Histochemical and Molecular Genetical Studies on the Development of Cholinergic Neurons
| Project/Area Number |
01570036
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
神経解剖学
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| Research Institution | Tokyo Metropolitan Institute for Neurosciences |
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Principal Investigator |
ICHIKAWA Tomoyuki (財)東京都神経科学総合研究所, 解剖発生学研究部門, 主事研究員 (90150193)
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| Co-Investigator(Kenkyū-buntansha) |
DEGUCHI Takeo (財)東京都神経科学総合研究所, 分子神経生物学研究部門, 参事研究員 (20073059)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1989: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Cholinergic neuron / Choline acetyltransferase / Development / Immunohistochemistry / In situ hybridization / Molecular genetics / 神経回路網形成 |
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| Research Abstract |
Choline acetyltransterase (ChAT), the biosynthetic enzyme for ACh, is currently the most specific marker for the cholinergic neuron. cDNAs for ChAT were isolated from spinal cords of rat, mouse and human.
To investigate the development of the cholinergic neuron histochemically, we introduced molecular biological techniques into our anatomical studies. One method involves expression of ChAT protein from cDNA for rat ChAT introduced into Echerichia coli and the production of antisera against the protein. Using an antiserum obtained, immunohistochemistry (IHC) was established. The second method involves the development of an in situ hybridization (ISH) histochemistry, which enables the precise localization and indentification of individual cells in which a particular gene is transcribed. We have succeeded in detecting mRNA for ChAT of rat and human.
Using IHC and ISH, we clarified the postnatal development of the cholinergic neuron in the rat CNS. During the course of this study, we found a transient appearance of ChAT-immunoreactive neurons in the layer VI of the neocortex between postnatal day 2 and day 13.
This phenomenon is suggested to be due to the change of phenotype in these neurons rather than the death or migration of the neurons.
We have also succeeded in detecting mRNA for ChAT in motoneurons in the spinal cord of the human embryo, and this study will be continued.
On the other hand, molecular genetical analyses of ChAT gene revealed that structures of mRNA differ between the fetal and adult mouse spinal cords and there are several forms of mRNAs generated by a differential splicing in the adult mouse spinal cord. In addition, we have succeeded in cloning cDNAfor human ChAT and demonstrated that it is far larger than those for rat and mouse ones.
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Report
(3 results)
Research Products
(12 results)
