| Project/Area Number |
12680789
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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 | Okazaki National Research Institutes |
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Principal Investigator |
YANAGAWA Yuchio Okazaki National Research Institutes Research Professor, 生理学研究所, 助教授 (90202366)
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| Co-Investigator(Kenkyū-buntansha) |
OBATA Kunihiko Okazaki National Research Institutes Professor, 生理学研究所, 教授 (60013976)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | GABAergic neuron / glutamate decarboxylase / GAD67 / GAD65 / knock-in mouse / label / GFP / cell migration / グルタミン酸脱炭酸酵素遺伝子 / トランスジェニックマウス / 標織 / プロモーター / lacZ遺伝子 |
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| Research Abstract |
In order to elucidate the molecular mechanisms of the development of GABAergic neurons, we labeled the GABAergic neurons with a reporter protein. First, we used transgenic mice to examine the expression pattern conferred by a 9.2-kb promoter-proximal DMA fragment of the glutamate decarboxylase (GAD) 65 gene fused to the bacterial /acZ reporter gene. Transgenic mice showed high β-galactosidase activity specifically in brain and testis. They also showed characteristic patterns of the transgene expression in olfactory bulb, cerebellar cortex and spinal cord, a similar expression pattern to that of endogenous GAD65. However, no transgene expression was observed in the ventral thalamus or hypothalamus, in which high GAD65 gene expression levels have been observed. These results suggest that the 9.2-kb DMA fragment of the GAD65 gene is associated with its tissue-specific expression and its targeted expression in GABAergic neurons of specific brain regions, but that additional regulatory elements are necessary to obtain fully correct expression. Second, we generated the GAD67-GFP mutant mice using knock-in method. In the GAD67-GFP mice, green fluorescence was observed in the GABAergic neurons without fixation. We utilized GAD67-GFP mice to observe distribution and migration of GABAergic neurons in the forebrain of the embryonic mouse. GFPpositve GABAergic neurons were found in the ganglionic eminences and the septum from Ell.5 and onwards. GABAergic neurons were also observed in the marginal zone and intermediate zone of the cortex. Using time-lapse imaging we observed that a large population of telencephalic GABAergic neurons migrate from the basal forebrain towards the cortex. These data suggest that GAD67-GFP mice will be useful for elucidating the development of GABAergic neurons.
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