| Project/Area Number |
11680795
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | tyrosine hydroxylase / catecholamine / dopaminergic neuron / Cre-ERT / Cre-loxP / 相同組み換え / 条件付き遺伝子破壊 |
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| Research Abstract |
In this project, we plan to knock-out tyrosine hydroxylase at the desired time at the specific organ using the inducible knockout system with Cre-ERT.We constructed a floxed gene in which a crucial exon of thetyrosine hydroxylase gene was flanked by two loxP sites. We are transfecting the gene into ES cells to obtain homologously recombinant clones. We succeeded to obtain homologous recombinants of ES cells, to make chireric mice with the clones, and to produce floxed mice, which have two loxP sequences surrounding exons of tyrosine hydroxylase gene.
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 tyrosine hydroxylase gene.
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