1999 Fiscal Year Annual Research Report

興奮性シナプスに発現するクラスタリング分子の神経薬理学的研究

Research Project

Project/Area Number	11470025
Research Category	Grant-in-Aid for Scientific Research (B)
Research Institution	Kumamoto University
Principal Investigator	福永浩司熊本大学, 医学部, 助教授 (90136721)
Co-Investigator(Kenkyū-buntansha)	笠原二郎熊本大学, 医学部, 助手 (10295131) 山本秀幸熊本大学, 医学部, 講師 (60191433) 宮本英七熊本大学, 医学部, 教授 (50109659)
Keywords	シナプス / クラスタリング分子 / NMDA受容体 / カリウムチャネル / CaMキナーゼII / PSD-95 / SAP102 / PDZドメイン
Research Abstract	中枢神経系においてNMDA受容体,電位依存性カリウムチャネルのクラスタリング分子としてPSD-95/SAP90,NE-dlg/SAP102,chapsyn-110/PSD-93が知られている。本研究ではこれらのクラスタリング分子と機能的複合体を形成する細胞接着因子(neuroligin),NMDA受容体,カリウムチャネル,シグナル伝達分子(CaMキナーゼII,一酸化窒素合成酵素)の培養海馬神経細胞における,空間的,時間的な発現様式を調べた。さらに,カルシウムなどのシグナル伝達系が,複合体形成に如何に関与するのか検討した。培養神経細胞において自発的シナプス活動が観察される2週間以降においてNE-dlgは樹状突起,スパインに顕著に発現してくる。樹状突起におけるNE-dlgの発現部位は点状に存在しており,NMDA受容体の発現部位と一致した。さらに,興奮性シナプス後肥厚部に発現するCaMキナーゼIIのアルファサブユニットの発現と一致することから,これらの機能分子はシナプス後肥厚部において複合体を形成することが示された。次に,ツーハイブリッドシステムにてNE-dlgの結合蛋白質を検索した。NE-dlgはカルモデュリンとカルシウム依存性に結合した。このカルモデュリンとの結合はNE-dlgのPDZドメインとNMDA受容体(NMDAR2B)のC末端との結合には影響しなかった。さらに,NE-dlgはPSD-95とヘテロ結合体を形成し,カルモデュリンはこの結合を強化した。NMDA受容体からのカルシウムシグナルはNMDA受容体のスパインでの集積に抑制的に働いている。カルモデュリンとNE-dlgとの相互作用はこのプロセスに関与していると考えられる。

Research Products

(17 results)

All Other

All Publications (17 results)

[Publications] M. Ohmitsu: "Phosphorylation of myristoylated alanine-rich protein kinase C substrate by mitogen-activated protein kinase in cultured rat hippocampal neurons following stimulation of glutamate receptors."J. Biol. Chem.. 274. 408-417 (1999)
[Publications] K. Matsumoto: "Cloning from insulinoma cells of synapsin I associated with insulin secretory granules."J. Biol. Chem.. 274. 2053-2059 (1999)
[Publications] N. Masuko: "Interaction of NE-dlg/SAP102, a neuronal and endocrine tissue-specific mambrane-associated guanylate kinase protein, with calmodulin and PSD-95/SAP90: A possible regulatory role in molecular clustering at synaptic sites."J. Biol. Chem.. 274. 5782-5790 (1999)
[Publications] J. Kasahara: "Differential effects of a calcineurin inhibitor on glutamate-induced phosphorylation of Ca^<2+>/calmodulin-dependent protein kinases in cultured rat hippocampal neurons."J. Biol. Chem.. 274. 9061-9067 (1999)
[Publications] H. Kuwahara: "A novel NE-dlg/SAP102-associated protein, p51-nedasin, related to the amidohydrolase superfamily, interferes with the association between NE-dlg/SAP102 and N-Methyl-D-aspartete receptor."J. Biol. Chem.. 274. 32204-32214 (1999)
[Publications] J. Liu: "Differential roles of Ca^<2+>/calmodulin-dependent protein kinase II and mitogenactivated protein kinase activation in hippocampal long-term protentiation."J. Neurosci. 19. 8292-8299 (1999)
[Publications] M. Morioka: "Activities of calcineurin and phosphatese 2A in the hippocampus after transient forebrain ischemia."Brain Res.. 828. 135-144 (1999)
[Publications] H. Kanasaki: "Mitogen-activated protein kinase activation by stimulation with thyrotropinreleasing hormone in rat pituitary GH3 cells."Biol. of Reprod.. 61. 319-325 (1999)
[Publications] I. Munir: "Mitogen-activated protein kinase activation and regulation of cyclooxygenese 2 expression by platelet-activating factor and hCG in human endometrial adenocarcinoma cell line HEC-1B."J. Reprod. Fertil.. 117. 49-59 (1999)
[Publications] Q. Zhang: "Changes of free fatty acids and acyl-coas in rat brain hippocampal slice with tetraethylammonium-induced long-term potentiation."Biochem. Biophys. Res. Commun.. 267. 208-212 (1999)
[Publications] K. Fukunaga: "Current studies on a working model of CaM kinase II in hippocampal long-term potentiation and memory."Jpn. J. Pharmacol.. 79. 7-15 (1999)
[Publications] 福永浩司: "海馬における神経活動依存性シナプス伝達効率の調節"蛋白質核酸酵素,増刊「神経回路形成と機能発達」. 45. 474-482 (2000)
[Publications] J. Kasahara: "Activation of Ca^<2+>/calmodulin-dependent protein kinase IV in cultured rat hippocampal neurons."J. Neurosci. Res.. 59. 594-600 (2000)
[Publications] K. Fukunaga: "Decreased protein phosphatase 2A activity in hippocampal long-term potentiation."J. Neurochem.. (in press). (2000)
[Publications] K. Fukunaga: "Inhibition of neuromal nitric oxide synthase activity by DY-9760e, a novel neuroprotective agent, in vitro and in cultured neuroblastoma cells in situ."Biochem. Pharmacol.. (in press). (2000)
[Publications] H. Kanasaki: "Involvement of p38 mitogen-activated protein kinase activation in bromocriptineinduced apoptosis in rat pituitary GH3 cells."Biol. of Reprod.. (in press). (2000)
[Publications] 宮本英七: "脳と神経:分子神経生物科学入門(編集:金子章道,川村光毅,植村慶一) 第3章神経情報の統合,5.脳可塑性"共立出版. 7 (1999)

1999 Fiscal Year Annual Research Report

興奮性シナプスに発現するクラスタリング分子の神経薬理学的研究

Principal Investigator

福永 浩司 熊本大学, 医学部, 助教授 (90136721)

Research Products

[Publications] M. Ohmitsu: "Phosphorylation of myristoylated alanine-rich protein kinase C substrate by mitogen-activated protein kinase in cultured rat hippocampal neurons following stimulation of glutamate receptors."J. Biol. Chem.. 274. 408-417 (1999)

[Publications] K. Matsumoto: "Cloning from insulinoma cells of synapsin I associated with insulin secretory granules."J. Biol. Chem.. 274. 2053-2059 (1999)

[Publications] N. Masuko: "Interaction of NE-dlg/SAP102, a neuronal and endocrine tissue-specific mambrane-associated guanylate kinase protein, with calmodulin and PSD-95/SAP90: A possible regulatory role in molecular clustering at synaptic sites."J. Biol. Chem.. 274. 5782-5790 (1999)

[Publications] J. Kasahara: "Differential effects of a calcineurin inhibitor on glutamate-induced phosphorylation of Ca^<2+>/calmodulin-dependent protein kinases in cultured rat hippocampal neurons."J. Biol. Chem.. 274. 9061-9067 (1999)

[Publications] H. Kuwahara: "A novel NE-dlg/SAP102-associated protein, p51-nedasin, related to the amidohydrolase superfamily, interferes with the association between NE-dlg/SAP102 and N-Methyl-D-aspartete receptor."J. Biol. Chem.. 274. 32204-32214 (1999)

[Publications] J. Liu: "Differential roles of Ca^<2+>/calmodulin-dependent protein kinase II and mitogenactivated protein kinase activation in hippocampal long-term protentiation."J. Neurosci. 19. 8292-8299 (1999)

[Publications] M. Morioka: "Activities of calcineurin and phosphatese 2A in the hippocampus after transient forebrain ischemia."Brain Res.. 828. 135-144 (1999)

[Publications] H. Kanasaki: "Mitogen-activated protein kinase activation by stimulation with thyrotropinreleasing hormone in rat pituitary GH3 cells."Biol. of Reprod.. 61. 319-325 (1999)

[Publications] I. Munir: "Mitogen-activated protein kinase activation and regulation of cyclooxygenese 2 expression by platelet-activating factor and hCG in human endometrial adenocarcinoma cell line HEC-1B."J. Reprod. Fertil.. 117. 49-59 (1999)

[Publications] Q. Zhang: "Changes of free fatty acids and acyl-coas in rat brain hippocampal slice with tetraethylammonium-induced long-term potentiation."Biochem. Biophys. Res. Commun.. 267. 208-212 (1999)

[Publications] K. Fukunaga: "Current studies on a working model of CaM kinase II in hippocampal long-term potentiation and memory."Jpn. J. Pharmacol.. 79. 7-15 (1999)

[Publications] 福永浩司: "海馬における神経活動依存性シナプス伝達効率の調節"蛋白質 核酸 酵素,増刊「神経回路形成と機能発達」. 45. 474-482 (2000)

[Publications] J. Kasahara: "Activation of Ca^<2+>/calmodulin-dependent protein kinase IV in cultured rat hippocampal neurons."J. Neurosci. Res.. 59. 594-600 (2000)

[Publications] K. Fukunaga: "Decreased protein phosphatase 2A activity in hippocampal long-term potentiation."J. Neurochem.. (in press). (2000)

[Publications] K. Fukunaga: "Inhibition of neuromal nitric oxide synthase activity by DY-9760e, a novel neuroprotective agent, in vitro and in cultured neuroblastoma cells in situ."Biochem. Pharmacol.. (in press). (2000)

[Publications] H. Kanasaki: "Involvement of p38 mitogen-activated protein kinase activation in bromocriptineinduced apoptosis in rat pituitary GH3 cells."Biol. of Reprod.. (in press). (2000)

[Publications] 宮本英七: "脳と神経:分子神経生物科学入門(編集:金子章道,川村光毅,植村慶一) 第3章 神経情報の統合,5.脳可塑性"共立出版. 7 (1999)

福永浩司熊本大学, 医学部, 助教授 (90136721)

[Publications] 福永浩司: "海馬における神経活動依存性シナプス伝達効率の調節"蛋白質核酸酵素,増刊「神経回路形成と機能発達」. 45. 474-482 (2000)

[Publications] 宮本英七: "脳と神経:分子神経生物科学入門(編集:金子章道,川村光毅,植村慶一) 第3章神経情報の統合,5.脳可塑性"共立出版. 7 (1999)