Structural and functional diversity of synapses in the drosophila CNS: = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
|
| Schlagworte: | |
| Online-Zugang: | Volltext Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in dt. und engl. Sprache |
| Beschreibung: | 186 S. Ill., graph. Darst. |
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| 245 | 1 | 0 | |a Structural and functional diversity of synapses in the drosophila CNS |b = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila |c von Till Felix Malte Andlauer |
| 246 | 1 | 1 | |a Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila |
| 264 | 1 | |c 2013 | |
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| 502 | |a Würzburg, Univ., Diss., 2013 | ||
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Datensatz im Suchindex
| _version_ | 1804151786886397952 |
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| adam_text | 2 TABLE OF CONTENTS
TABLE OF CONTENTS
SUMMARY 6
ZUSAMMENFASSUNG
7
LYRICAL ABSTRACT 8
INTRODUCTION 9
1. CHEMICAL SYNAPSES U
1.1. SELECTED PROTEINS AT THE PRESYNAPTIC ACTIVE ZONE 12
1.1.1. BRUCHPILOT/ELKS, BASSOON AND PICCOLO 12
1.1.2. RIM, RIM-BINDING PROTEIN AND MUNCI3 13
1.1.3. LIPRIN-(X AND SYD-I 14
1.2. RECEPTORS AT THE POSTSYNAPTIC DENSITY 14
1.2.1. IONOTROPIC NEUROTRANSMITTER RECEPTORS 15
1.2.2. METABOTROPIC RECEPTORS 16
2. SYNAPTIC PLASTICITY 17
2.1. SHORT-TERM PLASTICITY 17
2.2. LONG-TERM PLASTICITY 17
2.2.1. LONG-TERM POTENTIATION 17
2.2.2. ADDITIONAL MECHANISMS FOR LONG-TERM PLASTICITY 18
2.2.2.1. CASPASES AND LONG-TERM PLASTICITY 18
2.2.2.2. LOCAL SYNAPTIC TRANSLATION 19
3. ANATOMY OF THE DROSOPHILA CNS
20
3.1. THE OLFACTORY PATHWAY 21
3.1.1. THE ANTENNAL LOBES 21
3.1.2. THE MUSHROOM BODIES 22
3.2. THE CENTRAL COMPLEX 24
3.3. GENETIC TOOLS FOR THE ANALYSIS OF THE
DROSOPHILA
CNS 25
4. SELECTED ADAPTIVE BEHAVIOURS OF DROSOPHILA 26
4.1. AVERSIVE OLFACTORY LEARNING AND MEMORY 26
4.2. COURTSHIP CONDITIONING 28
4.3. ETHANOL-MEDIATED BEHAVIOURS 29
MATERIALS & METHODS
31
1.
ANIMAL REARING AND FLY STRAINS
32
1.1. FLY STRAINS GENERATED IN THE CONTEXT OF THIS THESIS 32
1.2. PREVIOUSLY PUBLISHED FLY STRAINS USED IN THIS THESIS 33
2. IMAGING 33
2.1. IMMUNOHISTOCHEMISTRY 33
2.1.1. SOLUTIONS ; 33
2.1.2. ANTIBODIES 34
2.1.3. STAINING PROTOCOLS 34
HTTP://D-NB.INFO/1047491176
TABLE OF CONTENTS 3
2.1.4. IMAGE ACQUISITION 35
2.2.
IN VIVO
IMAGING 35
2.3. IMAGE POST-PROCESSING AND ANALYSIS 35
2.4. FUNCTIONAL CALCIUM IMAGING 36
2.5. IMMUNOELECTRON MICROSCOPY 36
3.
RATIOMETRIC ANALYSIS OF PRESYNAPTIC PROTEINS
37
3.1. SEGMENTATION AND THRESHOLDING 37
3.2. CALCULATION OF RATIO VALUES 40
3.3. ANALYSIS OF RATIOMETRIC RESULTS AND STATISTICS 42
3.4. ADDITIONAL MODULES 44
4. MOLECULAR CLONING
45
4.1. CONSTRUCTS FOR THE GENERATION OF TRANSGENIC FLIES 45
4.1.1. UAS CONSTRUCTS 45
4.1.2. P[ACMAN] CONSTRUCTS 46
4.1.3. MISCELLANEOUS CONSTRUCTS 47
4.2. ANTIBODIES 48
4.3. SINGLE-FLY PCR 48
4.4. IN SITU
HYBRIDIZATION 49
5.
BIOCHEMISTRY
49
5.1. ETHANOL ABSORPTION 49
5.2.
IN VITRO
EXPERIMENTS USING PURIFIED DREP-2 AND -3 49
5.2.1. PROTEIN EXPRESSION AND PROTEIN PURIFICATION 49
5.2.2. NUCLEASE ACTIVITY ASSAY 50
5.2.3. CASPASE CLEAVAGE ASSAYS 50
5.3. PULLDOWN EXPERIMENTS 50
5.4. FLY HEAD EXTRACTS FOR WESTERN BLOTTING 50
5.5. WESTERN BLOTTING 51
5.6. SYNAPTOSOME PREPARATIONS 51
5.7. QUANTITATIVE MASS SPECTROMETRY
;
51
5.7.1. QUANTITATIVE AFFINITY PURIFICATION AND MASS SPECTROMETRY 51
5.7.2. PULLDOWN EXPERIMENTS 52
5.7.3. LIQUID CHROMATOGRAPHY MS/MS ANALYSIS 52
5.7.4. GENERATION OF THE NETWORK OF INTERACTORS 53
6. BEHAVIOURAL ANALYSES
54
6.1. OLFACTORY CONDITIONING 54
6.2. COURTSHIP CONDITIONING 54
6.3. ETHANOL SEDATION ^ 55
6.4. LOCOMOTOR ACTIVITY 55
7.
MISCELLANEOUS METHODS 56
7.1. ELECTROPHYSIOLOGY 56
7.2. ADULT LIFE SPAN 56
4 TABLE OF CONTENTS
7.3. GENERAL DATA ANALYSIS AND STATISTICS 56
RESULTS 57
1. CHARACTERIZATION OF SYNAPTIC DIVERSITY USING RATIOMETRIC ANALYSES 58
1.1. COMPARISON OF EXPRESSION PATTERNS OF PRESYNAPTIC PROTEINS 58
1.2. A RATIOMETRIC METHOD FOR THE DESCRIPTION OF DIFFERENCES IN CAZ
PROTEIN
EXPRESSION 63
1.2.1. SEGMENTATION AND THRESHOLDING 64
1.2.2. CALCULATION OF RATIOS 65
1.3. CHARACTERIZATION OF SYNAPSE TYPES 66
1.3.1. MUSHROOM BODY CALYX 66
1.3.2. ANTENNALLOBE 74
1.4. CHANGES IN THE CAZ PROTEIN COMPOSITION 81
1.4.1. ANALYSIS OF THE REDUCTION OF BRP LEVELS IN KENYON CELLS 81
1.4.2. ALTERED RATIO CLASSES IN THE SHAKB
3
MUTANT 82
1.4.3. DETECTING CHANGES IN PRESYNAPSE TYPE ABUNDANCE AFTER CLIPPING
ANTENNAE 85
2. NOVEL SYNAPTIC PROTEINS: THE DREP FAMILY 87
2.1. CHARACTERIZATION OF DREP-2 87
2.1.1. DISCOVERY OF DREP-2 AS A NOVEL SYNAPTIC PROTEIN 87
2.1.2. GENERATION OF DREP-2 ANTIBODIES AND DREP-2 MUTANTS 88
2.1.3. DREP-2 CO-PRECIPITATES WITH BRUCHPILOT 89
2.1.4. DREP-2 AT THE LARVAL NEUROMUSCULAR JUNCTION 90
2.1.5. DREP-2 IS TRANSPORTED BIDIRECTIONALLY ALONG AXONS 92
2.1.6. LOCALIZATION OF DREP-2 IN THE CENTRAL NERVOUS SYSTEM 94
2.1.6.1. DREP-2 LOCALIZES TO POSTSYNAPTIC DENSITIES AT MB CALYX
MICROGLOMERULI 96
2.1.6.2. EVIDENCE FOR DREP-2 AT RECURRENT KC SYNAPSES IN THE CALYX 101
2.1.7. DREP-2 EXPRESSION IN RELATION TO THE EXPRESSION OF
NEUROTRANSMITTERS 102
2.1.7.1. COLOCALIZATION OF DREP-2 WITH ACETYLCHOLINE RECEPTORS 102
2.1.7.2. EXPRESSION PATTERNS OF DIFFERENT NEUROTRANSMITTER RECEPTORS IN
THE CALYX,
RELATIVE TO DREP-2 103
2.1.7.3. EXAMINATION OF WHETHER DREP-2 CAN DIRECTLY INTERACT WITH
METABOTROPIC
GLUTAMATE RECEPTORS 105
2.1.8. CHARACTERIZATION OF THE DREP-2 MUTANT 105
2.1.8.1. THE OVERALL MORPHOLOGY OF MUTANT BRAINS APPEARS LARGELY NORMAL
105
2.1.8.2. APOPTOSIS DOES NOT APPEAR TO BE MISREGULATED IN DREP-2 MUTANTS
106
2.1.8.3. DREP-2 MUTANTS LIVE SHORTER THAN WILDTYPE FLIES 107
2.1.8.4. DREP-2 IS NOT REQUIRED FOR BASIC TRANSMISSION IN THE VISUAL
SYSTEM 108
2.1.8.5. DREP-2 IS REQUIRED FOR NORMAL OLFACTORY LEARNING 109
2.1.8.6. LOWERED ODOUR-EVOKED CA
2+
RESPONSES IN KCS OF MUTANTS 112
2.1.8.7. ETHANOL HYPERSENSITIVITY OF DREP-2 MUTANTS 113
2.1.8.8. LOCOMOTOR HYPERACTIVITY OF DREP-2 MUTANTS 117
2.1.9. MASS SPECTROMETRY: DREP-2 IN COMPLEXES WITH TRANSLATIONAL
REPRESSORS 118
2.1.9.1. MEMBRANE PROTEINS 123
2.1.9.2. RNA-BINDING PROTEINS 123
TABLE OF CONTENTS 5
2.1.9.3. DREP-2 ANTAGONIZES FMRP IN COURTSHIP CONDITIONING 124
2.2. A BRIEF GLANCE AT OTHER DREP FAMILY MEMBERS 127
2.2.1. DREP-4 127
2.2.2. DREP-I 128
2.2.3. DREP-3, A PUTATIVE INTERACTOR OF DREP-2 128
2.2.3.1. THE LOCALIZATION OF DREP-3
GFP
IS HIGHLY SIMILAR TO DREP-2 130
2.2.3.2. PUTATIVE CLEAVAGE OF DREP-3 BY CASPASES 131
DISCUSSION 133
1. CHARACTERIZATION OF SYNAPTIC DIVERSITY USING RATIOMETRIC ANALYSES 134
1.1. ADVANTAGES OF THE RATIOMETRIC METHOD 134
1.1.1. COMPARISON TO AN ANALYSIS OF ABSOLUTE INTENSITY VALUES 134
1.1.2. ANALYSIS OF IMAGE STACKS 135
1.2. TOWARDS A MOLECULAR ATLAS OF SYNAPSE TYPES 136
1.2.1. MIXED SYNAPSE POPULATIONS 136
1.2.1.1. ANALYSIS OF THE SHAKB
2
MUTANT 137
1.2.1.2. ANALYSIS OF FLIES LACKING ANTENNAE 138
1.2.2. CURRENT LEVEL OF COVERAGE OF SYNAPSE TYPES 138
1.3. APPLICATIONS OF THE METHOD 138
1.3.1. THE RATIONAL APPROACH TOWARDS A MEANINGFUL CONNECTOME 140
2. THE ROLE OF DREP-2 AT SYNAPSES 141
2.1. WHAT IS THE FUNCTION OF DREP-2 IN KENYON CELLS? 142
2.1.1. PUTATIVE ROLES FOR DREP-2 AND G-PROTEIN SIGNALLING IN THE CALYX
143
2.1.2. DREP-2 COULD REGULATE METABOTROPIC SIGNALLING IN GENERAL 144
2.1.3. DOES DREP-2 INDIRECTLY REGULATE CAMP LEVELS? 144
2.1.4. DREP-2 AT MUSHROOM BODY LOBES 146
2.2. PRESYNAPTIC DREP-2 146
2.3. CONNECTIONS OF DREP-2 TO THE FRAGILE X PROTEIN 148
2.3.1. INTERFERENCE WITH MGLUR-MEDIATED PROTEIN SYNTHESIS 150
2.3.2. A ROLE FOR DREP-2 IN BALANCING MGLUR SIGNALLING? 150
2.4. A DIRECT CONTRIBUTION OF DREP-2 TO TRANSLATIONAL REGULATION? 152
2.4.1. CASPASE-MEDIATED PLASTICITY 153
APPENDIX 155
REFERENCES 156
INDEX OF FIGURES 176
INDEX OF TABLES 179
INDEX OF ABBREVIATIONS 180
ACKNOWLEDGEMENTS 182
LIST OF PUBLICATIONS 184
CURRICULUM VITAE
:
185
THIS THESIS CAN BE DOWNLOADED IN ELECTRONIC FORM AT
HTTP://BIOLOGIE.ANDLAUER.NET/THESIS.PDF
|
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| spelling | Andlauer, Till 1979- Verfasser (DE-588)138094020 aut Structural and functional diversity of synapses in the drosophila CNS = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila von Till Felix Malte Andlauer Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila 2013 186 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. und engl. Sprache Würzburg, Univ., Diss., 2013 Neurobiologie (DE-588)4041871-6 gnd rswk-swf Synapse (DE-588)4184224-8 gnd rswk-swf Zentralnervensystem (DE-588)4067637-7 gnd rswk-swf Taufliege (DE-588)4078168-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Taufliege (DE-588)4078168-9 s Neurobiologie (DE-588)4041871-6 s Synapse (DE-588)4184224-8 s Zentralnervensystem (DE-588)4067637-7 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-85018 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-85018 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=027037910&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Andlauer, Till 1979- Structural and functional diversity of synapses in the drosophila CNS = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila Neurobiologie (DE-588)4041871-6 gnd Synapse (DE-588)4184224-8 gnd Zentralnervensystem (DE-588)4067637-7 gnd Taufliege (DE-588)4078168-9 gnd |
| subject_GND | (DE-588)4041871-6 (DE-588)4184224-8 (DE-588)4067637-7 (DE-588)4078168-9 (DE-588)4113937-9 |
| title | Structural and functional diversity of synapses in the drosophila CNS = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila |
| title_alt | Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila |
| title_auth | Structural and functional diversity of synapses in the drosophila CNS = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila |
| title_exact_search | Structural and functional diversity of synapses in the drosophila CNS = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila |
| title_full | Structural and functional diversity of synapses in the drosophila CNS = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila von Till Felix Malte Andlauer |
| title_fullStr | Structural and functional diversity of synapses in the drosophila CNS = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila von Till Felix Malte Andlauer |
| title_full_unstemmed | Structural and functional diversity of synapses in the drosophila CNS = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila von Till Felix Malte Andlauer |
| title_short | Structural and functional diversity of synapses in the drosophila CNS |
| title_sort | structural and functional diversity of synapses in the drosophila cns strukturelle und funktionale diversitat von synapsen im zns von drosophila |
| title_sub | = Strukturelle und funktionale Diversität von Synapsen im ZNS von Drosophila |
| topic | Neurobiologie (DE-588)4041871-6 gnd Synapse (DE-588)4184224-8 gnd Zentralnervensystem (DE-588)4067637-7 gnd Taufliege (DE-588)4078168-9 gnd |
| topic_facet | Neurobiologie Synapse Zentralnervensystem Taufliege Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-85018 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027037910&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT andlauertill structuralandfunctionaldiversityofsynapsesinthedrosophilacnsstrukturelleundfunktionalediversitatvonsynapsenimznsvondrosophila AT andlauertill strukturelleundfunktionalediversitatvonsynapsenimznsvondrosophila |