| Project/Area Number |
17300100
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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 | Gunma University (2006)
Kanazawa University (2005) |
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Principal Investigator |
HIRAI Hirokazu GUNMA UNIVERSITY, GRADUATE SCHOOL OF MEDICINE, PROFESSOR, 大学院・医学系研究科, 教授 (70291086)
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| Co-Investigator(Kenkyū-buntansha) |
YANAGIHARA Dai UNIVERSITY OF TOKYO, GRADUATE SCHOOL OF ARTS AND SCIENCES, 大学院・総合文化研究科, 助教授 (90252725)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥15,200,000 (Direct Cost: ¥15,200,000)
Fiscal Year 2006: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2005: ¥12,200,000 (Direct Cost: ¥12,200,000)
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| Keywords | Purkinje cell / Glutamate receptor / Transgenic mouse / GluR2 / Cerebellum |
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| Research Abstract |
Tropism of lentiviral vectors for Purkinje cells of a mouse in vivo is influenced by cultivating condition of HEK293T cells, host cells for viral production. The tropism for Purkinje cells is high when the pH of the culture medium is >7.2, whereas the lentiviral vectors produced in the medium with the pH of <7.0 has high tropism for Bergmann glia.
We aimed to produce gene-modified mice in which phosphorylation of GluR2 Ser880 is impaired specifically in Purkinje cells. This is attained by crossing 2 different transgenic mice; A mouse expressing L7-rtTA and that express GluR2 K882A under the control of tetracycline-responsive (TRE) promoter. Expression of rtTA in the former mouse is driven by Purkinje-cell-specific L7 promoter. The transgenic mouse having both transgenes expresses GluR2 K882A only when doxycycline is administered through water and food. We obtained 6 lines of mice with L7-rtTA transgene, in which only 2 lines showed rtTA expression in Purkinje cells. While 4 lines of mice with TRE promoter-GluR2 K882 and 3 lines of mice with TRE promoter-wild-type GluR2 were obtained. Now we are increasing the number of those mice for crossing them with L7-rtTA mice.
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