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Localization of Goα in the Developing and Adult Mouse Brain
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 서해영 | - |
| dc.contributor.author | Cha, Hye Lim | - |
| dc.date.accessioned | 2019-10-21T07:19:51Z | - |
| dc.date.available | 2019-10-21T07:19:51Z | - |
| dc.date.issued | 2013-02 | - |
| dc.identifier.other | 13539 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/18200 | - |
| dc.description | 학위논문(석사)아주대학교 일반대학원 :의생명학과,2013. 2 | - |
| dc.description.tableofcontents | TABLE OF CONTENS ABSTRACTS ⅰ TABLE OF CONTENTS ⅱ LIST OF FIGURES ⅳ LIST OF TABLE ⅴ Ⅰ. INTRODUCTION 1 1. Heterotrimeric Guanine nucleotide-binding protein alpha subunit (Gnαo) 1 2. Localization of Goα subunit in the brain 2 3. Cerebellar cortex 2 Ⅱ. MATERIALS AND METHODS 6 1. Animals 6 2. Tissue preparation (for immunohistochemistry) 6 3. Immunohistochemistry 6 4. Cell cultures 7 4.1. Cerebellar neuron/Purkinje cell culture 7 4.2. Hippocampal neuron culture 7 5. EGFP-IRES3-Goα-CL, GFP-Akt-PH, GFP-PLCδ-PH Construct 8 6. Transfection 8 6.1. 293T cell transfection 8 6.2. F11/COS cell transfection and Forskolin treatment 8 6.3. Hippocampal neuron transfection 9 7. Western blot 9 8. Immunocytochemistry 10 9. In situ hybridization 11 9.1 In vitro RNA transcription 11 9.2 Tissue preparation (for in situ hybridization) 11 9.3 In situ hybridization 12 Ⅲ. RESULTS 15 Ⅳ. DISCUSSION 29 Ⅴ. CONCLUSION 33 REFERENCES 34 국문 요약 38 |LIST OF FIGURES Figure. 1. Structure of the cerebellar cortex 5 Figure. 2. Construct map of EGFP-IRES3- Goα-CL 13 Figure. 3. In situ hybridization probe for Goα 14 Figure. 4. Goα is abundant in the molecular and granule cell layers. 19 Figure. 5. Expression of Goα is neuron specific 20 Figure. 6. Neuron-specific expression of Goα in in vitro cultured cerebellar neurons. 21 Figure. 7-1. Subcellular distribution of Goα in Purkinje cells 22 Figure. 7-2. Subcellular compartmentalization of Goα and calbindin in Purkinje dendrites. 23 Figure. 8. Expression of Goα mRNA in Purkinje cells 24 Figure. 9. Goα does not affect the cytogenesis of cerebellar cortex. 25 Figure. 10. Expression of Goα in transfected 293T is plasma membrane targeting. 26 Figure. 11. Expression of Goα is plasma membrane dominant 27 Figure. 12. Distribution of Goα merges with GFP-Akt-PH, GFP-PLCδ-PH 28 |LIST OF TABLE Table.1. Comparison of Goα distribution in the cerebellum 4 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | Localization of Goα in the Developing and Adult Mouse Brain | - |
| dc.title.alternative | 차혜림 | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | 차혜림 | - |
| dc.contributor.department | 일반대학원 의생명과학과 | - |
| dc.date.awarded | 2013. 2 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 570860 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000013539 | - |
| dc.subject.keyword | Go protein | - |
| dc.subject.keyword | cerebellum | - |
| dc.subject.keyword | Purkinje cell | - |
| dc.subject.keyword | plasma membrane | - |
| dc.description.alternativeAbstract | -Abstracts- Localization of Goα in the Developing and Adult Mouse Brain Heterotrimeric G proteins mediate signal transduction generated by hormone and neurotransmitters. Among G proteins, Go, a member of the Gi/o family, is the most abundant heterotrimeric G protein in the brain but the roles and effectors for the Go have not been clearly defined. As previously reported, the expression of Go protein is found in various brain regions, including cerebrum, hippocampus, thalamus, and cerebellum. Although Go is already known to be present on both plasma- and endo-membranes, the precise subcellular location of Go protein has been a controversial issue because diverse detection methods such as in situ hybridization versus and immunohistochemistry yielded apparently contradictory results. We previously found that Go functions as a signaling scaffolding molecule that determines the subcellular localization of cAMP-dependent protein kinase A (Ghil et al., 2006). In this study, we showed that Go is expressed in Purkinje cell dendrites of developing and adult cerebellum by comparing the Go expression in wild type and Go knockout mice and confirm the subcellular location of Go protein through the hippocampal neuron in vitro culture. Consequently, this study provides more accurate information to speculate the role of Go protein on the basis of its correct localization as a character of the Go. | - |
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