2000 Fiscal Year Annual Research Report

発達脳におけるシナプス淘汰と機能成熟に関与する細胞間情報伝達物質

Research Project

Project/Area Number	12053226
Research Category	Grant-in-Aid for Scientific Research on Priority Areas (A)
Research Institution	Kanazawa University
Principal Investigator	狩野方伸金沢大学, 医学部, 教授 (40185963)
Co-Investigator(Kenkyū-buntansha)	田端俊英金沢大学, 医学部, 助手 (80303270)
Keywords	生後発達 / 小脳 / プルキンエ細胞 / 登上線維 / 多重支配 / シナプス除去 / 一酸化窒素 / NO合成酵素
Research Abstract	本研究では、発達期小脳でみられる登上線維シナプス除去と機能成熟をモデルとして、機能的神経回路形成に関与する新たな細胞間情報伝達物質を同定することを目標にしている。このため、ethylene-vinylacetate copolymer(Elvax)法を用いて、発達期マウス小脳に種々の薬物を持続投与する実験を行なった。薬物とElvaxを溶媒によって混合し、200-400μm程度のフィルム状に引き伸し、500x500μm程度の微小ブロックを作製した。これを、生後10日前後のマウスの小脳表面に植え込んだ後に、マウスを生後30日程度まで成長させ、小脳スライスを作製した。プルキンエ細胞からwhole-cell patch-clampを行ない、登上線維を電気刺激して、プルキンエ細胞が何本の登上線維によって支配されているか、またそのシナプス伝達の性質について調べた。本年度は、まず、一酸化窒素(NO)から、soluble guanylate cyclaseとcGMP-dependent protein kinase(G kinase)につながる系に注目し、NO合成酵素(NOS)の非選択的ブロッカーであるL-NAMEとsoluble guanylate cyclaseのブロッカーであるODQを持続投与し、その効果を調べた。しかし、これまでにL-NAMEやODQの持続投与が登上線維シナプス除去に影響するという事実は得られていない。神経型NOSのノックアウトマウスにおいても、登上線維シナプス除去に異常はみられなかった。また、一方、インシュリン様増殖因子(IGF-1)とその受容体を介して細胞内チロシンリン酸化に至る系に関しては、IGF-1の持続投与により、登上線維によって多重支配されるプルキンエ細胞の比率が増大するという予備的な結果が得られた。今後、更なる検討を加える予定である。

Research Products
(14 results)

All Other

All Publications (14 results)

[Publications] Ichise,T.: "mGluR1 in cerebellar Purkinje cells essential for long-term depression, synapse elimination and motor coordination."Science. 288. 1832-1835 (2000)
[Publications] Tsubokawa,H.: "Calcium-dependent persistent facilitation of spike backpropagation in the CA1 pyramidal neurons."J.Neurosci.. 20. 4878-4884 (2000)
[Publications] Kakizawa,S.: "Critical period for activity-dependent synapse elimination in developing cerebellum."J.Neurosci.. 20. 4954-4961 (2000)
[Publications] Furuya,S.: "L-Serine and glycine serve as major astroglia-derived trophic factors for cerebellar Purkinje neurons"Proc.Natl.Acad.Sci.USA.. 97. 11528-11533 (2000)
[Publications] Tabata,T.: "A reliable method for culture of dissociated mouse cerebellar cells enriched for Purkinje Neurons."J.Neurosci.Meth.. 104. 45-53 (2000)
[Publications] Miyata,M.: "Local calcium release in dendritic spines required for long-term synaptic depression."Neuron. 28. 233-244 (2000)
[Publications] Ohno-Shosaku,T.: "Heterosynaptic expression of depolarization-induced suppression of inhibition (DSI) in rat hippocampal cultures."Neurosci.Res.. 36. 67-71 (2000)
[Publications] Kobayashi,K.: "Modest Neuropsychological deficits caused by reduced noradrenaline metabolism in mice heterozygous for a mutated tyrosine hydroxylase gene."J.Neurosci.. 20. 2418-2426 (2000)
[Publications] Tanaka,J.: "Gq protein subunits Gq and G11 are localized at the postsynaptic extra-junctional membrane of cerebellar Purkinje cells and hippocampal pyramidal cells."Eur.J.Neurosci.. 12. 781-792 (2000)
[Publications] Matsuzawa,M.: "Formation of hippocampal synapses on patterned substrates of a laminin-derived synthetic peptide."Eur.J.Neurosci.. 12. 903-910 (2000)
[Publications] Hashimoto,K.: "Climbing fiber synapse elimination during postnatal cerebellar development requires signal transduction involving Gαq and phospholipase Cγ4."Prog.Brain Res.. 124. 31-48 (2000)
[Publications] Ohno-Shosaku,T.: "Endogenous cannabinoids mediate retrograde signals from depolarized postsynaptic neurons to presynaptic terminals."Neuron. 29(in press). (2001)
[Publications] Kano,M.: "Endogenous corticotropin-releasing factor (CRF) is required for the induction of cerebellar long-term depression."T.Kato, (ed) Elsevier/Tokyo. 4 (2000)
[Publications] 柿澤昌: "小脳脳神経科学イラストレイテッド、第2章 5"羊土社(森寿,真鍋俊也,渡辺雅彦,岡野栄之,宮川剛/編). 6 (2000)

2000 Fiscal Year Annual Research Report

発達脳におけるシナプス淘汰と機能成熟に関与する細胞間情報伝達物質

Principal Investigator

狩野 方伸 金沢大学, 医学部, 教授 (40185963)

Research Products

[Publications] Ichise,T.: "mGluR1 in cerebellar Purkinje cells essential for long-term depression, synapse elimination and motor coordination."Science. 288. 1832-1835 (2000)

[Publications] Tsubokawa,H.: "Calcium-dependent persistent facilitation of spike backpropagation in the CA1 pyramidal neurons."J.Neurosci.. 20. 4878-4884 (2000)

[Publications] Kakizawa,S.: "Critical period for activity-dependent synapse elimination in developing cerebellum."J.Neurosci.. 20. 4954-4961 (2000)

[Publications] Furuya,S.: "L-Serine and glycine serve as major astroglia-derived trophic factors for cerebellar Purkinje neurons"Proc.Natl.Acad.Sci.USA.. 97. 11528-11533 (2000)

[Publications] Tabata,T.: "A reliable method for culture of dissociated mouse cerebellar cells enriched for Purkinje Neurons."J.Neurosci.Meth.. 104. 45-53 (2000)

[Publications] Miyata,M.: "Local calcium release in dendritic spines required for long-term synaptic depression."Neuron. 28. 233-244 (2000)

[Publications] Ohno-Shosaku,T.: "Heterosynaptic expression of depolarization-induced suppression of inhibition (DSI) in rat hippocampal cultures."Neurosci.Res.. 36. 67-71 (2000)

[Publications] Kobayashi,K.: "Modest Neuropsychological deficits caused by reduced noradrenaline metabolism in mice heterozygous for a mutated tyrosine hydroxylase gene."J.Neurosci.. 20. 2418-2426 (2000)

[Publications] Tanaka,J.: "Gq protein subunits Gq and G11 are localized at the postsynaptic extra-junctional membrane of cerebellar Purkinje cells and hippocampal pyramidal cells."Eur.J.Neurosci.. 12. 781-792 (2000)

[Publications] Matsuzawa,M.: "Formation of hippocampal synapses on patterned substrates of a laminin-derived synthetic peptide."Eur.J.Neurosci.. 12. 903-910 (2000)

[Publications] Hashimoto,K.: "Climbing fiber synapse elimination during postnatal cerebellar development requires signal transduction involving Gαq and phospholipase Cγ4."Prog.Brain Res.. 124. 31-48 (2000)

[Publications] Ohno-Shosaku,T.: "Endogenous cannabinoids mediate retrograde signals from depolarized postsynaptic neurons to presynaptic terminals."Neuron. 29(in press). (2001)

[Publications] Kano,M.: "Endogenous corticotropin-releasing factor (CRF) is required for the induction of cerebellar long-term depression."T.Kato, (ed) Elsevier/Tokyo. 4 (2000)

[Publications] 柿澤昌: "小脳 脳神経科学イラストレイテッド、第2章 5"羊土社(森寿,真鍋俊也,渡辺雅彦,岡野栄之,宮川剛/編). 6 (2000)

狩野方伸金沢大学, 医学部, 教授 (40185963)

[Publications] 柿澤昌: "小脳脳神経科学イラストレイテッド、第2章 5"羊土社(森寿,真鍋俊也,渡辺雅彦,岡野栄之,宮川剛/編). 6 (2000)