Investigation of signal transduction in cerebellar Purkinje cell by in vivo expression of pertusis toxin.
| Project/Area Number |
16300099
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neuroscience in general
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| Research Institution | Kobe University |
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Principal Investigator |
AIBA Atsu Kobe University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (20271116)
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| Co-Investigator(Kenkyū-buntansha) |
HARADA Takeshi Kobe University, Graduate School of Medicine, Research Associate, 大学院・医学系研究科, 助手 (30362768)
松田 育雄 神戸大学, 大学院・医学系研究科, 助手 (50335452)
新石 健二 神戸大学, 大学院・医学系研究科, 助手 (40362769)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2005: ¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 2004: ¥8,500,000 (Direct Cost: ¥8,500,000)
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| Keywords | Purkinje cell / Gi / o / pertusis toxin / L7 / transgenic mouse / insulator / RGS |
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| Research Abstract |
In order to investigate the role of trimetric GTP binding proteins in cerebellar Purkinje cell, we generated two different kind of transgenic mice.
First we generated transgenic mice which express pertusis toxin (PTX) in Purkinje cells to inhibit the signal via a Gi/o family. Secondly, we generated mutant mice which lack RGS8 that has a GTPase activating domein for Gi/o family and is expressed prominently in Purkinje cells. We generated L7-PTX transgenic mice by microinjection of PTX gene driven by Purkinje cell specific L7 promoter. One L7-PTX transgenic mouse expressed PTX mRNA in cerebellum and showed reduction in spontaneous movement. The transgenic mouse died at 7 weeks old. Other transgenic mice expressed little PTX mRNA in the cerebellum and transgene were not transmitted to the offspring. These results suggested that expression of PTX in the cerebellum results in reduced viability. To overcome this situation, we generated a transgenic vector in which PTX expression was suppressed by doxycycline. We generated RGS8 knockout mice (KO) by gene targeting. We introduced loxP sites and neomycin resistant gene flanked by two FRT sites. We generated RGS8 mutant carrying loxP sequence with intact exons as well as the RGS8 KO mice in which two exons were deleted. RGS8 KO showed normal motor coordination, spontaneous movement and body weight. However, body temperature of RGS8 KO was lower than that of wild-type littermate. Furthermore, hypothermia induced by GABAB receptor agonist baclofen was mimicked in RGS8 KO mice. These results suggested that RGS8 regulates body temperature by controlling GABAB receptor signaling.
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Report
(3 results)
Research Products
(15 results)
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[Journal Article] Farnesylation of retinal transducin underlies its translocation during light adaptation2005
Author(s)
Kassai H, Aiba A, Nakao K, Nakamura K, Katsuki M, Xiong WH, Yau KW, Imai H, Shichida Y, Satomi Y, Takao T, Okano T, Fukada Y.
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Journal Title
Neuron 47(4)
Pages: 529-539
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] A gene-targeted mouse model for chorea-acanthocytosis.2005
Author(s)
Tomemori Y, Ichiba M, Kusumoto A, Mizuno E, Sato D, Muroya S, Nakamura M, Kawaguchi H, Yoshida H, Ueno S, Nakao K, Nakamura K, Aiba_A, Katsuki M, Sano A.
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Journal Title
J.Neurochem. 92(4)
Pages: 759-766
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Book] 脳はどこまでわかったか2005
Author(s)
井原康夫
Total Pages
310
Publisher
朝日新聞社
Description
「研究成果報告書概要(和文)」より
Related Report
