DGIST Scholar: Identification of intrinsic resilience factors in MNs to ALS-toxicity

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Title: Identification of intrinsic resilience factors in MNs to ALS-toxicity

Description: Amyotrophic lateral sclerosis, TDP-43 proteinopathy, chicken embryo, spinal cord, RNA-sequencing

Table Of Contents: List of Contents
Abstract i
List of contents ii
List of tables iv
List of figures v

Ⅰ. Introduction
1.1. Overview of ALS 1
1.2. TDP-43 pathology in ALS 1
1.3. TDP-43 Q331K 3
1.4. Current therapeutic limitations in ALS 4
1.5. Selective vulnerability and resilience of MNs in ALS 4
1.6. Chick embryo model 5
1.7. Study rationale and objective 5

II. Materials and Methods
2.1. Egg preparation 7
2.2. In ovo electroporation of chick embryos 7
2.3. Cell isolation 8
2.4. cDNA synthesis and pre-amplification 8
2.5. Quantitative real-time PCR 9
2.6. Gene synthesis and cloning 10
2.7. RNA-sequencing 11
2.8. Tissue collection 11
2.9. Immunohistochemistry 11
2.10. RNA in situ hybridization 12

III. Results
3.1. Establishment of a chick embryonic spinal cord model for TDP-43 Q331K-induced toxicity 14
3.2. Transcriptome analysis identifies candidate genes in Hb9-GFP-positive MNs under TDP-43 Q331K-induced toxicity 17
3.3. RNA in situ hybridization analysis of candidate genes in the chick embryonic spinal cord 20
3.4. Preliminary analysis of time-dependent trends in TDP-43 Q331K subcellular localization 22
3.5. Analysis of the effect of LOC430766 co-expression on TDP-43-positive cell number and subcellular localization 25
3.6. Analysis of the effect of ULK1 co-expression on TDP-43-positive cell number and subcellular localization 29
3.7. Optimization of an mRNA-based TDP-43 Q331K expression system 33

IV. Discussion 37
V. References 47
요약문 54

URI: https://scholar.dgist.ac.kr/handle/20.500.11750/60764
http://dgist.dcollection.net/common/orgView/200001007748

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