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

2007 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): 狩野 方伸 東京大学, 大学院・医学系研究科, 教授 (40185963) ; 崎村 建司 新潟大学, 脳研究所, 教授 (40162325) ; 饗場 篤 神戸大学, 大学院・医学系研究科, 教授 (20271116) ; 深谷 昌弘 北海道大学, 大学院・医学研究科, 助教 (10360900) ; 宮崎 太輔 北海道大学, 大学院・医学研究科, 助教 (90374230)
• Keywords: シナプス / 脳 / 発達 / 小脳 / 大脳皮質

Research Abstract

1) CNB1のfloxマウスの作成と細胞特異的Creマウスとの交配
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CNAαCNAβCNB1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。しかし、null型欠損CNB1マウスは致死的であり、神経回路の発達や機能解析には条件的CNB1欠損マウスが不可欠である。平成19年度の研究では、分担者である新潟大学の崎村教授と共同して、カルシウム結合に必須なEFハンドをコードするCNB1遺伝子の第3エクソンと第4エクソンをloxP配列で挟んだfloxマウスを作成した。さらに、崎村教授と神戸大学の饗場教授と共同して、プルキンエ細胞特異的Creマウスと大脳皮質特異的Creマウスと交配することにより、細胞特異的ノックアウトマウスの作製に成功した。今後、このノックアウトマウスを使って、小脳と大脳皮質におけるシナプス回路における分子機能を追求する。
2) CaMKIIα,CaMKIIβ,CNB1の特異抗体作成
CaMKIIおよびカルシニューリンの脳内分布解析に必要なポリクローナル抗体作成を行った。イムノブロットで検討した結果、CaMKIIβ、CNB1、CNAαCNAβ.に対する特異抗体を得ることができた。今後、これを用いて小脳と大脳皮質でのカルシニューリンの分布を検討する。

Research Products

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• [Journal Article] Subcellular arrangement of molecules for 2-arachidonoyl-glycerol-mediated retrograde signaling and its physiological contribution to synaptic modulation in the striatum. (2007)
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• [Journal Article] Expression of the vesicular glutamate trasporters-1 and -2 in adult mouse dorsal root ganglia and spinal cord and their regulation by nerve injury. (2007)
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• [Journal Article] CD3 and IgG Fc receptor regulate cerebellar functions. (2007)
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• [Journal Article] Evidence against GABA release from glutamatergic mossy fiber terminals in the developing hippocampus. (2007)
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• [Journal Article] Identification of the sites of 2-arachidonoylglycerol synthesis and action imply retrograde endocannabinoid signaling at both GABAergic and glutamatergic synapses in the ventral tegmental area. (2007)
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• [Journal Article] Genetic ablation of NMDA receptor subunit NR3B in mouse reveals motoneuronal and non-motoneuronal phenotypes. (2007)
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• [Journal Article] Involvement of nitric ocide in depolarization-induced suppression of inhibition in hippocampal pyramidal cells. (2007)
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• [Journal Article] Aberrant membranes and double-membrane structures accumulate in the axons of Atg5-null Purkinje cells before neuronal death. (2007)
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• [Journal Article] Altered features in mutant cerebellar Purkinje cells lacking junctophilins. (2007)
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• [Journal Article] RGS2 modulates GABAB receptor-GIRK channel coupling in dopamine neurons of the ventral tegmental area. (2007)
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• [Journal Article] Postsynaptic GABAB receptor signalling enhances LTD in mouse cerebellar Purkinje cells. (2007)
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• [Journal Article] IQ-ArfGEF/BRAG1 is a guanine nucleotide exchange factor for Arf6 that interacts and localizes with PSD-95 at excitatory synapses.
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• [Journal Article] Down-regulation of the CB1 cannabinoid receptor and related molecular elements of the endocannabinoid system in epileptic human hippocampus.
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• [Journal Article] Enzymatic machinery for endocannabinoid biosynthesis associated with calcium stores in glutamatergic axon terminals.
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• [Journal Article] Effects of FAK ablation on cerebellar foliation, Bergmann glia and climbing fiber territory on Purkinje cells.
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• [Presentation] 登上線維プルキンエ細胞投射系による単一支配化への分子機構 (2008)
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  • Organizer: 第113回日本解剖学会シンポジウム
  • Place of Presentation: 大分県・大分大学
  • Year and Date: 2008-03-29
• [Presentation] Selective localization of NR3A subunit at climbing fiber-interneuron synapses in the cerebellum. (2007)
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• [Presentation] 小脳における登上線維-介在ニューロン間シナプス選択的に発現するNMDA 受容体 NR3A サブユニットの分子解剖学的解析 (2007)
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  • Organizer: 第53回東北・北海道連合支部学術集会
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• [Presentation] バーグマングリアに発現するAMPA型グルタミン酸受容体の輸送様式 (2007)
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  • Organizer: Neuro 2007 第30回日本神経科学大会
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  • Year and Date: 20070910-12
• [Presentation] GLAST分子欠損マウスにおけるバーグマングリアの形態分化異常および形態学的登上線維多重支配 (2007)
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  • Organizer: Neuro 2007 第30回日本神経科学大会
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  • Year and Date: 20070910-12
• [Presentation] M1ムスカリン性アセチルコリン受容体は終脳の投射ニューコンに豊富に発現している (2007)
  • Author(s): 山崎美和子、飯塚尭、渡辺雅彦
  • Organizer: Neuro 2007 第30回日本神経科学大会
  • Place of Presentation: 横浜・パシフィコ横浜
  • Year and Date: 20070910-12
• [Presentation] マウス終脳の特定の介在神経におけるPKC&alphaおよびPKC&gammaの細胞発現パターンの解析 (2007)
  • Author(s): 三浦会里子, 窪田圭志, 渡辺雅彦
  • Organizer: Neuro 2007 第30回日本神経科学大会
  • Place of Presentation: 横浜・パシフィコ横浜
  • Year and Date: 20070910-12
• [Presentation] 線条体における2-アラキドノイルグリセロールを介した逆行性シグナル伝達に関与する分子の細胞内配置 (2007)
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  • Organizer: Neuro 2007 第30回日本神経科学大会
  • Place of Presentation: 横浜・パシフィコ横浜
  • Year and Date: 20070910-12
• [Presentation] Subcellular arrangement of molecules for 2-arachidonoyl-glycerol-mediated retrograde signaling in the striatum. (2007)
  • Author(s): Motokazu, Uchigashima., Madoka, Narushima., Masahiro, Fukaya., Istvan, Katona., Masanobu, Kano., Masahiko, Watanabe.
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• [Presentation] マウス線条体における内因性カンナビノイド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の分子機構 (2007)
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  • Year and Date: 2007-12-01
• [Presentation] 神経系における内在性カンナビノイドの分子解剖学 (2007)
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  • Organizer: 日本麻酔科学会北海道東北支部第55回北海道麻酔科学会
  • Place of Presentation: 札幌・北海道大学
  • Year and Date: 2007-09-08
• [Presentation] 神経系における内在性カンナビノイドの合成・分解とその生理作用 (2007)
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