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Roles of neuronal Apolipoprotein E in the propagation of α-synuclein
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 박상면 | - |
| dc.contributor.author | 강서준 | - |
| dc.date.accessioned | 2022-11-29T03:01:22Z | - |
| dc.date.available | 2022-11-29T03:01:22Z | - |
| dc.date.issued | 2022-08 | - |
| dc.identifier.other | 32108 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/21086 | - |
| dc.description | 학위논문(박사)--아주대학교 일반대학원 :의생명과학과,2022. 8 | - |
| dc.description.abstract | 파키슨병 주된 특징은 단백질 응집체라 불리는 루이소체 혹은 루이 신경돌기가 뇌에서 생성된다. 루이소체와 루이신경돌기를 구성하는 주요 구성 단백질은 알파 시뉴클레인이며 프라이온과 유사하게 세포간 전파가 일어난다. 세포간 전달이 파킨슨병 약화에 중요한 역할을 한다는 증거는 아직 밝혀지지 않았지만, 많은 연구자들이 세포간 전달기작연구를 진행하고있다. 본 연구에서는 신경세포성 아포지질단백질 E가 알파시뉴클레인의 세포간 전달을 조절하는지와 분자적기작연구를 시험관 모델 및 생채모델을 통해 확인하였다. 우리는 신경세포성 아포지질단백질E 결핍이 LRP-1과 LDLR 발현량 감소를 확인하였고, 알파시뉴클레인의 세포수용을 약화시켰으며, 샤페론성 오토파지 기능이 증가됨으로써 알파시뉴클레인 방출을 약화시키는 것을 확인하였다. 또한 우리는 섬유성 알파시뉴클레인을 주입한 아포지질단백질E를 제거한 쥐에 섬유성 알파시뉴클레인을 주입 후 확인 결과 세포간 전달이 감소되어있는 것을 관찰하였다. 이를 통해 본 연구는 알파시뉴클레인 세포간 전파의 분자적 메커니즘을 이해하는데 도움이 될 것이라 생각된다. | - |
| dc.description.tableofcontents | I. INTRODUCTION 1 A. Parkinson's disease 1 B. α-Synuclein 1 C. Prion-like propagation of α-synuclein in PD 2 D. Apolipoprotein E 3 E. Apolipoprotein E in PD 4 F. Low-density lipoprotein receptors 5 G. Chaperone-mediated autophagy 6 H. Aims 7 II. MATERIAL AND METHODS 8 A. Reagents and antibodies 8 B. Plasmids and generation of stable cell lines 9 C. Cell culture 10 D. Western blot 10 E. Confocal microscopy 11 F. Dual-chamber assay 11 G. Coculture assay 12 H. Reverse transcription-polymerase chain reaction (RT-PCR) 12 I. Cholesterol measurement 13 J. Preparation of conditioned media from SH-SY5Y cells 13 K. Measurement of α‐syn in culture media 14 L. α‐Syn Real-time Quaking-Induced Conversion (RT-QUIC) assay 14 M. Animals and intracerebral injection 14 N. Immunohistochemistry 15 O. Quantitative pSer129 α-syn pathology 16 P. Statistical analysis 16 III. RESULTS 18 PART A. Effects of ApoE on α-syn uptake 18 1. α-Syn induces ApoE expression in neurons 18 2. ApoE deficiency inhibits α-syn uptake in neurons 21 3. Overexpression of ApoE isoforms increases α-syn uptake in neurons 24 4. ApoE deficiency increases intracellular cholesterol levels 27 5. ApoE deficiency decreases LDLR and LRP-1 29 6. LDLR and LRP-1 affect the α-syn uptake 32 PART B. Effects of ApoE on α-syn propagation 36 1. ApoE deficiency inhibits α-syn propagation in neurons 36 2. Knockout of LDLR and LRP-1 inhibits α-syn propagation in neurons 41 PART C. Effects of ApoE on α-syn release 44 1. ApoE deficiency inhibits α-syn release in neurons 44 2. Overexpression of ApoE isoforms increases α-syn release in neurons 48 3. ApoE deficiency enhances chaperone-mediated autophagy but not macroautophagy in neurons 50 4. The propagation of α-syn is reduced in ApoE KO mice 54 IV. DISCUSSION 57 V. CONCLUSIONS 63 VI. REFERENCES 65 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | Roles of neuronal Apolipoprotein E in the propagation of α-synuclein | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Kang Seo-Jun | - |
| dc.contributor.department | 일반대학원 의생명과학과 | - |
| dc.date.awarded | 2022. 8 | - |
| dc.description.degree | Doctoral | - |
| dc.identifier.localId | 1254156 | - |
| dc.identifier.uci | I804:41038-000000032108 | - |
| dc.identifier.url | https://dcoll.ajou.ac.kr/dcollection/common/orgView/000000032108 | - |
| dc.subject.keyword | ApoE | - |
| dc.subject.keyword | Lewy body | - |
| dc.subject.keyword | Parkinson’s disease | - |
| dc.subject.keyword | α-synuclein propagation | - |
| dc.subject.keyword | α-synuclein release | - |
| dc.subject.keyword | α-synuclein uptake | - |
| dc.description.alternativeAbstract | The presence of protein inclusions, called Lewy bodies (LBs) and Lewy neurites (LNs), in the brain is the main feature of Parkinson’s disease (PD). Recent evidence that prion-like propagation of α-synuclein (α-syn), as a major component of LBs and LNs, plays an important role in the progression of PD has gained much attention, although the molecular mechanism remains unclear. In this study, we evaluated whether neuronal ApoE regulates the cell-to-cell transmission of α-syn and explored its molecular mechanism using in vitro and in vivo model systems. We demonstrated that neuronal ApoE deficiency attenuated both α-syn uptake and release by downregulating LRP-1 and LDLR expression and enhancing chaperone-mediated autophagy (CMA) activity, respectively, to contribute to α-syn propagation. In addition, we observed that α-syn propagation was attenuated in ApoE KO mice injected with pre-formed mouse α-syn fibrils. This study will help understand the molecular mechanisms of α-syn propagation. | - |
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