1995 Fiscal Year Annual Research Report

海馬苔状線維-CA3野シナプスにおける長期増強の分子機能の解明

Research Project

Project/Area Number	07278219
Research Institution	Kyoto University
Principal Investigator	下濱俊京都大学, 医学研究科, 助手 (60235687)
Co-Investigator(Kenkyū-buntansha)	前田武彦京都大学, 薬学部, 助手 (50271010) 佐藤公道京都大学, 薬学部, 教授 (80025709) 北村淳京都大学, 医学研究科, 教授 (10204976)
Keywords	長期増強現象 / 海馬 / パッチクランプ / グルタミン酸 / シナプス / AMPA / KA / パッチ電流 / CA3
Research Abstract	本研究の目的は、海馬苔状線維-CA3野錐体細胞シナプスにおける長期増強(LTP)発現機構を、シナプスレベルで明らかにすることである.これまで,海馬LTP発現機構についてはシナプス前か,あるいはシナプス後起源か議論されてきたが,一致した結果は得られていない.シナプス前起源であれば,LTP発現後,伝達物質遊離が増大しているから,これを検出することが直接的な証明となるであろう.そこで申請者らは海馬スライス上で、電気生理学的に伝達物質遊離を検出する系の確立を行った.若齢ラットより海馬スライスを作製し,直視下でCA3野錐体細胞からアウトサイド-アウトパッチクランプ(OC)を行った。このOC電極(パッチ電極)上のパッチ膜が、AMPA/KA型グルタミン酸受容体拮抗薬感受性のグルタミン酸応答性を有することを確認した。パッチ電極先端を苔状線維-CA3野錐体細胞シナプス領域であるCA3野透明層に置き、苔状線維の電気刺激により遊離されるグルタミン酸をパッチ膜上のAMPA/KA型受容体を介した電流応答(パッチ電流)として検出することに成功した。脳スライス上で、しかも単発の電気刺激で遊離される伝達物質をリアルタイムで検出したのは、国内、国外を通じてはじめてである。今後は、本手技による伝達物質遊離の検出と、ホールセルクランプ法によるシナプス後細胞(CA3錐体細胞)のグルタミン酸応答性の検出を同時に行う系を確立し、LTP発現機構のシナプス起源を解明する予定である。また、LTP発現に伴う形態上の可塑的変化、また遺伝子導入や翻訳遮断によるLTPに対する影響についての分子機構を解明するため、培養海馬スライスの実験系を確立する予定である。

Research Products
(14 results)

All Other

All Publications (14 results)

[Publications] Maeda T: "Patch sensor detection of glutamate relense evoked by a single electrical shock" Neuron. 15. 253-257 (1995)
[Publications] Shimohawa S: "Glutamate-induced antigenic changes of phospholipcae C-δ in cultured cortical neurons" Journal of Neuroscience Research. 41. 418-426 (1995)
[Publications] Shimohama S: "Alterations of low molecular weight acid phosphatase in Abheimer's disease" Brain Research. 699. 125-129 (1995)
[Publications] Shimohama S: "Ditterential involvement of phospholipase C isozymes in Alzhoiner's disease" Gerontology. 41(suppl 1). 13-19 (1995)
[Publications] Shimohama S: "Signal trnsduction mechanisms in Alzheimer's disease" Alzheimer Disease and Associated Disorders. 9(Suppl 2). 15-22 (1995)
[Publications] Matsushima H: "Changes in platelet phosphohipase C protein levels and activity in Alzheimer's disease" Neurobiology of Aging. 16. 895-900 (1995)
[Publications] Shimohama S: "Nicotine-induced protection against glutamate cytotoxicity:Nicotinic cholinergic receptor-mediated inhibition of nitric oxide formation." Annals of New York Academy of Sciences. 777. 356-361 (1996)
[Publications] Sawada H: "Methylp honyl pyridium ion (MPP^+) enhances glutamate-induced cytotoxicity against do paminergic neurons in cultured rat mesencephalon." Journal of Neuroscince Research. 43. 55-62 (1996)
[Publications] Shimohama S: "Chauges of neurocalcin,a calcium-binding protein,in the brain of patieuts with Alzheimer's disease" Brain Research. (in press). (1996)
[Publications] Sawada H: "pitterent mechanisms of glutamate-induced neursnal death between dopsinergic and non-clopaminergic heurons in rat messencephalic culture." Journal of Neuroscience Research. (in press). (1996)
[Publications] Kawamata J: "A novel Gly16 Ser (GGC-AGC) mutation in the SOD-1 gene in a patient with appareutly spradic gouing-onset amyotrophic lateral sclerosis." Human Mutation. (in press). (1996)
[Publications] Matsushima H: "Ca^<3+>-dependent and-independent protein kinase C chouges in the brain of patients with Alzheimer's disease" Journal of Nourochemistry. (in press). (1996)
[Publications] Kawamata J: "Anyotrsphic Lateral Sclerosis-Progress and Perspectives in Basic Research and Clinical Application-" Doletion and point mutations in superoxide dismutase-1 geue in amyotrophis lateral sclerosis, (1996)
[Publications] Imura T: "Amyotrophic Lateral Sclerosis-Progress and Perspectives in Basic Research and Clinical Application-" Polymorphism in NF-H and Gp×1 genes in Japanese sporadicand fomilial ALS, (1996)

1995 Fiscal Year Annual Research Report

海馬苔状線維-CA3野シナプスにおける長期増強の分子機能の解明

Principal Investigator

下濱 俊 京都大学, 医学研究科, 助手 (60235687)

Research Products

[Publications] Maeda T: "Patch sensor detection of glutamate relense evoked by a single electrical shock" Neuron. 15. 253-257 (1995)

[Publications] Shimohawa S: "Glutamate-induced antigenic changes of phospholipcae C-δ in cultured cortical neurons" Journal of Neuroscience Research. 41. 418-426 (1995)

[Publications] Shimohama S: "Alterations of low molecular weight acid phosphatase in Abheimer's disease" Brain Research. 699. 125-129 (1995)

[Publications] Shimohama S: "Ditterential involvement of phospholipase C isozymes in Alzhoiner's disease" Gerontology. 41(suppl 1). 13-19 (1995)

[Publications] Shimohama S: "Signal trnsduction mechanisms in Alzheimer's disease" Alzheimer Disease and Associated Disorders. 9(Suppl 2). 15-22 (1995)

[Publications] Matsushima H: "Changes in platelet phosphohipase C protein levels and activity in Alzheimer's disease" Neurobiology of Aging. 16. 895-900 (1995)

[Publications] Shimohama S: "Nicotine-induced protection against glutamate cytotoxicity:Nicotinic cholinergic receptor-mediated inhibition of nitric oxide formation." Annals of New York Academy of Sciences. 777. 356-361 (1996)

[Publications] Sawada H: "Methylp honyl pyridium ion (MPP^+) enhances glutamate-induced cytotoxicity against do paminergic neurons in cultured rat mesencephalon." Journal of Neuroscince Research. 43. 55-62 (1996)

[Publications] Shimohama S: "Chauges of neurocalcin,a calcium-binding protein,in the brain of patieuts with Alzheimer's disease" Brain Research. (in press). (1996)

[Publications] Sawada H: "pitterent mechanisms of glutamate-induced neursnal death between dopsinergic and non-clopaminergic heurons in rat messencephalic culture." Journal of Neuroscience Research. (in press). (1996)

[Publications] Kawamata J: "A novel Gly16 Ser (GGC-AGC) mutation in the SOD-1 gene in a patient with appareutly spradic gouing-onset amyotrophic lateral sclerosis." Human Mutation. (in press). (1996)

[Publications] Matsushima H: "Ca^<3+>-dependent and-independent protein kinase C chouges in the brain of patients with Alzheimer's disease" Journal of Nourochemistry. (in press). (1996)

[Publications] Kawamata J: "Anyotrsphic Lateral Sclerosis-Progress and Perspectives in Basic Research and Clinical Application-" Doletion and point mutations in superoxide dismutase-1 geue in amyotrophis lateral sclerosis, (1996)

[Publications] Imura T: "Amyotrophic Lateral Sclerosis-Progress and Perspectives in Basic Research and Clinical Application-" Polymorphism in NF-H and Gp×1 genes in Japanese sporadicand fomilial ALS, (1996)

下濱俊京都大学, 医学研究科, 助手 (60235687)