Localization of Goα in the Developing and Adult Mouse Brain : 발생 중인 신경세포와 성체 생쥐 뇌에서의 Goα 단백질 발현 위치

다국어 초록 (Multilingual Abstract)

-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.

목차 (Table of Contents)

• 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