| Project/Area Number |
11694322
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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 | Fujita Health University |
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Principal Investigator |
ICHINOSE Hiroshi Fujita Health University, Institute for Comprehensive Medical Science, Professor, 総合医科学研究所, 教授 (90192492)
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| Co-Investigator(Kenkyū-buntansha) |
OHYE Tamae Fujita Health University, Institute for Comprehensive Medical Science, Research Associate, 総合医科学研究所, 助手 (10247661)
INAGAKI Hidehito Fujita Health University, Institute for Comprehensive Medical Science, Research Associate, 総合医科学研究所, 助手 (70308849)
SUZUKI Takahiro Fujita Health University, Institute for Comprehensive Medical Science, Research Associate, 総合医科学研究所, 助手 (70298545)
ICHINOSE Chiho Fujita Health University, School of Medicine, Assistant Professor, 医学部, 講師 (10247653)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2000: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | tyrosine hydroxylase / nuclear receptor / Nurr1 / dopaminergic neuron / ドーパミン / 発達 / Cre-loxP / カテコールアミン |
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| Research Abstract |
Mice lacking Nurr1 failed to generate midbrain dopaminergic neurons, and die soon after birth. In order to dissect the molecular mechanism for Nurr1, we plan to knock-out Nurr1 and tyrosine hydroxylase, a rate-limiting enzyme in catecholamine biosynthesis, at the desired time at the specific organ using the inducible knockout system that is developed by Prof. Pierre Chambon and Dr. Daniel Metzger (our foreign collaborators).
We screened the mouse genomic library constructed in lambda phage vector or bacterial artificial chromosome (BAC). We have isolated a Nurr1 gene, and constructed the floxed gene in which a crucial exon of the Nurr1 gene was flanked by loxP sites. We have obtained homologously recombinant ES clones.
For the expression of the inducible recombinase Cre-ERT in dopaminergic neurons, a knock-in mouse line has been generated by introducing (via homologous recombination) Cre-ERT within the first exon of the tyrosine hydroxylase gene. Since tyrosine hydroxylase is a catecholamine-synthesizing enzyme, we expect that the mice specifically express Cre-ERT in tyrosine hydroxylase positive neurons. The mice were successfully created, and we are examining the expression of Cre-ERT by RT-PCR and Western blotting. We will cross the floxed mouse with mouse expressing Cre-ERT in catecholaminergic neurons, and examine the effect of inducible knock-out of the Nurr1 and tyrosine hydroxylase genes.
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