Title

Glutamine-rich interactors of mutant huntingtin modulate neuronal toxicity by promoting cytoplasmic huntingtin aggregation

Alternative Title

독성 헌팅틴 단백질과 상호작용하는 글루타민 풍부 단백질의 세포질 헌팅틴 응집체 형성 촉진 및 신경세포 독성 조절 이해

DGIST Authors

In Jun Cha ; Sung Bae Lee ; Daehee Hwang

Advisor

이성배

Co-Advisor(s)

Daehee Hwang

Issued Date

2021

Awarded Date

2021/02

Citation

In Jun Cha. (2021). Glutamine-rich interactors of mutant huntingtin modulate neuronal toxicity by promoting cytoplasmic huntingtin aggregation. doi: 10.22677/thesis.200000363192

Type

Thesis

Subject

Huntington’s disease, Huntingtin, Cytoplasmic aggregates, Neuronal toxicity, 헌팅턴씨 병, 헌팅틴 단백질, 세포질 응집체, 신경세포 독성

Table Of Contents

Ⅰ. INTRODUCTION 1
ⅠⅠ. MATERIALS AND METHODS 4
2-1. Fly stocks 4
2-2. Generation of transgenic fly lines 5
2-3. Microscope image acquisition 5
2-4. Immunostaining 6
2-5. Image processing and analyses 6
2-6. Rapid Iterative Negative Geotaxis (RING) assay 7
2-7. Separation of SDS–soluble and –insoluble fractions for Western blot analysis 7
2-8. Nuclear extraction 8
2-9. Western blot 8
2-10. Statistical analysis 9
2-11. Analysis of survival rate 9
2-12. Identification of Q-rich proteins 9
2-13. Calculation of co-localization score (CLS) 10
2-14. Calculation of cytoplasm to nucleus score (CNS) 10
2-15. Partial least square-discriminatory analysis (PLS-DA) 11
2-16. Functional enrichment analysis 12
2-17. Reconstruction of network model 13
ⅠⅠⅠ. RESULTS 14
3-1. Conversion of cytoplasmic mHtt to cleaved nuclear mHtt is critical for mHtt toxicity 14
3-2. Knockdown of strong mHtt interactors reduces cytoplasmic mHtt aggregates 25
3-3. Cytoplasmic localization and Q-richness are important for strong interaction with cytoplasmic mHtt proteins 46
3-4. In silico model predicts cytoskeletal regulators as strong interactors to modulate the amounts of cytoplasmic mHtt aggregates 56
3-5. Knockdown of strong mHtt interactors increases mHtt toxicity in neurons 62
ⅠV. DISCUSSION 74
V. REFERENCES 78
VI. SUMMARY IN KOREAN 88

URI

http://dgist.dcollection.net/common/orgView/200000363192
http://hdl.handle.net/20.500.11750/16676

DOI

10.22677/thesis.200000363192

Degree

Doctor

Department

Brain and Cognitive Sciences

Publisher

DGIST

Show Full Item Record