活動依存的カルシウム流入による競合的シナプス回路発達の共通原理の解明

2008 Fiscal Year Annual Research Report

• Project/Area Number: 19100005
• Research Institution: Hokkaido University
• Principal Investigator: 渡辺 雅彦 Hokkaido University, 大学院・医学研究科, 教授 (70210945)
• Co-Investigator(Kenkyū-buntansha): 崎村 建司 新潟大学, 脳研究所, 教授 (40162325)
• Keywords: シナプス / 脳 / 発達 / バレル / 大脳皮質 / カルシニューリン / 臨界期可塑性 / CaMキナーゼ

Research Abstract

カルシニューリンはカルシウム依存性の脱リン酸化酵素で、触媒サブユニットCNAと制御サブユニットCNBからなるヘテロダイマーとして機能する。脳ではCNAα, CNAβ, CNB1のみが発現しているため、CNB1の遺伝子欠失により脳におけるカルシニューリン活性をほぼ完全に消失させることができるが、そのnull型欠損CNB1マウスは致死的である。本年度は、大脳皮質特異的Creマウスとの交配により、終脳選択的CNB1ノックアウトマウスを作成した。現在、その解析を行っており、バレル形成・発達において形状やサイズが異なる表現型を見いだした段階である。来年度において、この表現型が一体どのようなシナプス回路の発達異常によるものかの確定を行う予定である。
昨年度、CaMKIIα, CaMKIIβ, CNB1に対するポリクローナル抗体作成に着手し、後二者に対する特異抗体の作成に成功した。これらの抗体を用いて、小脳と大脳における分子局在を共焦点レーザー顕微鏡による多重染色と包埋前/包埋後免疫電顕法を解析した。その結果、これらのカルシウム依存性リン酸化酵素および脱リン酸化酵素はニューロンやシナプスの各部に広く分布し、特にシナプス可塑性が起こりやすい大脳錐体細胞の興奮性シナプスや、小脳の平行線維・プルキンエ細胞シナプスに豊富である事が判明した。さらに、CNB1の免疫反応が特異的であることは、上記終脳選択的CNB1ノックアウトマウスを用いる事で証明できた。今後、これらの分子の小脳シナプス回路形成発達における機能的役割を、遺伝子欠損マウスを用いて解析を行う予定である。

Research Products

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• [Journal Article] IQ-ArfGEF / BRAG1 is aguanine nucleotide exchange factor for Arf6 that inte racts and localizes with PSD-95 at excitatory synapses (2008)
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• [Journal Article] SK2 channel plasticity contributes to LTP at Shaffer collateral-CA1 synapses (2008)
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• [Journal Article] Effects of FAK ablation on cerebellar foliation, Bergmann glia and climbing fiber territory on Purkinje cells (2008)
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• [Journal Article] Expression of AMPA receptor subunits at synapses in laminae I-III of the rodent spinal dorsal horn (2008)
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• [Journal Article] Cerebellar pathology in transgenic mice expressing the pseudorabies virus immediate-early protein IE180 (2008)
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• [Journal Article] Role of the internal Shank-binding segment of glutamate receptor δ2 in synaptic localization and cerebellar functions (2008)
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• [Journal Article] Accumulation of AMPA receptors in autophagosomes in neuronal axons lacking adaptor protein AP4 (2008)
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• [Journal Article] Functional contributions of synaptically localized NR2B subunits of the NMD A receptor to synaptic transmission and long-term potentiation in the adultmouse CNS (2008)
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• [Journal Article] Cell type-specific subunit composition of G-protein-gated potassium channeisin the cerebellum (2008)
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• [Journal Article] Glutamate transporters regulate lesion-induced period plasticity in the developing somatosensory cortex (2008)
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• [Journal Article] Dual coupling of T-type Ca2+ channels shapes sleep-related oscillations in thalamic dendrites (2008)
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• [Journal Article] Synapse formation and clustering of neuroligin-2 in the absence of GABAA receptors (2008)
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• [Journal Article] Cloning and characterization of E-dlg, a novel splice variant of mouse homologue of the Drosophila discs large tumor suppressor binds preferentially to SAP 102 (2008)
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• [Journal Article] Cbln1 Regulates Rapid Formation and Maintenance of Excitatory Synapses in Mature Cerebellar Purkinje Cells in vitro and in vivo (2008)
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• [Journal Article] N-Ethylmaleimide-sensitive fusion protein (NSF) is involved in central sensitization in the spinal cord through GluR2 subunit composition switch after inflammation (2008)
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• [Journal Article] Enhanced cerebellar LTD induction and OKR adaptation in mice lacking del philin (2008)
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• [Journal Article] Spinocerebellar ataxia type 6 knock-in mice develop a progressive neuronaldysfunction with age-dependent accumulation of mutant Cav2.1 channels (2008)
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• [Journal Article] Use-dependent amplification of presynaptic Ca^<2+> signaling by axonal ryanodine receptors at the hippocampal mossy fiber synapse (2008)
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• [Journal Article] Type 1 cannabinoid receptor (CB1)-containing axons innervate the hypophysi otropic thyrotropin-releasing hormone-synthesizing neurons (2008)
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• [Journal Article] Peripheral, but not central, CB1 antagonism provides food intake-independent metabolic benefits in diet-induced obese rats (2008)
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• [Journal Article] Expression and possible role of creatine transporter in the brain and at the blood-cerebrospinal fluid barrier as a transporting Protein of guanidinoacetate, an endogenous convulsant (2008)
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• [Journal Article] Loss of Hrs in the central nervous system causes accumulation of ubiquitinated proteins and neurodegeneration (2008)
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• [Journal Article] The endocannabinoid system is modulated in response to spinal cord injury in rats (2008)
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• [Journal Article] Predominant expression of phospholipase Cβ1 in telencephalic principal neur ons and cerebellar interneurons, and its close allocation with related signaling molecules in somatodendritic neuronal elements (2008)
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• [Journal Article] Phospholipase Cβ3 in mouse and human dorsal root ganglia and spinal cord is a possible target for treatment of neuropathic pain (2008)
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• [Journal Article] CB1 cannabinoid receptor-dependent and-independent inhibition of depolarization-induced calcium influx in oligodendrocytes (2008)
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• [Journal Article] Left-right asymmetry of the hippocampal synapses with different subunit allocation of qlutamate receptors (2008)
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• [Journal Article] Large projection neurons in laminal of the rat spinal cord that lack the neurokinin 1 receptor are densely innervated by VGLUT2-containing axons and possess GluR4-containing AMPA receptors (2008)
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• [Journal Article] Diacylglycerol kinase β accumulates on the perisynaptic site of medium spiny neurons in the Striatum (2008)
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• [Presentation] マウス小脳および海馬におけるセクレトグラニンII(SgII)の局在様式 (2009)
  • Author(s): 宮崎太輔・渡辺雅彦
  • Organizer: 第114回解剖学会総会・全国学術集会
  • Place of Presentation: 岡山理科大学
  • Year and Date: 20090328-30
• [Presentation] M1受容体を介するコリン作動性神経の伝達様式とシナプス伝達調節機構 (2009)
  • Author(s): 山崎美和子, 渡辺雅彦
  • Organizer: 第114回解剖学会総会・全国学術集会
  • Place of Presentation: 岡山理科大学
  • Year and Date: 20090328-30
• [Presentation] マウス歯状回における2-アラキドノイルグリセロールを介した逆行性伝達抑制機構に関与する分子の免疫組織化学的分布 (2009)
  • Author(s): 内ヶ島基政・渡辺雅彦
  • Organizer: 第114回解剖学会総会・全国学術集会
  • Place of Presentation: 岡山理科大学
  • Year and Date: 20090328-30
• [Presentation] マウス小脳ピンスーにおける化学的シナプス伝達機構の検討 (2009)
  • Author(s): 岩倉淳, 渡辺雅彦
  • Organizer: 第114回解剖学会総会・全国学術集会
  • Place of Presentation: 岡山理科大学
  • Year and Date: 20090328-30
• [Presentation] GLASTはバーグマングリアの形態分化および登上線維単一支配の維持に必要である (2008)
  • Author(s): 宮崎太輔, 渡辺雅彦
  • Organizer: 日本解剖学会第54回東北・北海道連合支部学術集会
  • Place of Presentation: 市民プラザ・ビッグアイ(郡山市)
  • Year and Date: 20080906-07
• [Presentation] M1アセチルコリン受容体はマウスの大脳皮質・海馬の投射ニューロンに豊富に発現している (2008)
  • Author(s): 山崎美和子, 渡辺雅彦
  • Organizer: 日本解剖学会第54回東北・北海道連合支部学術集会
  • Place of Presentation: 市民プラザ・ビッグアイ(郡山市)
  • Year and Date: 20080906-07
• [Presentation] GLAST is essential for the maintenance of climbing fiber monoinnervation through preventing iocal ectopic innervations to neighbouring and remote Purkinje cells and promoting cytological differentiation of Bergmann glia (2008)
  • Author(s): Miyazaki T, Yamasaki M, Kouichi Tanaka K, Watanabe M.
  • Organizer: 6^<th> FENS Forum of Europian Neuroscience
  • Place of Presentation: The Ceneva Palexpo Conference Center, Geneva. Switzerland
  • Year and Date: 20080712-16
• [Presentation] Preferential localization of muscarinic M1 receptor in spines and thin dendrites of cortical principal neurons and its anatomical evidence for volume transmission (2008)
  • Author(s): Yamasaki M, Watanabe M.
  • Organizer: 6^<th> FENS Forum of Europian Neuroscience
  • Place of Presentation: The Ceneva Palexpo Conference Center, Geneva, Switzerland
  • Year and Date: 20080712-16
• [Presentation] Distinct cellular and subcellular expression of calcineurins in the mouse cere bellum (2008)
  • Author(s): Miura E, Watanabe M.
  • Organizer: 6^<th> FENS Forum of Europian Neuroscience
  • Place of Presentation: The Ceneva Palexpo Conference Center, Geneva, Switzerland
  • Year and Date: 20080712-16
• [Presentation] Monoacylglycerol lipase is expressed not only in presynaptic terminals, but also in astrocvtes in the mouse forebrain (2008)
  • Author(s): Uchigashima M, Watanabe M.
  • Organizer: 6^<th> FENS Forum of Europian Neuroscience
  • Place of Presentation: The Ceneva Palexpo Conference Cente, Geneva, Switzerland
  • Year and Date: 20080712-16
• [Presentation] Glutamate transporters magnify critical period plasticity in developing somatosensory cortex (2008)
  • Author(s): Takasaki C, Okada R, Mitani A, Fukaya M, Hujihara Y, Shirakawa T, Kouichi Tanaka K, Watanabe M.
  • Organizer: 6^<th> FENS Forum of Europian Neuroscience
  • Place of Presentation: The Ceneva Palexpo Conference Center, Geneva, Switzerland
  • Year and Date: 20080712-16
• [Presentation] マウス小脳におけるプルキンエ細胞のPKCγバンド状発現 (2008)
  • Author(s): 江本美穂, 渡辺雅彦
  • Organizer: Neuro 2008 第31回日本神経科学大会
  • Place of Presentation: 東京国際フォーラム
  • Year and Date: 20080709-11
• [Presentation] 成体マウス大脳皮質におけるニューロン周囲オリゴデンドロサイトの化学的特性の検討 (2008)
  • Author(s): 高崎千尋・内ヶ島基政・渡辺雅彦
  • Organizer: 第114回解剖学会総会・全国学術集会
  • Place of Presentation: 岡山理科大学
  • Year and Date: 20080328-30
• [Presentation] マウス小脳ピンスーにおける化学的シナプス伝達機構の形態学的検討 (2008)
  • Author(s): 岩倉淳, 渡辺雅彦
  • Organizer: 2008年日本顕微鏡学会北海道支部会
  • Place of Presentation: 北海道大学
  • Year and Date: 2008-12-01