Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
|
| Schlagworte: | |
| Online-Zugang: | Volltext Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in dt. und engl. Sprache |
| Beschreibung: | 164 S. graph. Darst. |
Internformat
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| 100 | 1 | |a Araragi, Naozumi |d 1982- |e Verfasser |0 (DE-588)1043403930 |4 aut | |
| 245 | 1 | 0 | |a Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice |c von Naozumi Araragi |
| 246 | 1 | 3 | |a Elektrophysiologische Untersuchung bei zwei Tiermodellen für emotionale Störungen - Serotonin Transporter knockout Mäuse und Tryptophan Hydroxylase 2 knockout Mäuse |
| 264 | 1 | |c 2013 | |
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| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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Datensatz im Suchindex
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| adam_text | TABLE OF CONTENTS
TABLE OF CONTENTS
AFFIDAVIT 3
EIDESSTATTLICHE ERKLARUNG 3
ABSTRACT 4
ZUSAMMENFASSUNG 5
1 . INTRODUCTION 14
1.1. SEROTONIN AND AFFECTIVE DISORDERS 14
1.1.1. DEPRESSION AND MONOAMINE NEUROTRANSMITTERS 14
1.1.2. DEPRESSION AND ANXIETY DISORDERS 15
1.1.3. ANIMAL MODELS OF DEPRESSION AND ANXIETY DISORDERS 16
1.2. SEROTONIN AND SEROTONERGIC NEURONS 17
1.2.1. SYNTHESIS AND METABOLISM 17
1.2.2. ANATOMY OF BRAIN SEROTONIN SYSTEM 18
1.2.3. SEROTONERGIC NEURONS 20
I). ELECTROPHYSIOLOGICAL SIGNATURE 20
II). SPONTANEOUS FIRING 20
III). AUTOINHIBITION OF SEROTONERGIC NEURON FIRING 22
IV). SEROTONIN RELEASE 23
V). MECHANISM OF 5-HT
IA
RECEPTOR ACTION 24
1.2.4. THE DRN AND EMOTION REGULATION 25
1.3. SEROTONIN TRANSPORTER (5-HTT) AND ITS ASSOCIATION WITH EMOTIONAL
DISORDERS 26
1.4. SEROTONIN TRANSPORTER KNOCKOUT (5-HTT KO) MICE AS A MODEL FOR
DEPRESSION-RELATED
EMOTIONALITY 28
1.4.1. CHARACTERISTICS OF 5-HTT KO MICE 28
1.4.2. ELECTROPHYSIOLOGY 31
I). IN VITRO ELECTROPHYSIOLOGY 31
II). IN VIVO ELECTROPHYSIOLOGY 32
1.5. HIPPOCAMPUS AND DEPRESSION 33
1.5.1. MEMORY AND DEPRESSION 33
1.5.2. HIPPOCAMPUS AND STRESS 35
7
HTTP://D-NB.INFO/1043539050
TABLE OF CONTENTS
1.5.3. PHARMACOLOGIC INTERVENTIONS ON THE HIPPOCAMPUS -ANTIDEPRESSANTS,
ANXIOLYTICS,
AND DRUGS OF ABUSE~ 36
1.5.4. 5-HT RECEPTORS IN THE HIPPOCAMPUS 37
I). 5-HT
IA
RECEPTORS 38
II). 5-HT3 RECEPTORS 39
III). 5
-HT7
RECEPTORS 40
1.5.5. EFFECTS OF SEROTONIN ON LEARNING AND SYNAPTIC PLASTICITY 41
1.6. TRYPTOPHAN-HYDROXYLASE 2 (TPH2) AND TPH2 KO MICE 44
1.6.1. DISCOVERY OF TWO ISOFORMS OF TPH AND GENERATION OF TPH2 KO MICE
44
1.6.2. CHARACTERISTICS OF TPH2 KO MICE 45
1.6.3. ASSOCIATION OF TPH2 WITH NEUROPSYCHIATRIC DISORDERS 47
1.7. AIMS OF THE STUDY 48
2 . MATERIALS AND METHODS 50
2.1. ANIMAL HANDLING AND STRAINS 50
2.2. GENOTYPING OF MICE 50
2.2.1. TAIL DIGESTION AND DNA PURIFICATION 50
2.2.2. PCR 51
I). 5-HTT 51
II). TPH2 52
2.2.3. ELECTROPHORESIS 53
2.3. LOOSE-SEAL CELL-ATTACHED RECORDING 54
2.3.1. RATIONALE 54
2.3.2. PREPARATION OF DORSAL RAPHE SLICES 54
2.3.3. ELECTROPHYSIOLOGICAL RECORDING 55
2.3.4. DRUGS 56
2.3.5. IDENTIFICATION OF SEROTONERGIC NEURONS 56
I). MORPHOLOGICAL & HISTOLOGICAL CRITERIA 56
II). ELECTROPHYSIOLOGICAL CRITERIA 56
2.3.6. QUALITY CONTROL 57
I). AMOUNT OF INTER-SPIKE CURRENT (ISC) 57
8
TABLE OF CONTENTS
II). ACTION CURRENT SHAPE SHOULD REMAIN THE SAME 58
III). ACTION CURRENT AMPLITUDE SHOULD BE MONITORED TO AVOID FALSE
POSITIVE EFFECTS OF
DRUGS 59
IV). IHOID
SHOULD BE MONITORED TO PREVENT PIPETTE CURRENT FROM AFFECTING FIRING
FREQUENCY 60
2.4. WHOLE-CELL (WC) RECORDING 61
2.4.1. RATIONALE 61
I). DORSAL RAPHE NUCLEUS RECORDING 62
2.4.2. MATERIALS AND METHODS 62
I). ANIMALS 62
II). PREPARATION OF DORSAL RAPHE NUCLEUS SLICES 62
III). ELECTROPHYSIOLOGICAL RECORDING 62
2.4.3. PARAMETERS MEASURED 63
I). RESTING MEMBRANE POTENTIAL (V
M
) 63
II). SERIES RESISTANCE (R
S
), MEMBRANE RESISTANCE (R
M
), CELL CAPACITANCE (C
M
), TIME
CONSTANT (
T
) 63
III). ACTION POTENTIAL (AP) SHAPE (AMPLITUDE, HHW, X OF AHP) 65
IV). MAXIMUM FIRING RATE 67
V). SAG AMPLITUDE 68
2.4.4. IDENTIFICATION OF SEROTONERGIC NEURONS IN THE WC RECORDING
CONFIGURATION 69
I). MORPHOLOGICAL & HISTOLOGICAL CRITERIA 69
II). ELECTROPHYSIOLOGICAL CRITERIA 69
2.4.5. QUALITY CONTROL 70
I). THE SERIES RESISTANCE SHOULD NOT EXCEED ~20 MQ THROUGHOUT THE
RECORDING 70
II). THE MEMBRANE POTENTIAL SHOULD B.E MORE NEGATIVE THAN -50 MV 71
III). MEMBRANE CAPACITANCE AND RESISTANCE MUST BE STABLE AT THE STEADY
STATE 71
II). HIPPOCAMPUS RECORDING 71
2.4.6. ANIMALS 71
2.4.7. PREPARATION OF HIPPOCAMPAL SLICES 71
2.4.8. ELECTROPHYSIOLOGICAL RECORDING 72
2.4.9. STAINING PROCEDURES 73
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TABLE OF CONTENTS
2.5. EXTRACELLULAR FIELD POTENTIAL RECORDING 73
2.5.1. RATIONALE 73
2.5.2. PREPARATION OF HIPPOCAMPAL SLICES 74
2.5.3. ELECTROPHYSIOLOGICAL RECORDING 74
2.5.4. QUALITY CONTROL OF LTP DATA 76
2.6. A DOSE-RESPONSE CURVE AND EC50 77
2.6.1. RATIONALE 77
I). A DOSE-RESPONSE CURVE 77.
II).
EC50
78
2.6.2. PROCEDURE 78
2.7. STATISTICS 79
3 . RESULTS 81
3.1. LOOSE-SEAL CELL-ATTACHED RECORDING FROM SEROTONERGIC NEURONS IN THE
DRN OF 5-HTT AND
TPH2 KO MICE (STUDY 1) 81
3.1.1. RATIONALE 81
3.1.2. IDENTIFICATION OF SEROTONERGIC NEURONS 81
3.1.3. COMPARISON OF BASELINE FIRING RATES 83
3.1.4. COMPARISON OF UDI AMONG GENOTYPES 84
3.1.5. ASSESSMENT OF 5-HT
IA
RECEPTOR FUNCTION BY A SELECTIVE AGONIST R8HD 85
3.1.6. TRYPTOPHAN: TESTING 5-HT SYNTHESIS BY APPLYING A 5-HT PRECURSOR
88
I). COMPLETE ABSENCE OF 5-HT SYNTHESIS IN TPH2 KO MICE (PUBLISHED IN
GUTKNECHT ET
AL., 2012) 88
II). COMPARISON OF TRP-DRIVEN 5-HT SYNTHESIS AND AUTOINHIBITION AMONG
GENOTYPES .. 89
3.1.7. TEST OF 5-HT SYNTHESIS FROM 5-HTP 91
3.2. WC RECORDING FROM SEROTONERGIC NEURONS IN THE DRN OF TPH2 KO MICE
(STUDY 2) 93
3.2.1. STATIC MEMBRANE PROPERTIES 93
I). RESTING MEMBRANE POTENTIAL 93
II). MEMBRANE RESISTANCE AND CAPACITANCE 94
III). MEMBRANE CONDUCTANCE 95
IV). SERIES RESISTANCE AND HOLDING CURRENT 96
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V). SAG AMPLITUDE . 97
3.2.2. ACTIVE MEMBRANE PROPERTIES 98
I). ACTION POTENTIAL PROPERTIES 98
II). CONDUCTANCE IN RESPONSE TO VOLTAGE RAMP 99
3.3. WC RECORDING FROM HIPPOCAMPAL CA1 PYRAMIDAL NEURONS IN 5-HTT KO
MICE (STUDY 3)
100
3.3.1. STATIC MEMBRANE PROPERTIES 100
3.3.2. ACTION POTENTIAL PROPERTIES 101
3.3.3. SYNAPTIC TRANSMISSION 103
3.3.4.1/V RELATIONSHIP 104
3.3.5. MORPHOLOGICAL ANALYSIS 106
3.4. EXTRACELLULAR FIELD POTENTIAL RECORDING OF LONG-TERM POTENTIATION
(LTP) (STUDY 4) 108
3.4.1. TIME COURSE OF FEPSP CHANGE 108
3.4.2. SHAPE OF FEPSP 108
3.4.3. COMPARISON OF NORMALISED FEPSP ACROSS DIFFERENT GENOTYPES 110
3.4.4. SUPERPOSITION OF FEPSP TRACES AND COMPARISON OF % POTENTIATION
111
4 . DISCUSSION 113
4.1. LOOSE-SEAL CELL-ATTACHED RECORDING FROM SEROTONERGIC NEURONS IN THE
DRN OF 5-HTT AND
TPH2 KO MICE (STUDY 1) 113
4.1.1. BASELINE PACEMAKER FIRING ACTIVITY WAS PRESERVED IN KO MICE 113
4.1.2. SEROTONERGIC NEURONS IN TPH2 KO MICE AND 5-HTT KO MICE RETAIN
NORMAL
ELECTROPHYSIOLOGICAL PROPERTIES 113
4
.1
.3. RESPONSIVENESS OF 5-HT
JA
RECEPTORS WERE SHIFTED IN THE OPPOSITE DIRECTION IN TPH2
KO AND 5-HTT KO MICE 114
4.1.4. LACK OF 5-HT SYNTHESIS FROM TRP IN TPH2 KO MICE AND PRESENCE OF
AUTOINHIBITION
IN 5-HTT KO MICE 116
4.1.5. SEROTONERGIC NEURONS OF TPH2 KO AND 5-HTT KO MICE ARE MORE
RESPONSIVE TO 5-
HTP THAN THOSE OF WILDTYPES 117
4.1.6. 5-HT SYNTHESIS AND AUTOINHIBITION 119
4.1.7. DESENSITISATION OR DOWNREGULATION? 120
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TABLE OF CONTENTS
4.1.8. INTERACTION BETWEEN 5-HT
IA
RECEPTORS, 5-HTT, AND OTHER 5-HT RECEPTORS 121
4.1.9. DESENSITISATION OF 5-HT
IA
AUTORECEPTOR IS NOT NECESSARILY ASSOCIATED WITH
BEHAVIOURAL CHANGES BUT MAY BE SECONDARY TO THE INCREASED EXTRACELLULAR
5-HT 123
4.1.10. 5-HT
IA
RECEPTOR SENSITIVITY AND ITS FUNCTIONAL CONSEQUENCES 124
4.1.11. CONCLUSION FOR STUDY 1 125
4.2. WC RECORDING FROM SEROTONERGIC NEURONS IN THE DRN OF TPH2 KO MICE
(STUDY 2)... 127
4.2.1. MEMBRANE RESISTANCE WAS DECREASED IN NEURONS OF KO MICE 127
4.2.2. NEURONS FROM KO MICE HAD LARGER INWARD-RECTIFYING CURRENT IN
RESPONSE TO
NEGATIVE VOLTAGE RAMP 128
4.3. WC RECORDING FROM HIPPOCAMPAL CA1 PYRAMIDAL NEURONS IN 5-HTT KO
MICE (STUDY 3)
129
4.3.1. CA1 PYRAMIDAL NEURONS FROM 5-HTT KO MICE HAD INCREASED
CONDUCTANCE 129
4.3.2. NON-5-HTT MEDIATED SEROTONIN TRANSPORT MAY HAVE COMPENSATED FOR
THE LACK OF 5-
HTT AND CONTRIBUTED TO THE INCREASED CONDUCTANCE IN CA1 NEURONS OF 5-HTT
KO MICE 130
I). OCT3 130
II). PMAT 131
4.3.3. 5-HT3 RECEPTORS MAY ALSO HAVE CONTRIBUTED TO THE INCREASED
CONDUCTANCE 132
4.3.4. MORPHOLOGICAL ANALYSIS 134
4.4. EXTRACELLULAR FIELD POTENTIAL RECORDING OF LTP (STUDY 4) 135
4.4.1. LTP WAS POSSIBLY MAINTAINED BY INTERACTION WITH OTHER
NEUROTRANSMITTERS 135
4.4.2. CHANGES IN LTP INDUCIBILITY IN 5-HTT KO AND TPH2 KO MICE MAY
BECOME EVIDENT
IN PARTICULAR BEHAVIOURAL PARADIGMS 136
4.4.3. ALTERATIONS IN LEARNING ABILITIES OF 5-HTT KO MICE AND TPH2 KO
MICE ARE NOT
CORRELATED WITH CHANGES IN SCHAFFER-COLLATERAL- CA1 LTP 137
4.5. IMPACTS OF CHANGES IN THE 5-HT LEVEL COULD HAVE BEEN BUFFERED BY
AS-YET-UNKNOWN
BIOLOGICAL COMPENSATORY MECHANISMS 138
4.5.1. TPH2KO MICE 138
I). RESIDUAL SEROTONIN 138
II). UPREGULATION OF 5-HT
IA
RECEPTORS 139
III). INTERACTION WITH OTHER NEUROTRANSMITTERS 140
4.5.2. 5-HTT KO MICE 141
12
TABLE OF CONTENTS
CONCLUSION 142
ACKNOWLEDGEMENTS 143
REFERENCES : 144
APPENDICES 157
APPENDIX A. LIST OF FIGURES 157
APPENDIX B. LIST OF FREQUENTLY USED ABBREVIATIONS 161
APPENDIX C. LIST OF PUBLICATIONS 162
APPENDIX D. CURRICULUM VITAE 163
13
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| publishDateSort | 2013 |
| record_format | marc |
| spelling | Araragi, Naozumi 1982- Verfasser (DE-588)1043403930 aut Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice von Naozumi Araragi Elektrophysiologische Untersuchung bei zwei Tiermodellen für emotionale Störungen - Serotonin Transporter knockout Mäuse und Tryptophan Hydroxylase 2 knockout Mäuse 2013 164 S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. und engl. Sprache Würzburg, Univ., Diss., 2013 Maus (DE-588)4169148-9 gnd rswk-swf Elektrophysiologie (DE-588)4138132-4 gnd rswk-swf Knockout Molekulargenetik (DE-588)4671699-3 gnd rswk-swf Serotonin (DE-588)4054614-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Elektrophysiologie (DE-588)4138132-4 s Serotonin (DE-588)4054614-7 s Knockout Molekulargenetik (DE-588)4671699-3 s Maus (DE-588)4169148-9 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-83265 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-83265 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026829512&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Araragi, Naozumi 1982- Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice Maus (DE-588)4169148-9 gnd Elektrophysiologie (DE-588)4138132-4 gnd Knockout Molekulargenetik (DE-588)4671699-3 gnd Serotonin (DE-588)4054614-7 gnd |
| subject_GND | (DE-588)4169148-9 (DE-588)4138132-4 (DE-588)4671699-3 (DE-588)4054614-7 (DE-588)4113937-9 |
| title | Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice |
| title_alt | Elektrophysiologische Untersuchung bei zwei Tiermodellen für emotionale Störungen - Serotonin Transporter knockout Mäuse und Tryptophan Hydroxylase 2 knockout Mäuse |
| title_auth | Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice |
| title_exact_search | Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice |
| title_full | Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice von Naozumi Araragi |
| title_fullStr | Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice von Naozumi Araragi |
| title_full_unstemmed | Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice von Naozumi Araragi |
| title_short | Electrophysiological investigation of two animal models for emotional disorders - serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice |
| title_sort | electrophysiological investigation of two animal models for emotional disorders serotonin transporter knockout mice and tryptophan hydroxylase 2 knockout mice |
| topic | Maus (DE-588)4169148-9 gnd Elektrophysiologie (DE-588)4138132-4 gnd Knockout Molekulargenetik (DE-588)4671699-3 gnd Serotonin (DE-588)4054614-7 gnd |
| topic_facet | Maus Elektrophysiologie Knockout Molekulargenetik Serotonin Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-83265 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026829512&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT araraginaozumi electrophysiologicalinvestigationoftwoanimalmodelsforemotionaldisordersserotonintransporterknockoutmiceandtryptophanhydroxylase2knockoutmice AT araraginaozumi elektrophysiologischeuntersuchungbeizweitiermodellenfuremotionalestorungenserotonintransporterknockoutmauseundtryptophanhydroxylase2knockoutmause |