Verfügbarkeit: The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease

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Titel:	The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease = Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom
Weiterer Titel:	Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom
Von:	vorgelegt von Georgia Minakaki
Person:	Minakaki, Georgia Verfasser aut
Hauptverfasser:	Minakaki, Georgia (VerfasserIn)
Format:	Abschlussarbeit Buch
Sprache:	Englisch Deutsch
Veröffentlicht:	Erlangen ; Nürnberg [2018]
Schlagworte:	Exosom > Vesikel Autophagie > Physiologie Parkinson-Krankheit Synuclein > alpha- Hochschulschrift
Online-Zugang:	http://d-nb.info/1166464741/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030591137&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
Beschreibung:	Zusammenfassung in deutscher Sprache
Beschreibung:	VI, 124 Seiten Illustrationen, Diagramme 21 cm

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245	1	0	\|a The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease \|b = Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom \|c vorgelegt von Georgia Minakaki
246	1	3	\|a Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom
246	1	1	\|a Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom
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Datensatz im Suchindex

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adam_text	CONTENTS LIST OF FIGURES AND TABLES.................................................................................................... IV DECLARATION............................................................................................................................. VI SUMMARY...................................................................................................................................1 ZUSAMMENFASSUNG................................................................................................................... 3 1 INTRODUCTION........................................................................................................................ 6 1.1 ALPHA-SYNUCLEIN IN SYNUCLEINOPATHIES.........................................................................6 1.1.1 CLINICAL SYMPTOMS, PREVALENCE AND TREATMENT OF PARKINSON S DISEASE ... 6 1.1.2 FROM LEWY BODIES TO THE DEFINITION OF SYNUCLEINOPATHIES ...................... 7 1.1.3 ASYN STRUCTURE AND PROPOSED PHYSIOLOGICAL FUNCTION..............................9 1.2 CELLULAR PATHWAYS IMPLICATED IN PARKINSON S DISEASE ................................................ 11 1.2.1 GENETIC BACKGROUND OF PARKINSON S DISEASE ......................................... 11 1.2.2 UBIQUITIN-PROTEASOME AND LYSOSOMAL DEFECTS.......................................12 1.2.3 THE MITOCHONDRIAL DAMAGE REPAIR PATHWAY..........................................14 1.3 DEGRADATION OF ASYN VIA THE AUTOPHAGY-LYSOSOME PATHWAY (ALP) ........................... 16 1.3.1 ALP INHIBITION IN SYNUCLEINOPATHIES ..................................................... 16 1.4 HIERARCHICAL SPREADING OF LEWY BODY PATHOLOGY....................................................... 18 1.4.1 ASYN RELEASE AND TRANSFER IN SYNUCLEINOPATHY PROGRESSION ................... 20 1.4.2 INTRACELLULAR PROCESSING AND ASYN RELEASE PATHWAYS CROSS-TALK............21 1.4.3 ALP INHIBITION INCREASES ASYN RELEASE, PARALLELED BY TOXIC MICROENVIRONMENTAL RESPONSES .................................................................................. 23 2 HYPOTHESES AND AIMS........................................................................................................ 24 2.1 THE ROLE OF LYSOSOMAL INHIBITION IN SYNUCLEINOPATHY PROGRESSION ............................. 24 2.1.1 HYPOTHESIS 1: INHIBITION OF ALP FLUX STIMULATES ASYN RELEASE VIA EVS. 25 2.1.2 HYPOTHESIS 2: ALP INHIBITION HAS IMPLICATIONS FOR EV BIOGENESIS AND GENERAL BIOCHEMICAL PROFILE ....................................................................................... 25 2.1.3 HYPOTHESIS 3: EVS PLAY A ROLE IN THE PROGRESSION OF SYNUCLEINOPATHY. 26 2.2 ALP MODULATION AS A THERAPEUTIC TARGET IN PD...........................................................26 3 MATERIALS AND METHODS.................................................................................................... 27 3.1 MATERIALS..................................................................................................................27 3.1.1 CHEMICALS............................................................................................. 27 3.1.2 SPECIFIC REAGENTS AND ENZYMES.............................................................27 3.1.3 KITS....................................................................................................... 28 3.1.4 BUFFERS AND SOLUTIONS ........................................................................... 28 I 3.1.5 PROTEIN MW REFERENCE........................................................................... 30 3.1.6 ANTIBODIES............................................................................................. 30 3.1.7 CELL CULTURE MEDIA AND REAGENTS............................................................31 3.1.8 CONSUMABLES......................................................................................... 32 3.1.9 TECHNICAL DEVICES.................................................................................. 33 3.1.10 SOFTWARE................................................................................................ 35 3.2 METHODS FOR HYPOTHESES 1-3...................................................................................... 35 3.2.1 CELL CULTURE............................................................................................ 35 3.2.2 TREATMENTS AND CELLULAR ANALYSES.......................................................... 37 3.2.3 PREPARATION OF EV FRACTIONS.................................................................. 40 3.2.4 EV PROTEOMICS...................................................................................... 43 3.2.5 INTRA-CORTICAL INJECTION OF CSF-DERIVED EVS INTO MICE ......................... 44 3.2.6 SUBJECTS................................................................................................. 44 3.2.7 IMMUNOHISTOCHEMICAL ANALYSIS OF IN VIVO ASYN TRANSFER ...................... 45 3.3 EXPERIMENTAL PROCEDURES FOR HYPOTHESIS 4 ............................................................... 45 3.3.1 MICE...................................................................................................... 45 3.3.2 BEHAVIORAL PROCEDURES.......................................................................... 46 3.3.3 ANTIBODIES............................................................................................. 47 3.3.4 WESTERN BLOT ANALYSES........................................................................... 47 3.3.5 IMMUNOHISTOCHEMISTRY......................................................................... 48 3.4 STATISTICAL ANALYSES..................................................................................................... 48 4 RESULTS............................................................................................................................... 49 4.1 THE ROLE OF LYSOSOMAL INHIBITION IN SYNUCLEINOPATHY PROGRESSION............................49 4.1.1 LYSOSOMAL INHIBITION ENHANCES ASYN RELEASE ........................................ 49 4.1.2 ASYN IS RELEASED VIA EVS...................................................................... 52 4.1.3 EVS RELEASED UNDER ALP INHIBITION CONTAIN HIGHER ASYN CARGO .......... 56 4.1.4 THE EFFECT OF ALP MODULATORS AND ROTENONE ON AUTOPHAGOSOMAL FLUX 58 4.1.5 A WIDESPREAD, FUSED MVB-AP COMPARTMENT IS OBSERVED UNDER ALP INHIBITION 60 4.1.6 LC3-II AND P62 ARE PROTEIN COMPONENTS OF EVS AND BECOME ENRICHED DURING ALP INHIBITION................................................................................................ 61 4.1.7 HUMAN CSF EVS ARE LC3-II AND P62 POSITIVE .................................... 65 4.1.8 CSF EVS ARE INVOLVED IN THE CELL-TO-CELL TRANSFER OF ASYN IN VIVO ........ 68 4.2 VALIDATION OF THE ALP AS AN INTERVENTIONAL TARGET FOR PD......................................... 73 4.2.1 ASYN MOUSE MODELS AND EXPERIMENTAL SETUP ....................................... 73 4.2.2 TREADMILL EXERCISE INTERVENTION DID NOT ROBUSTLY INDUCE ALP FLUX ....... 75 4.2.3 TREADMILL EXERCISE INTERVENTION MODERATELY INFLUENCED HUWT/KO LIVER MITOCHONDRIAL HOMEOSTASIS ....................................................................................... 78 4.2.4 CORTICAL AND STRIATAL ASYN LEVELS WERE NOT AFFECTED BY TREADMILL EXERCISE INTERVENTION 79 4.2.5 GAIT DEFICITS AND POSTURAL INSTABILITY OBSERVED IN ASYN MOUSE MODELS AT BASELINE 80 4.2.6 TREADMILL EXERCISE INTERVENTION SIGNIFICANTLY IMPROVED GAIT AND POSTURAL INSTABILITY IN ASYN MOUSE MODELS...............................................................................82 5 DISCUSSION..........................................................................................................................90 5.1 LYSOSOMAL INHIBITION IS LINKED TO SYNUCLEINOPATHY PROGRESSION.............................90 5.1.1 LYSOSOMAL INHIBITION INCREASES ASYN CARGO WITHIN EVS ...................... 90 5.1.2 ALP AND EV RELEASE ARE CONVERGING MECHANISMS GOVERNING ASYN HOMEOSTASIS IN THE CNS ............................................................................................ 91 5.1.3 AUTOPHAGO-EXOSOMES; EVIDENCE FOR AN EXISTING ALP-MVB-EV ROUTE. 92 5.1.4 EVS PLAY AN IMPORTANT ROLE IN THE CELL-TO-CELL TRANSFER OF ASYN IN VIVO 94 5.2 PHYSICAL EXERCISE HAS THERAPEUTIC POTENTIAL FOR PD ...................................................95 5.2.1 TREADMILL EXERCISE INTERVENTION AS AN INDUCER OF THE ALP .................... 95 5.2.2 ASYN MOUSE MODELS DISPLAY GAIT AND POSTURAL CONTROL DEFICITS ............... 96 5.2.3 TREADMILL EXERCISE INTERVENTION SIGNIFICANTLY IMPROVES GAIT AND POSTURAL CONTROL DEFICITS........................................................................................................... 97 5.3 CONCLUSIONS............................................................................................................... 99 6 BIBLIOGRAPHY.....................................................................................................................101 7 ABBREVIATIONS...................................................................................................................119 8 ACKNOWLEDGMENTS............................................................................................................122
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spellingShingle	Minakaki, Georgia The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease = Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom Exosom Vesikel (DE-588)1154814491 gnd Autophagie Physiologie (DE-588)4441664-7 gnd Parkinson-Krankheit (DE-588)4044683-9 gnd Synuclein alpha- (DE-588)1156563224 gnd
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title	The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease = Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom
title_alt	Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom
title_auth	The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease = Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom
title_exact_search	The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease = Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom
title_full	The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease = Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom vorgelegt von Georgia Minakaki
title_fullStr	The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease = Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom vorgelegt von Georgia Minakaki
title_full_unstemmed	The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease = Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom vorgelegt von Georgia Minakaki
title_short	The functional link between autophagy, exosomes and the transcellular spread of alpha-synuclein in Parkinson's disease
title_sort	the functional link between autophagy exosomes and the transcellular spread of alpha synuclein in parkinson s disease die funktionelle verbindung zwischen autophagie exosomen und der transzellularen ausbreitung von alpha synuklein beim parkinson syndrom
title_sub	= Die funktionelle Verbindung zwischen Autophagie, Exosomen und der transzellulären Ausbreitung von Alpha-Synuklein beim Parkinson-Syndrom
topic	Exosom Vesikel (DE-588)1154814491 gnd Autophagie Physiologie (DE-588)4441664-7 gnd Parkinson-Krankheit (DE-588)4044683-9 gnd Synuclein alpha- (DE-588)1156563224 gnd
topic_facet	Exosom Vesikel Autophagie Physiologie Parkinson-Krankheit Synuclein alpha- Hochschulschrift
url	http://d-nb.info/1166464741/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030591137&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
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