1997 Fiscal Year Annual Research Report

シナプス伝達受容分子と可塑性

Research Project

Project/Area Number	07278102
Research Institution	The University of Tokyo
Principal Investigator	三品昌美東京大学, 大学院・医学系研究科, 教授 (80144351)
Co-Investigator(Kenkyū-buntansha)	狩野方伸理化学研究所, 脳科学総合センター, チームリーダー(研究 (40185963) 真鍋俊也東京大学, 大学院・医学系研究科, 講師 (70251212) 渡辺雅彦北海道大学, 医学部, 助教授 (70210945) 梅森久視東京大学, 医科学研究所, 助手 (20242117) 八木健岡崎国立共同研究機構, 生理学研究所研, 助教授 (10241241)
Keywords	LTP / グルタミン酸受容体 / NMDA受容体チャネル / リン酸化 / 小脳プルキンエ細胞 / シナプス形成 / 神経回路網 / G蛋白質
Research Abstract	本重点領域研究では、専門分野を異にする班員が協力して統合的な共同研究体制を組むことに成功し、一体となって以下に述べる成果を挙げてきた。1)海馬を含む前脳からシナプス後肥厚(PSD)分画を単離し、カルシウムとカルモデュリンを投与して内在性のCaMKIIを活性化することにより、PSDい共存するAMPA受容体チャネルがリン酸化されることを明らかにした。この結果から、シナプス長期増強(LTP)にCaMKIIによるAMPA受容体チャネルのリン酸化が関与する可能性が示唆された。2)NMDA受容体チャネルにカルモデュリンが結合するとチャネルは不活化される。この結合は、NMDA受容体チャネルがPKCによってリン酸化されると阻害された。またNMDA受容体チャネルはチロシンリン酸化により活性化されるが、このリン酸化は、PSD95を介してFynチロシンキナーゼが行っていた。このように、蛋白質相互作用とリン酸化によってNMDA受容体チャネル活性が制御されていることを明らかにした。3)海馬CA3野錐体細胞の頂上樹状突起に存在する交連/連合繊維入力によるシナプスでは、NMDA受容体チャネルε1サブユニットの欠損によりNMDA受容体応答およびLTPが低下した。一方、基底樹状突起に存在するfimbrial(Fim)入力によるシナプスでは、ε2サブユニットを片方の染色体で欠失させたヘテロ接合マウスでNMDA受容体応答およびLTPが低下することを見い出した。すなわち、海馬CA3野錐体細胞は多様なNMDA受容体チャネルの分子種をシナプスにより使い分けていることが示唆された。4)小脳プルキンエ細胞に特異的に発現するグルタミン酸受容体チャネルδ2サブユニット欠損マウスでは、プルキンエ細胞の産生や細胞移動は正常に起こり、樹状突起の形態分化や棘突起の形成も正常に起きるが、δ2サブユニット欠損は棘突起と平行線維終末との間の結合率を選択的に低下させることを見い出した(結合率:野生型100%、ノックアウト63%)。すなわち、δ2サブユニットは発達段階における平行線維シナプスの結合強化・安定化に関与することが明らかとなった。5)代謝型グルタミン酸受容体によるG蛋白質の活性化は、G蛋白質のαサブユニットのチロシンリン酸化によって制御されていることを見い出した。6)3量体GタンパクのGqのαサブユニット(Gαq)のノックアウトマウスの登上線維発達を調べ、生後3週間以後に約1/3のプルキンエ細胞で登上線維の多重支配が残存することを発見した。Gαqが小脳登上線維シナプス成熟に必要であることが明らかになった。7)グリア細胞に特異的に発現するグルタミン酸トランスポーターGLT-1を欠損したマウスでは、癲癇発作や脳損傷の憎悪が見られ、また、海馬のシナプス間隙でのグルタミン酸濃度上昇が野生型よりも大きいことを明らかにした。8)セマフォリンIII欠損マウスは胎生12日において末梢神経の走行に異常を示した。すなわち、動眼神経以外の三叉神経、顔面神経、迷走神経の軸索走行が拡散し、肢芽の軟骨組織や眼球上皮への神経の異常な進入が認められた。これらの結果から、セマフォリンIIIが生体内で神経回路網の形成に関与していることが明らかとなった。

Research Products
(40 results)

All Other

All Publications (40 results)

[Publications] Uchino,S.: "Inducible expression of N-methy-D-aspartate(NMDA)receptor channels from cloned cDNAs in CHO cells." Mol.Brain Res.44. 1-11 (1997)
[Publications] Uchino,S.: "Analysis of the glycine binding domain of the NMDA receptor channel ζ1 subunit." Neuro Report. 8. 445-449 (1997)
[Publications] Ito,I.: "Synapse-selective impairment of NMDA receptor functions in mice lacking NMDA receptor ε1 or ε2 subunit." J.Physiol.(Lond).500. 401-408 (1997)
[Publications] Kai,N.: "Molecular cloning of Fyn-associated molecules in the mouse central nervous system." J.Neurosci.Res.48. 407-424 (1997)
[Publications] Minami,T.: "Absence of prostglandin E_2-induced hyperalgesia in NMDA receptor ε subunit knockout mice." Br.J.Pharmacol.120. 1522-1526 (1997)
[Publications] Hayashi,Y.: "Calcium and calmodulin-dependent phosphorylation of AMPA type glutamate receptor subunits by endogenous protein kinases in the postsynaptic density." Mol.Brain Res.46. 338-342 (1997)
[Publications] Kawamoto,S.: "Arginine-481 mutation abolishes ligand-binding of the AMPA-selective glutamate receptor channel α1 subunit." Mol.Brain Res.47. 339-344 (1997)
[Publications] Taniguchi,M.: "Disruption of semaphorin III/D gene causes severe abnormality in peripheral nerve projection." Neuron. 19. 519-530 (1997)
[Publications] Kurihara,H.: "Impaired parallel fiber-Purkinje cell synapse stabilization during cerebellar development of mutant mice lacking the glutamate receptor δ2 subunit." J.Neurosci.15. 9613-9623 (1997)
[Publications] Kitsukawa,T.: "Neuropilin Function as a Receptor for Semaphorin III/D,and Plays a Crucial Role in Peripheral Nerve Projection in Mice." Neuron. 19. 995-1005 (1997)
[Publications] Miyakawa,T.: "Fyn-kinase as a determinant of ethanol sensitivity:Relation to NMDA-receptor function." Sience. 278. 698-701 (1997)
[Publications] Asada,H.: "Cleft palate and decreased brain gamma-aminobutyric acid in mice lacking the 67 kDa isoform of glutamic acid decarboxylase." Proc.Natl.Acad.Sci.USA. 94. 6496-6499 (1997)
[Publications] Umemori,H.: "Activation of the G protein Gq/11 through tyrosine phosphorylation of the alpha subunit." Science. 276. 1878-1881 (1997)
[Publications] Hisatsune,C.: "Phosphorylation-dependent regulation of NMDA receptors by calmodulin." J.Biol.Chem.272. 20805-20810 (1997)
[Publications] Offermanns,S.: "Impaired motor coordination and persistent multiple climbing fiber innervati on of cerebellar Purkinje cells of mice lacking Gαq." Proc.Natl.Acad.Sci.USA. 94. 14089-14094 (1997)
[Publications] Shibata,T.: "Glutamate Transporter GLAST is Expressed in the Radial Glia-Astrocyte Lineage of Developing Mouse Spinal Cord." J.Neurosci.17. 9212-9219 (1997)
[Publications] Itoh,M.: "Expression of a glutamate transporter subtype,EAAT4,in the developing human cerebellum." Brain Res.767. 265-271 (1997)
[Publications] Maeno-Hikichi,Y.: "Structure and functional expression of the cloned mouse neuronalhigh-affinity glutamate transporter." Mol.Brain Res.48. 176-180 (1997)
[Publications] Tanaka,J.: "Extra-junctional localization of glutamate transporter EAAT4 at excitatory Purkinje cell synapses." Neuroreport. 8. 2461-2464 (1997)
[Publications] Kohyama-Koganeya,A.: "Molecular cloning of a diacylglycerol kinase isozyme predominantly expressed in rat retina." FEBS Lett.409. 258-264 (1997)
[Publications] Tanaka,K.: "Epilepsy and exacerbation of brain injury in mice lacking the glutamate transporter GLT-1." Science. 276. 1699-1702 (1997)
[Publications] Nakagawa,S.: "Prominent expression of nuclear hormone receptor RORα in Purkinje cells from early development." Neurosci.Res.28. 177-184 (1997)
[Publications] Takayama,C.: "Altered intracellular localization of glutamate receptor channel δ2 subunit in weaver and reeler Purkinje cells." Brain Res.745. 231-242 (1997)
[Publications] Yamada,K.: "Changes in expression and distribution of the glutamate transporter EAAT4 in developing mouse Purkinje cells." Neurosci.Res.27. 191-198 (1997)
[Publications] Kano,M.: "Persistent multiple climbing fiber innervation of cerebellar Purkinje cells in mice lacking mGluR1." Neuron. 18. 71-79 (1997)
[Publications] Minakami,R.: "Phosphorylation and calmodulin binding of the metabotropic glutamate receptor subtype 5(mGluR5)are antagonistic in vitro." J.Biol.Chem.272. 20291-20298 (1997)
[Publications] Ikeda,K.: "Functional coupling of the nociceptin/orphanin FQ receptor with the G-protein-activated K^+(GIRK)channel." Mol.Brain Res.45. 117-126 (1997)
[Publications] Taniguchi,M.: "Efficient production of Cre-mediated site-directed recombinants through the utilization of the puromycin resistance gene,pac.a transient gene-integration marker for ES cells." Nucleic Acids Res.26. 679-680 (1998)
[Publications] Kitazawa,H.: "Abnormal synaptic transmission in the olfactory bulb of Fyn-kinase deficient mice." J.Neurophysiol.79.1. 137-142 (1998)
[Publications] Hashimoto,K.: "Presynaptic origin of paired-pulse depression at climbing fibre to Purkinje cell synapses in the rat cerebellum." J.Physiol.(Lond.). 506. 391-405 (1998)
[Publications] Watanabe,M.: "Selective scarcity of NMDA receptor channel subunits in the stratum lucidum (mossy fiber-recipient layer)of the mouse hippocampal CA3 subfield." Eur.J.Neursci.(in press).
[Publications] Tabuchi,S.: "Lipid mediators modulate NMDA receptor currents in a Xenopus oocyte expression system." Neurosci.Lett.(in press).
[Publications] Ito,I.: "Distribution and development of NMDA receptor activities at hippocampal synapses examined using mice lacking the ε1 subunit gene." Neurosci.Res.(in press).
[Publications] Watanabe,M.: "Contrasting localization of NMDA receptor channel subunits between the stratum lucidum and other neuropil layers in the adult mouse hippocampus." Eur.J.Neurosci.(in press).
[Publications] Yoshida,Y.: "ANA,a novel member of Tob/BTG1 family,is expressed in the ventricular zone of the developing central nervous system." Oncogene. (in press).
[Publications] Watanabe,M.: "Distinct Expressions and Distributions of Phospholipase Cβ1-4 mRNAs in Developing and Adult Mouse Brains." Eur.J.Neurosci.(in press).
[Publications] Nakagawa,S.: "Cytological compartmentalization in the staggerer cerebellum,as revealed by calbindin immunohistochemistry for Purkinje cells." J.Comp.Neurol.(in press).
[Publications] Watase,K.: "Motor discoordination and increased susceptibility to cerebellar injury in GLAST mutant mice." Eur.J.Neurosci.(in press).
[Publications] Harada,T.: "Retinal glial cell glutamate transporter plays a crucial role in glutamate clearance." Proc.Natl.Acad.Sci.USA. (in press).
[Publications] Manabe,T.: "Two froms of hippocampal long-term depression,the counterpart of long-term potentiation." Rev.Neurosci.(in press).

1997 Fiscal Year Annual Research Report

シナプス伝達受容分子と可塑性

Principal Investigator

三品 昌美 東京大学, 大学院・医学系研究科, 教授 (80144351)

Research Products

[Publications] Uchino,S.: "Inducible expression of N-methy-D-aspartate(NMDA)receptor channels from cloned cDNAs in CHO cells." Mol.Brain Res.44. 1-11 (1997)

[Publications] Uchino,S.: "Analysis of the glycine binding domain of the NMDA receptor channel ζ1 subunit." Neuro Report. 8. 445-449 (1997)

[Publications] Ito,I.: "Synapse-selective impairment of NMDA receptor functions in mice lacking NMDA receptor ε1 or ε2 subunit." J.Physiol.(Lond).500. 401-408 (1997)

[Publications] Kai,N.: "Molecular cloning of Fyn-associated molecules in the mouse central nervous system." J.Neurosci.Res.48. 407-424 (1997)

[Publications] Minami,T.: "Absence of prostglandin E_2-induced hyperalgesia in NMDA receptor ε subunit knockout mice." Br.J.Pharmacol.120. 1522-1526 (1997)

[Publications] Hayashi,Y.: "Calcium and calmodulin-dependent phosphorylation of AMPA type glutamate receptor subunits by endogenous protein kinases in the postsynaptic density." Mol.Brain Res.46. 338-342 (1997)

[Publications] Kawamoto,S.: "Arginine-481 mutation abolishes ligand-binding of the AMPA-selective glutamate receptor channel α1 subunit." Mol.Brain Res.47. 339-344 (1997)

[Publications] Taniguchi,M.: "Disruption of semaphorin III/D gene causes severe abnormality in peripheral nerve projection." Neuron. 19. 519-530 (1997)

[Publications] Kurihara,H.: "Impaired parallel fiber-Purkinje cell synapse stabilization during cerebellar development of mutant mice lacking the glutamate receptor δ2 subunit." J.Neurosci.15. 9613-9623 (1997)

[Publications] Kitsukawa,T.: "Neuropilin Function as a Receptor for Semaphorin III/D,and Plays a Crucial Role in Peripheral Nerve Projection in Mice." Neuron. 19. 995-1005 (1997)

[Publications] Miyakawa,T.: "Fyn-kinase as a determinant of ethanol sensitivity:Relation to NMDA-receptor function." Sience. 278. 698-701 (1997)

[Publications] Asada,H.: "Cleft palate and decreased brain gamma-aminobutyric acid in mice lacking the 67 kDa isoform of glutamic acid decarboxylase." Proc.Natl.Acad.Sci.USA. 94. 6496-6499 (1997)

[Publications] Umemori,H.: "Activation of the G protein Gq/11 through tyrosine phosphorylation of the alpha subunit." Science. 276. 1878-1881 (1997)

[Publications] Hisatsune,C.: "Phosphorylation-dependent regulation of NMDA receptors by calmodulin." J.Biol.Chem.272. 20805-20810 (1997)

[Publications] Offermanns,S.: "Impaired motor coordination and persistent multiple climbing fiber innervati on of cerebellar Purkinje cells of mice lacking Gαq." Proc.Natl.Acad.Sci.USA. 94. 14089-14094 (1997)

[Publications] Shibata,T.: "Glutamate Transporter GLAST is Expressed in the Radial Glia-Astrocyte Lineage of Developing Mouse Spinal Cord." J.Neurosci.17. 9212-9219 (1997)

[Publications] Itoh,M.: "Expression of a glutamate transporter subtype,EAAT4,in the developing human cerebellum." Brain Res.767. 265-271 (1997)

[Publications] Maeno-Hikichi,Y.: "Structure and functional expression of the cloned mouse neuronalhigh-affinity glutamate transporter." Mol.Brain Res.48. 176-180 (1997)

[Publications] Tanaka,J.: "Extra-junctional localization of glutamate transporter EAAT4 at excitatory Purkinje cell synapses." Neuroreport. 8. 2461-2464 (1997)

[Publications] Kohyama-Koganeya,A.: "Molecular cloning of a diacylglycerol kinase isozyme predominantly expressed in rat retina." FEBS Lett.409. 258-264 (1997)

[Publications] Tanaka,K.: "Epilepsy and exacerbation of brain injury in mice lacking the glutamate transporter GLT-1." Science. 276. 1699-1702 (1997)

[Publications] Nakagawa,S.: "Prominent expression of nuclear hormone receptor RORα in Purkinje cells from early development." Neurosci.Res.28. 177-184 (1997)

[Publications] Takayama,C.: "Altered intracellular localization of glutamate receptor channel δ2 subunit in weaver and reeler Purkinje cells." Brain Res.745. 231-242 (1997)

[Publications] Yamada,K.: "Changes in expression and distribution of the glutamate transporter EAAT4 in developing mouse Purkinje cells." Neurosci.Res.27. 191-198 (1997)

[Publications] Kano,M.: "Persistent multiple climbing fiber innervation of cerebellar Purkinje cells in mice lacking mGluR1." Neuron. 18. 71-79 (1997)

[Publications] Minakami,R.: "Phosphorylation and calmodulin binding of the metabotropic glutamate receptor subtype 5(mGluR5)are antagonistic in vitro." J.Biol.Chem.272. 20291-20298 (1997)

[Publications] Ikeda,K.: "Functional coupling of the nociceptin/orphanin FQ receptor with the G-protein-activated K^+(GIRK)channel." Mol.Brain Res.45. 117-126 (1997)

[Publications] Taniguchi,M.: "Efficient production of Cre-mediated site-directed recombinants through the utilization of the puromycin resistance gene,pac.a transient gene-integration marker for ES cells." Nucleic Acids Res.26. 679-680 (1998)

[Publications] Kitazawa,H.: "Abnormal synaptic transmission in the olfactory bulb of Fyn-kinase deficient mice." J.Neurophysiol.79.1. 137-142 (1998)

[Publications] Hashimoto,K.: "Presynaptic origin of paired-pulse depression at climbing fibre to Purkinje cell synapses in the rat cerebellum." J.Physiol.(Lond.). 506. 391-405 (1998)

[Publications] Watanabe,M.: "Selective scarcity of NMDA receptor channel subunits in the stratum lucidum (mossy fiber-recipient layer)of the mouse hippocampal CA3 subfield." Eur.J.Neursci.(in press).

[Publications] Tabuchi,S.: "Lipid mediators modulate NMDA receptor currents in a Xenopus oocyte expression system." Neurosci.Lett.(in press).

[Publications] Ito,I.: "Distribution and development of NMDA receptor activities at hippocampal synapses examined using mice lacking the ε1 subunit gene." Neurosci.Res.(in press).

[Publications] Watanabe,M.: "Contrasting localization of NMDA receptor channel subunits between the stratum lucidum and other neuropil layers in the adult mouse hippocampus." Eur.J.Neurosci.(in press).

[Publications] Yoshida,Y.: "ANA,a novel member of Tob/BTG1 family,is expressed in the ventricular zone of the developing central nervous system." Oncogene. (in press).

[Publications] Watanabe,M.: "Distinct Expressions and Distributions of Phospholipase Cβ1-4 mRNAs in Developing and Adult Mouse Brains." Eur.J.Neurosci.(in press).

[Publications] Nakagawa,S.: "Cytological compartmentalization in the staggerer cerebellum,as revealed by calbindin immunohistochemistry for Purkinje cells." J.Comp.Neurol.(in press).

[Publications] Watase,K.: "Motor discoordination and increased susceptibility to cerebellar injury in GLAST mutant mice." Eur.J.Neurosci.(in press).

[Publications] Harada,T.: "Retinal glial cell glutamate transporter plays a crucial role in glutamate clearance." Proc.Natl.Acad.Sci.USA. (in press).

[Publications] Manabe,T.: "Two froms of hippocampal long-term depression,the counterpart of long-term potentiation." Rev.Neurosci.(in press).

三品昌美東京大学, 大学院・医学系研究科, 教授 (80144351)