The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease: = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Erlangen ; Nürnberg
[2017]
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Physical Description: | III, 140 Seiten Illustrationen, Diagramme 21 cm |
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MARC
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| 100 | 1 | |a Menges, Stefanie |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease |b = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom |c vorgelegt von Stefanie Menges |
| 246 | 1 | 3 | |a Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom |
| 246 | 1 | 1 | |a Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom |
| 264 | 1 | |a Erlangen ; Nürnberg |c [2017] | |
| 300 | |a III, 140 Seiten |b Illustrationen, Diagramme |c 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
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| 502 | |b Dissertation |c Friedrich-Alexander-Universität Erlangen-Nürnberg |d 2017 | ||
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Record in the Search Index
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|---|---|
| adam_text | CONTENTS
LIST OF FIGURES..
DECLARATION...................................................................................................................................4
SUMMARY......................................................................................................................................5
ZUSAMMENFASSUNG......................................................................................................................
7
1
INTRODUCTION..........................................................................................................................10
1.1 ALPHA-SYNUCLEIN IN PARKINSON S
DISEASE.........................................................................
10
1.1.1 DISCOVERY OF
ASYN............................................................................................
10
1.1.2 CLINICAL AND PATHOLOGICAL ASPECTS OF PD
..........................................................
11
1.1.3 PD-RELATED ASYN
MUTATIONS...............................................................................12
1.1.4 STRUCTURE AND FUNCTION OF
ASYN.........................................................................14
1.1.5 ASYN
AGGREGATION..............................................................................................
16
1.2 OXIDATIVE
STRESS..............................................................................................................17
1.2.1 DEFINITION AND FORMATION OF OXIDATIVE STRESS
...................................................
17
1.2.2 OXIDATIVE STRESS IN PD
......................................................................................19
1.2.3 ASYN AND OXIDATIVE
STRESS................................................................................
20
1.3 MITOCHONDRIAL
BIOLOGY...................................................................................................
22
1.3.1 MITOCHONDRIAL
FUNCTION.....................................................................................22
1.3.2 MITOCHONDRIAL
MORPHOLOGY.............................................................................
24
1.3.3 MITOCHONDRIAL QUALITY CONTROL
PATHWAYS.........................................................26
1.3.4 MITOCHONDRIAL DYSFUNCTION IN PD
....................................................................29
1.3.5 ASYN AND MITOCHONDRIAL
DYSFUNCTION...............................................................
29
2 HYPOTHESES AND
AIMS..........................................................................................................31
3 MATERIAL AND
METHODS.........................................................................................................
33
3.1
MATERIALS.........................................................................................................................33
3.1.1
CHEMICALS.........................................................................................................33
3.1.2 SPECIFIC REAGENTS AND ENZYMES
........................................................................
33
3.1.3 KITS AND MASTER MIXES
.....................................................................................
34
3.1.4 BUFFERS AND
SOLUTIONS........................................................................................35
3.1.5 DNA AND PROTEIN STANDARDS
.............................................................................
37
3.1.6
PLASMIDS...........................................................................................................
37
3.1.7
ANTIBODIES........................................................................................................38
3.1.7.1 PRIMARY ANTIBODIES
............................................................................
38
3.1.7.2 SECONDARY ANTIBODIES
........................................................................
39
3.1.8 MEDIA AND REAGENTS FOR CELL CULTURE
.................................................................
39
~ I ~
3.1.9 STABLE CELL
LINES.................................................................................................
41
3.1.10 BACTERIAL STRAINS AND M
EDIA..............................................................................41
3.1.11 SPECIFIC
CONSUMABLES.......................................................................................
41
3.1.12 TECHNICAL
DEVICES.............................................................................................
42
3.1.13
SOFTWARE............................................................................................................43
3.2
METHODS........................................................................................................................44
3.2.1 CELL
CULTURE.......................................................................................................
44
3.2.1.1 H4 AND LUHMES
CELLS.....................................................................44
3.2.1.2 HUMAN FIBROBLASTS, HUMAN IPSC-DERIVED NPCS AND DIFFERENTIATED
NEURAL
CULTURES....................................................................................
45
3.2.1.3 RAT PRIMARY CORTICAL NEURON CULTURES
................................................
45
3.2.2 CELL BIOLOGICAL
METHODS....................................................................................
46
3.2.2.1 CALCIUM-PHOSPHATE TRANSFECTION
.......................................................
46
3.2.2.2 CELL
TREATMENTS...................................................................................
46
3.2.2.3 MITOTRACKER AND LYSOTRACKER STAININGS
...........................................
46
3.2.2.4 CELL VIABILITY ASSAYS AND ATP LEVELS
...............................................
47
3.2.2.5 FLOW
CYTOMETRY..................................................................................47
3.2.2.6
IMMUNOCYTOCHEMISTRY.......................................................................49
3.2.2.7 MORPHOLOGICAL ANALYSIS AND QUANTIFICATION OF MITOSPHERES
.............
49
3.2.2.8 HIGH-RESOLUTION RESPIROMETRY
...........................................................
49
3.2.3 MOLECULAR BIOLOGICAL AND BIOCHEMICAL METHODS
..............................................
50
3.2.3.1 GENERATION OF SHRNA
VECTORS............................................................50
3.2.3.2 SITE-DIRECTED
MUTAGENESIS..................................................................52
3.2.3.3 TRANSFORMATION OF COMPETENT CELLS
...................................................
53
3.2.3.4 PLASMID ISOLATION (MINIPREP AND MAXIPREP)
....................................
53
3.2.3.5 PREPARATION OF RECOMBINANT HUMAN ASYN
..........................................
53
3.2.3.6 INDUCTION OF POST-TRANSLATIONAL MODIFICATIONS
...................................
55
3.2.3.7 MASS
SPECTROMETRY.............................................................................55
3.2.3.8 IN VITRO FIBRILLIZATION OF ASYN AND THIOFLAVIN-T
ASSAY......................56
3.2.3.9 SIZE EXCLUSION CHROMATOGRAPHY
........................................................
56
3.2.3.10 SUCROSE DENSITY GRADIENT CENTRIFUGATION
.........................................
57
3.2.3.11 SOLUBILITY
ASSAY................................................................................
57
3.2.3.12 ELECTRON
MICROSCOPY........................................................................58
3.2.3.13 PREPARATION OF A CRUDE MITOCHONDRIAL FRACTION
.................................
58
3.2.3.14 SDS-PAGE AND WESTERN BLOTTING
...................................................
58
3.2.3.15
OXYBLOT............................................................................................
59
3.2.4
STATISTICS...........................................................................................................
60
4
RESULTS....................................................................................................................................61
4.1 THE ROLE OF H50 ALTERATIONS IN OXIDATIVE STRESS-INDUCED ASYN
PATHOLOGY
.................
61
4.1.1 H50 IS THE MOST REACTIVE RESIDUE FOR HNE-MODIFICATION OF ASYN
.................
61
4.1.2 H50 IS CRUCIAL FOR HNE-MEDIATED OLIGOMERIZATION OF ASYN AND
CYTOTOXICITY 63
4.1.3 H50Q MUTATION INCREASES THE AGGREGATION PROPENSITY OF ASYN BOTH IN
VITRO
AND IN
CELLS........................................................................................................
67
4.1.4 OVEREXPRESSION OF H50Q ASYN INCREASES CELLULAR SUSCEPTIBILITY TO
OXIDATIVE
STRESS.................................................................................................................
70
4.2 THE EFFECT OF ASYN ON MITOCHONDRIAL HOMEOSTASIS UNDER OXIDATIVE
STRESS
................
72
4.2.1 H202-INDUCED OXIDATIVE STRESS TRIGGERS THE FORMATION OF
MITOSPHERES
...........
72
4.2.2 MITOSPHERE FORMATION IS A COMMON CELLULAR RESPONSE TO OXIDATIVE
STRESS
.....
77
4.2.3 THE INVOLVEMENT OF FISSION AND FUSION IN MITOSPHERE FORMATION
..................
79
4.2.4 MITOSPHERES ARE NOT LINKED TO PARKIN-ASSOCIATED
MITOPHAGY..........................81
4.2.5 MITOSPHERE FORMATION IS LINKED TO CASPASE3 ACTIVATION AND
APOPTOSIS
.........
84
4.2.6 ASYN PREVENTS OXIDATIVE STRESS-RELATED MITOSPHERE
FORMATION.........................84
4.2.7 ASYN INFLUENCES FUSION OF MITOCHONDRIA UNDER BASAL CONDITIONS
...................
90
4.2.8 ASYN PREVENTS CASPASE3 ACTIVATION AND ATTENUATES THE IMPAIRMENT OF
MITOCHONDRIAL FUNCTION UNDER OXIDATIVE
STRESS................................................92
4.2.9 MITOCHONDRIAL ALTERATIONS UNDER 6-OHDA AND
ROTENONE...............................94
4.2.10 ASYN PROTECTS AGAINST 6-OHDA-INDUCED MITOSPHERES AND APOPTOSIS,
BUT NOT
ROTENONE-INDUCED MITOCHONDRIAL ALTERATIONS AND CYTOTOXICITY
.......................
95
5
DISCUSSION.............................................................................................................................
98
5.1 THE RELEVANCE OF H50 ALTERATIONS IN PD-ASSOCIATED ASYN PATHOLOGY
.........................
98
5.1.1 THE ROLE OF H50 IN HNE-MEDIATED ASYN
PATHOLOGY........................................98
5.1.2 THE IMPACT OF H50Q MUTATION ON ASYN AGGREGATION AND TOXICITY
.................
100
5.2 THE ROLE OF ASYN IN MITOCHONDRIAL HOMEOSTASIS UNDER OXIDATIVE STRESS
....................
106
5.2.1 *MITOSPHERES* - A DISTINCT MITOCHONDRIAL PHENOTYPE UNDER OXIDATIVE
STRESS 106
5.2.2 THE FUNCTION OF ASYN IN MITOCHONDRIAL RESPONSE TO OXIDATIVE
STRESS.............110
5.3
CONCLUSIONS................................................................................................................
118
BIBLIOGRAPHY...........................................................................................................................
120
ABBREVIATIONS..........................................................................................................................
132
ACKNOWLEDGEMENT..................................................................................................................
135
SUPPLEMENTS............................................................................................................................
137
|
| any_adam_object | 1 |
| author | Menges, Stefanie |
| author_facet | Menges, Stefanie |
| author_role | aut |
| author_sort | Menges, Stefanie |
| author_variant | s m sm |
| building | Verbundindex |
| bvnumber | BV044862193 |
| ctrlnum | (OCoLC)1136332259 (DE-599)DNB1152101153 |
| discipline | Biologie Medizin |
| format | Thesis Book |
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| id | DE-604.BV044862193 |
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| indexdate | 2024-07-10T08:03:09Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-030256847 |
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| physical | III, 140 Seiten Illustrationen, Diagramme 21 cm |
| publishDate | 2017 |
| publishDateSearch | 2017 |
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| spelling | Menges, Stefanie Verfasser aut The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom vorgelegt von Stefanie Menges Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom Erlangen ; Nürnberg [2017] III, 140 Seiten Illustrationen, Diagramme 21 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Friedrich-Alexander-Universität Erlangen-Nürnberg 2017 Parkinson-Krankheit (DE-588)4044683-9 gnd rswk-swf Oxidativer Stress (DE-588)4353497-1 gnd rswk-swf Mitochondriopathie (DE-588)4280150-3 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Parkinson-Krankheit (DE-588)4044683-9 s Oxidativer Stress (DE-588)4353497-1 s Mitochondriopathie (DE-588)4280150-3 s DE-604 B:DE-101 application/pdf http://d-nb.info/1152101153/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030256847&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Menges, Stefanie The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom Parkinson-Krankheit (DE-588)4044683-9 gnd Oxidativer Stress (DE-588)4353497-1 gnd Mitochondriopathie (DE-588)4280150-3 gnd |
| subject_GND | (DE-588)4044683-9 (DE-588)4353497-1 (DE-588)4280150-3 (DE-588)4113937-9 |
| title | The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom |
| title_alt | Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom |
| title_auth | The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom |
| title_exact_search | The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom |
| title_full | The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom vorgelegt von Stefanie Menges |
| title_fullStr | The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom vorgelegt von Stefanie Menges |
| title_full_unstemmed | The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom vorgelegt von Stefanie Menges |
| title_short | The interplay of alpha-synuclein, oxidative stress and mitochondrial dysfunction in Parkinson's disease |
| title_sort | the interplay of alpha synuclein oxidative stress and mitochondrial dysfunction in parkinson s disease das zusammenspiel von alpha synuklein oxidativem stress und mitochondrialer dysfunktion beim parkinson syndrom |
| title_sub | = Das Zusammenspiel von alpha-Synuklein, oxidativem Stress und mitochondrialer Dysfunktion beim Parkinson-Syndrom |
| topic | Parkinson-Krankheit (DE-588)4044683-9 gnd Oxidativer Stress (DE-588)4353497-1 gnd Mitochondriopathie (DE-588)4280150-3 gnd |
| topic_facet | Parkinson-Krankheit Oxidativer Stress Mitochondriopathie Hochschulschrift |
| url | http://d-nb.info/1152101153/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030256847&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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