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Mechanism of Extracellular Aggregate-induced Alpha-synuclein Transmission
DC Field | Value | Language |
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dc.contributor.advisor | 이승재 | - |
dc.contributor.author | 임윤주 | - |
dc.date.accessioned | 2018-05-29T04:51:18Z | - |
dc.date.available | 2018-05-29T04:51:18Z | - |
dc.date.issued | 2018-02 | - |
dc.identifier.other | 000000149662 | - |
dc.identifier.uri | https://hdl.handle.net/10371/142299 | - |
dc.description | 학위논문 (석사)-- 서울대학교 대학원 : 의과대학 의과학과, 2018. 2. 이승재. | - |
dc.description.abstract | Deposition of alpha-synuclein (αSyn) aggregates is a pathological feature of Parkinsons disease. Cell-to-cell transmission of αSyn aggregates was suggested as the underlying mechanism of the progression of Parkinsons disease. According to the prion-like spreading hypothesis, aggregate transmission increases dependently on the template seeding mechanism. αSyn aggregates work like infectious prion, amplifying protein aggregation and accumulation. However, this still remains unclear. V40G variant of αSyn has seeding blocking property | - |
dc.description.abstract | thereby elucidation of template seeding hypothesis becomes possible. Here, αSyn aggregate transmission rate was observed using Bimolecular Fluorescence Complementation (BiFC) system to verify whether this seeding ability plays a major role in αSyn aggregate propagation. Aggregate transmission level was increased in the V40G aggregate treated condition even the V40G aggregate has no seeding activity. Therefore, αSyn aggregate transmission might involve the mechanism other than the direct template seeding. To elucidate the mechanism of transmission, I assessed the endo-lysosomal degradation rate using fluorescein-conjugated dextran. When the cells were exposed to the extracellular αSyn, the endo-lysosomal degradation rate was decreased, while the lysosomal integrity and function were unaffected. RNA sequencing analysis revealed that the pathways related to the transcriptional regulation, cytoskeleton organization, immune response, and mitochondria were changed by extracellular αSyn aggregates. Altogether, this study suggests that the templated seeding mechanism is not the main principle of αSyn aggregate propagation and that the mechanism is related with the changes in the trafficking through the endo-lysosomal pathway. | - |
dc.description.tableofcontents | INTRODUCTION 1
Clinical and pathological features of Parkinsons disease 1 Cell-to-cell transmission of αSyn aggregates in Parkinsons disease 1 Structure of αSyn 4 Characteristics of V40G variant of αSyn 4 The purpose of the study 6 MATERIALS & METHODS 9 1. Materials 9 2. Cell culture and Stable cell line 10 3. Purification of recombinant wild type and V40G mutant αSyn and Fibril preparation 10 4. Thioflavin T binding assay 11 5. Circular dichroism spectroscopy 11 6. Seeding assay in Venus cell-lines 12 7. Immunofluorescent Cell staining 12 8. Galectin-3 transfection using electroporation 12 9. Characterization of lysosomal dysfunction 13 10. Preparation of cell extracts and lysosome fraction 13 11. Immunoblotting 14 12. Quant RNA-seq analysis 14 13. Network analysis 16 14. Quantification and Statistical analysis 17 RESULTS 18 The template seeding may not be the crucial mechanism of the intercellular aggregate propagation 18 Aged αSyn did not affect the selective autophagy, nor the ER-Golgi biosynthetic pathway 18 Treatment with the aged αSyn reduced the rate of the endo-lysosomal degradation 23 Lysosomal integrity was intact after the incubation with the aged αSyn 23 Lysosome acidification was intact 25 Reduction in the endo-lysosomal degradation was a long-term effect of the aged αSyn 28 Transcriptome analysis of the cells exposed to the aged aSyn 32 DISCUSSION 46 REFERENCES 50 국문 초록 57 | - |
dc.format | application/pdf | - |
dc.format.extent | 2323032 bytes | - |
dc.format.medium | application/pdf | - |
dc.language.iso | en | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject | Parkinson’s disease | - |
dc.subject | Cell-to-cell transmission | - |
dc.subject | Protein aggregates | - |
dc.subject | Alpha-synuclein | - |
dc.subject | Endo-lysosomal pathway | - |
dc.subject | Transcriptomics | - |
dc.subject | Gene expression change | - |
dc.subject.ddc | 610.72 | - |
dc.title | Mechanism of Extracellular Aggregate-induced Alpha-synuclein Transmission | - |
dc.type | Thesis | - |
dc.description.degree | Master | - |
dc.contributor.affiliation | 의과대학 의과학과 | - |
dc.date.awarded | 2018-02 | - |
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