2.2.1 EXPERIMENT 1: LONG-TERM EFFECTS OF STRESS EXPOSURE ON GENE
TRANSCRIPTION IN THE HIPPOCAMPUS 24
2.2.1.1 EXPERIMENT LA: LONG-TERM EFFECTS FOLLOWING CHRONIC SOCIAL STRESS
EXPOSURE 24
2.2.1.2 EXPERIMENT LB: THE EFFECT OF ACUTE STRESS EXPOSURE ON SELECTED
CANDIDATE GENES 25
2.2.1.3 EXPERIMENT LC: INFLUENCE OF THE GR ON SELECTED GENES VIA PHARMA
COLOGICAL ACTIVATION 25
I
HTTP://D-NB.INFO/1030445516
IMAGE 2
CONTENTS
2.2.2 EXPERIMENT 2: TRANSCRIPTOME DIFFERENCES BETWEEN STRESS-VULNERABLE
AND STRESS-RESILIENT ANIMALS 29
2.2.2.1 EXPERIMENT 2A: VULNERABILITY GENES IN THE HIPPOCAMPUS 29 2.2.2.2
EXPERIMENT 2B: VULNERABILITY GENES IN PERIPHERAL LYMPHOCYTES . . 29
2.2.3 EXPERIMENT 3: CAUSAL INFLUENCE OF ARC ON STRESS VULNERABILITY 30
2.2.3.1 EXPERIMENT 3A: EFFICACY OF THE ARC SHRNA KNOCKDOWN 30
2.2.3.2 EXPERIMENT 3B: INFLUENCE OF ARC SHRNA KNOCKDOWN IN THE
HIPPOCAMPAL CA1 REGION ON STRESS VULNERABILITY 30
2.3 STRESS PARADIGMS 31
2.3.1 CHRONIC SOCIAL STRESS 31
2.3.2 CHRONIC SOCIAL DEFEAT 31
2.3.3 RESTRAINT STRESS 32
2.3.4 FOOD DEPRIVATION 32
2.4 SAMPLING PROCEDURE 32
2.5 CARDIAC PERFUSION 32
2.6 RADIOIMMUNOASSAY 33
2.7 BEHAVIOURAL TESTS 33
2.7.1 OPEN FIELD TEST 33
2.7.2 OBJECT RECOGNITION TEST 34
2.7.3 SOCIABILITY TEST 34
2.7.4 FORCED SWIM TEST & STRESS RESPONSE TEST 35
2.8 BRAIN PROCESSING 35
2.9 LASER-MICRODISSECTION 36
2.10 RNA ISOLATION 37
2.10.1 WHOLE HIPPOCAMPI 37
2.10.2 LASER-MICRODISSECTION 37
2.11 RNA QUALITY CONTROL 37
2.12 RNA AMPLIFICATION 38
2.13 CDNA TRANSCRIPTION 38
2.14 MICROARRAY ANALYSIS 39
2.15 QUANTITAVE RT-PCR 39
2.16 IN-SITU HYBRIDISATION 41
2.17 PROTEIN ISOLATION 42
2.18 WESTERN BLOT 42
2.19 IMMUNOFLUORESCENCE 43
2.20 KNOCKDOWN VIRUS OF ARC 43
2.21 STEREOTACTIC SURGERY 43
2.22 STATISTICAL ANALYSES 44
3 R E S U L T S 4 5
3.1 EXPERIMENT 1: LONG-TERM EFFECTS OF STRESS EXPOSURE ON GENE
TRANSCRIPTION IN THE HIPPOCAMPUS 45
3.1.1 EXPERIMENT LA: LONG-TERM EFFECTS FOLLOWING CHRONIC SOCIAL STRESS
EXPOSURE . 45 3.1.1.1 VALIDITY OF THE STRESS PARADIGM 45
3.1.1.2 RNA QUALITY 45
3.1.1.3 MICROARRAY ANALYSIS 47
3.1.2 VALIDATION VIA QRT-PCR IN THE REPLICATION SAMPLE 49
3.1.2.1 VALIDITY OF THE STRESS PARADIGM 49
3.1.2.2 RNA QUALITY 49
3.1.2.3 QUALITY CONTROL OF THE QRT-PCR 51
3.1.2.4 RESULTS FROM THE QRT-PCR 52
II
IMAGE 3
CONTENTS
3.1.3 VALIDATION VIA IN-SITU HYBRIDISATION 56
3.1.3.1 MTAPL 56
3.1.3.2 FANCM 56
3.1.3.3 AKAP8 56
3.1.3.4 IFFOL 56
3.1.3.5 RUFY3 56
3.1.4 EXPERIMENT LB: THE EFFECT OF ACUTE STRESS EXPOSURE ON SELECTED
CANDIDATE GENES 58
3.1.4.1 IFFOL 58
3.1.5 EXPERIMENT LC: INFLUENCE OF THE GR ON SELECTED GENES VIA
PHARMACOLOGICAL ACTIVATION 59
3.1.5.1 IFFOL 59
3.2 EXPERIMENT 2: TRANSCRIPTOME DIFFERENCES BETWEEN STRESS-VULNERABLE
AND STRESSRESILIENT ANIMALS 61
3.2.1 VULNERABILITY GENES IN THE HIPPOCAMPUS 61
3.2.1.1 EXPERIMENT 2A: MICROARRAY OF THE DISCOVERY SAMPLE 61
3.2.1.1.1 VALIDITY OF THE STRESS PARADIGM 61
3.2.1.1.2 RNA QUALITY 61
3.2.1.1.3 MICROARRAY ANALYSIS 61
3.2.1.2 VALIDATION VIA QRT-PCR IN THE REPLICATION SAMPLE 63
3.2.1.2.1 VALIDITY OF THE STRESS PARADIGM 63
3.2.1.2.2 RNA QUALITY 63
3.2.1.2.3 RESULTS FROM THE QRT-PCR 63
3.2.2 EXPERIMENT 2B: VULNERABILITY GENES IN PERIPHERAL LYMPHOCYTES 63
3.2.2.1 MICROARRAY OF THE DISCOVERY SAMPLE 63
3.2.2.1.1 VALIDITY OF THE STRESS PARADIGM 63
3.2.2.1.2 RNA QUALITY 65
3.2.2.1.3 RESULTS FROM THE MICROARRAY 65
3.2.2.2 VALIDATION VIA QRT-PCR IN THE DISCOVERY SAMPLE 65
3.2.2.2.1 VALIDITY OF THE STRESS PARADIGM 65
3.2.2.2.2 RNA QUALITY 65
3.2.2.2.3 RESULTS FROM THE QRT-PCR 65
3.2.3 CORRELATIONS BETWEEN BLOOD AND BRAIN 66
3.2.3.1 COMBINED CORRELATIONS 66
3.2.3.2 CORRELATIONS WITHIN THE SUBGROUPS 66
3.3 EXPERIMENT 3: CAUSAL INFLUENCE OF ARC ON STRESS VULNERABILITY 71
3.3.1 EXPERIMENT 3A: CONFIRMATION OF THE ARC KNOCKDOWN 71
3.3.1.1 IN-SITU HYBRIDISATION 71
3.3.1.2 WESTERN BLOT 71
3.3.1.3 IMMUNFLUORESCENCE 71
3.3.2 EXPERIMENT 3B: INFLUENCE OF ARC KNOCKDOWN IN THE HIPPOCAMPUS ON
STRESS VULNERABILITY 74
3.3.2.1 LOCALISATION 74
3.3.2.2 BODY WEIGHT 76
3.3.2.3 FUR STATE 77
3.3.2.4 CORTICOSTERONE 78
3.3.2.5 ADRENAL GLANDS 79
3.3.2.6 BEHAVIOUR IN THE LAST WEEK OF SOCIAL DEFEAT 80
3.3.2.6.1 OPEN FIELD TEST 80
3.3.2.6.2 OBJECT RECOGNITION TEST 83
3.3.2.6.3 SOCIABILITY TEST 84
III
IMAGE 4
CONTENTS
3.3.2.6.4 FORCED SWIM TEST 85
3.3.2.7 BEHAVIOUR DIRECTLY AFTER THE SOCIAL DEFEAT 87
3.3.2.7.1 SUCROSE PREFERENCE TEST 87
3.3.2.8 BEHAVIOUR AFTER RECOVERY 88
3.3.2.8.1 OPEN FIELD TEST 88
3.3.2.8.2 OBJECT RECOGNITION TEST 90
3.3.2.8.3 SOCIABILITY TEST 90
3.3.2.8.4 FORCED SWIM TEST 91
4 D I S C U S S I O N 9 3
4.1 PART 1: LONG-TERM EFFECTS OF STRESS EXPOSURE ON GENE TRANSCRIPTION
IN THE HIPPOCAMPUS 93 4.1.1 MICROARRAY - THE PITFALLS OF WHOLE GENOME
EXPRESSION STUDIES 93
4.1.2 GENE EXPRESSION CHANGES FOLLOWING CHRONIC STRESS EXPOSURE 94
4.1.3 SUBGROUPS WITHIN THE STRESSED PHENOTYPES 94
4.1.4 IFFOL - A CANDIDATE FOR FUTURE STUDIES? 95
4.1.5 CONCLUSIONS 96
4.2 PART 2: TRANSCRIPTOME DIFFERENCES BETWEEN STRESS-VULNERABLE AND
STRESS-RESILIENT ANIMALS 97
4.2.1 DIFFERENCES IN THE HIPPOCAMPUS 97
4.2.2 DIFFERENCES IN THE BLOOD 98
4.2.3 CORRELATIONS BETWEEN BRAIN AND BLOOD 99
4.2.4 VULNERABILITY OR CORT RESPONSE? 101
4.2.5 ARC AS A CANDIDATE 101
4.2.6 CONCLUSIONS 101
4.3 PART 3: CAUSAL INFLUENCE OF ARC ON STRESS VULNERABILITY 102
4.3.1 INVESTIGATION OF CAUSALITY - A NEW DESIGN FOR A NEW RESEARCH
QUESTION . . . . 102 4.3.2 ARC KNOCKDOWN PARTIALLY PREVENTS
STRESS-INDUCED CHANGES IN BEHAVIOUR . . . 103 4.3.3 ARC KNOCKDOWN DOES
NOT INFLUENCE BASIC PHYSIOLOGICAL PARAMETERS 104 4.3.4 POSSIBLE
MOLECULAR MECHANISMS OF ARC IN STRESS VULNERABILITY 105 4.3.5 THE
POSSIBLE EVOLUTIONARY JUSTIFICATION OF ARC 107
4.3.6 CONCLUSIONS 108
4.4 SYNOPSIS 108
4.5 FUTURE DIRECTIONS 109
5 S U M M A R Y 1 1 0
BIBLIOGRAPHY 1 1 1
LIST O F F I G U R E S 1 3 5
LIST O F T A B L E S 1 3 7
LIST O F A B B R E V A T I O N S 1 3 9
CURRICULUM V I T A E 1 4 5
P U B L I C A T I O N RECORD 1 4 7
A C K N O W L E D G E M E N T S 1 4 9
A S S E R T A T I O N (ERKLARUNG) 1 5 1
IV
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| spelling | Scharf, Sebastian Verfasser aut Identification of novel molecular factors involved in individual stress vulnerability Sebastian Scharf 2012 IV, 151 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2012 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-150881 http://edoc.ub.uni-muenchen.de/15088/ Verlag kostenfrei Volltext http://d-nb.info/1029040397/34 Langzeitarchivierung Nationalbibliothek https://nbn-resolving.org/urn:nbn:de:bvb:19-150881 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025871112&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Scharf, Sebastian Identification of novel molecular factors involved in individual stress vulnerability |
| subject_GND | (DE-588)4113937-9 |
| title | Identification of novel molecular factors involved in individual stress vulnerability |
| title_auth | Identification of novel molecular factors involved in individual stress vulnerability |
| title_exact_search | Identification of novel molecular factors involved in individual stress vulnerability |
| title_full | Identification of novel molecular factors involved in individual stress vulnerability Sebastian Scharf |
| title_fullStr | Identification of novel molecular factors involved in individual stress vulnerability Sebastian Scharf |
| title_full_unstemmed | Identification of novel molecular factors involved in individual stress vulnerability Sebastian Scharf |
| title_short | Identification of novel molecular factors involved in individual stress vulnerability |
| title_sort | identification of novel molecular factors involved in individual stress vulnerability |
| topic_facet | Hochschulschrift |
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