Molecular anatomical studies on the localization and functions of NMDA receptors in inhibitory neurons

• Project/Area Number: 19700315
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: Hokkaido University
• Principal Investigator: FUKAYA Masahiro Hokkaido University, 大学院医学研究科, 助教 (10360900)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,690,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥390,000); Fiscal Year 2008: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2007: ¥2,000,000 (Direct Cost: ¥2,000,000)
• Keywords: NMDA受容体 / 抑制性ニューロン / NR2D / NR3A / 大脳皮質 / 海馬 / 小脳 / 免疫電子 / 顕微鏡法 / 免疫電子顕微鏡法

Research Abstract

本研究課題では、抑制性ニューロンにおいて発現するNMDA受容体の調節サブユニットであるNR2DおよびNR3Aサブユニットに着目し、その発現細胞と細胞内局在、それらを介する神経回路、シナプス可塑性への関与の有無を明らかにすることを目的として研究を行った。
まず、マウス大脳皮質および海馬での抑制制ニューロンにおけるNMDA受容体NR2Dサブユニットに対する特異抗体を用いた局在解析を蛍光多重免疫染色法や免疫電子顕微鏡法を用いて行った。
その結果、大脳皮質のNR2Dサブユニットは、parvalbumin陽性介在抑制性ニューロンの非対称性シナプスの後シナプス部位に局在していた。
また、これらの前シナプスはVGluT1, VGluT2陽性であることから大脳皮質-大脳皮質間と視床-大脳皮質間のシナプス両方に発現することが明らかとなった。
さらに、海馬においてもparvalbumin陽性介在抑制性ニューロンに発現していた。
これらの結果から、NR2Dサブユニットは一部の介在ニューロンの興奮性シナプス後部で機能していることが示唆された。
一方、小脳におけるNR3Aサブユニットに対する特異抗体を用いた局在解析を行ったところ、小脳分子層の介在抑制性ニューロンであるStellate細胞およびBasket細胞に発現し、免疫電子顕微鏡解析から登上線維終末に接する介在ニューロンの樹上突起に局在することが明らかとなった。
また、この部分は興奮性シナプスの構造は認められず対称性シナプス様の構造を有していたが、シナプス前終末でのactive zoneに見られるシナプス小胞の集積が認められず、シナプス結合ではないと考えられた。さらに大脳皮質や海馬においても同様の局在が介在ニューロンで認められた。
このような結合領域にNMDA受容体が局在することはこれまでに報告がなく、NMDA受容体が全く新しい機能を有する可能性が示唆された。

Research Products

• [Journal Article] Ablation of NMDA Receptors Enhances the Excitability of Hippocampal CA3 Neurons (2009)
  • Author(s): Fukushima F, Nakao K, Shinoe T, Fukaya M, Muramatsu S, Sakimura K, Kataoka H, Mori H, Watanabe M, Manabe T, Masayoshi M
  • Journal Title: PLoS ONE 4(1)
  • Peer Reviewed
• [Journal Article] Ablation of NMDA receptors enhances the excitability of hippocampal CA3 neurons (2009)
  • Author(s): Fukushima, F., Nakao, K., Shinoe, T., Fukava, M., Muramatsu S, Sakimura, K., Kataoka, H., Mori, H., Watanabe, M., Manabe, T., Mishina, M.
  • Journal Title: ProS ONE 4
  • Peer Reviewed
• [Journal Article] Diacylglycerol kinase beta accumulates on the perisynaptic site of medium spiny neurons in the striatum (2008)
  • Author(s): Hozumi Y, Fukaya M, Adachi N, Saito N, Otani K, Kondo H, Watanabe M, Goto K
  • Journal Title: Eur J Neurosci 28(12) Pages: 2409-2422
  • Peer Reviewed
• [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)
  • Author(s): Tachikawa M, Fujinawa J, Takahashi M, Kasai Y, Fukaya M, Sakai K, Yamazaki M, Tomi M, Watanabe M, Sakimura K, Terasaki T, Hosoya K
  • Journal Title: J Neurochem 107(3) Pages: 768-778
  • Peer Reviewed
• [Journal Article] Synapse formation and clustering of neuroligin-2 in the absence of GABAA receptors (2008)
  • Author(s): Patrizi A, Scelfo B, Viltono L, Briatore F, Fukaya M, Watanabe M, Strata P, Varoqueaux F, Brose N, Fritschy JM, Sassoe-Pognetto M
  • Journal Title: Proc Natl Acad Sci U S A 105(35) Pages: 13151-13156
  • Peer Reviewed
• [Journal Article] Predominant expression of phospholipase Cβ1 in telencephalic principal neurons and cerebellar interneurons, and its close allocation with related signaling molecules in somatodendritic neuronal elements (2008)
  • Author(s): Fukaya M, Uchigashima M, Nomura S, Hasegawa U, Kikuchi H, Watanabe M
  • Journal Title: Eur J Neurosci 28(9) Pages: 1744-1759
  • Peer Reviewed
• [Journal Article] Enhancement of both long-term depression induction and optokinetic response adaptation in mice lacking delphilin (2008)
  • Author(s): Takeuchi T, Ohtsuki G, Yoshida T, Fukaya M, Wainai T, Yamashita M, Yamazaki Y, Mori H, Sakimura K, Kawamoto S, Watanabe M, Hirano T, Mishina M
  • Journal Title: PLoS ONE 3(5)
  • Peer Reviewed
• [Journal Article] Use-dependent amplification of presynaptic Ca2+ signaling by axonal ryanodine receptors at the hippocampal mossy fiber synapse (2008)
  • Author(s): Shimizu H, Fukaya M, Yamasaki M, Watanabe M, Manabe T, Kamiya H
  • Journal Title: Proc Natl Acad Sci U S A 105(33) Pages: 11998-12003
  • Peer Reviewed
• [Journal Article] Cloning and characterization of E-dlg, a novel splice variant of mouse SAP97 binds preferentially to SAP102 (2008)
  • Author(s): Mao P, Tao Y, Fukaya M, Li D, Tao F, Levine CF, Watanabe M, Johns RA
  • Journal Title: IUBMB Life 60(10) Pages: 684-692
  • Peer Reviewed
• [Journal Article] Glutamate transporters regulate lesion-induced plasticity in the developing somatosensory cortex (2008)
  • Author(s): Takasaki C, Okada R, Mitani A, Fukaya M, Yamasaki M, Fujihara Y, Shirawawa T, Tanaka K, Watanabe M
  • Journal Title: J Neurosci 28(19) Pages: 4995-5006
  • NAID: 120005183168
  • Peer Reviewed
• [Journal Article] Functional contributions of synaptically localized NR2B subunits of the NMDA receptor to synaptic transmission and long-term potentiation in the adult mouse CNS (2008)
  • Author(s): Miwa H, Fukaya M, Watabe AM, Watanabe M, Manabe T
  • Journal Title: J Physiol 586(10) Pages: 2539-2550
  • Peer Reviewed
• [Journal Article] Effects of FAK ablation on cerebellar foliation, Bergmann glia positioning and climbing fiber territory on Purkinje cells (2008)
  • Author(s): Watanabe F, Miyazaki T, Takeuchi T, Fukaya M, Nomura T, Noguchi S, Mori H, Sakimura K, Watanabe M, Mishina M
  • Journal Title: Eur J Neurosci 27(4) Pages: 836-854
  • Peer Reviewed
• [Journal Article] Q-ArfGEF/BRAG1 is a guanine nucleotide exchange factor for Arf6 that interacts with PSD-95 at postsynaptic density of excitatory synapses (2008)
  • Author(s): Sakagami H, Sanda M, Fukaya M, Miyazaki T, Sukegawa J, Yanagisawa T, Suzuki T, Fukunaga K, Watanabe M, Kondo HI
  • Journal Title: Neurosci Res 60(2) Pages: 199-212
  • Peer Reviewed
• [Journal Article] IQ-ArfGEF/BRAG1 is a guanine nucleotide exchange factor for Arf6 that interacts and localizes with PSD-95 at excitatory synapses (2008)
  • Author(s): Sakagami H, Sanda M, Fukaya M, Miyazaki T, Sukegawa J, Yanagisawa T, Suzuki T, Fukunaga K, Watanabe M, Kondo K
  • Journal Title: Neurosci Res 60 Pages: 199-212
  • Peer Reviewed
• [Journal Article] Effects of FAK ablation on cerebellar foliation, Bergmann glia and climbing fiber territory on Purkinje cells (2008)
  • Author(s): Watanabe F, Miyazaki T, Takeuchi T, Fukaya M, Nomura T, Noguchi S, Mori H, Sakimura K, Watanabe M, Mishina M
  • Journal Title: Eur. J. Neurosci 27 Pages: 836-854
  • Peer Reviewed
• [Journal Article] Functional contributions of synaptically localized NR2B subunits of the NMDA receptor to synaptic transmission and long-term potentiation in the adultmouse CNS (2008)
  • Author(s): Miwa H, Fukava M, Watabe AM, Watanabe M, Manabe T
  • Journal Title: J. Physiol 586(10) Pages: 2539-2550
  • Peer Reviewed
• [Journal Article] Glutamate transporters regulate lesion-induced period plasticity in the developing somatosensory cortex (2008)
  • Author(s): Takasaki C, Okada R, Mitani A, Fukava, M, Yamasaki M, Fujihara Y, ShirakawaT. Tanaka K. Watanabe M
  • Journal Title: J. Neurosci 28 Pages: 4995-5006
  • Peer Reviewed
• [Journal Article] Synapse formation and clustering of neuroligin-2 in the absence of GABAA receptors (2008)
  • Author(s): Patrizi A, Scelfo B, Viltono L, Briatore F, Fukaya M, Watanabe M, Strata P, Varoqueaux F, Brose N, Fritschy JM, Sassoe-Poonetto M
  • Journal Title: Proc. Natl. Acad. Sci. USA 105(35) Pages: 13151-13156
  • Peer Reviewed
• [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 SAP102 (2008)
  • Author(s): Mao P, Tao Y-X, Fukava M, Li D, Tao T, Watanabe M, Johns RA
  • Journal Title: IUBMB(International Union of Biochemistry and Molecular Biology)Life 60 Pages: 684-692
  • Peer Reviewed
• [Journal Article] Enhanced cerebellar LTD induction and OKR adaptation in mice lacking delphilin (2008)
  • Author(s): Takeuchi, T., Ohtsuki, G., Yoshida, T., Fukava, M., Wainai, T., Yamashita M, Yamazaki Y, Mori, M., Sakimura, K., Kawamoto, S., Watanabe, M., Hirano T Mishrina. M.
  • Journal Title: PLoS One(on-line journal) 3(5)
  • Peer Reviewed
• [Journal Article] Use-dependent amplification of presynaptic Ca^<2+> signaling by axonal ryanodine receptors at the hippocampal mossy fiber synapse (2008)
  • Author(s): Shimizu H, Fukava M, Yamazaki M, Watanabe M. Manabe T, Kamiva H
  • Journal Title: Proc. Natl. Acad. Sci. USA 105 Pages: 11998-12003
  • Peer Reviewed
• [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)
  • Author(s): Tachikawa M, Fujinawa J, Takahashi M, Kasai Y, Fukava M, Sakai K, Yamazaki M, Tomi M, Watanabe M, Sakimura K, Terasaki T, Hosoya K
  • Journal Title: J. Neurochem 107 Pages: 768-778
  • Peer Reviewed
• [Journal Article] Predominant expression of phospholipase CR1 in telencephalic principal neurons and cerebellar interneurons, and its close allocation with related signaling molecules in somatodendritic neuronal elements (2008)
  • Author(s): Fukava M, Uchigashima M, Nomura S, Hasegawa Y, Kikuchi H, Watanabe M
  • Journal Title: Eur J Neurosci 28 Pages: 1744-1759
  • Peer Reviewed
• [Journal Article] Diacylglycerol kinase β accumulates on the perisynaptic site of medium spiny neurons in the striatum (2008)
  • Author(s): Hozumi Y, Fukava M, Adachi N, Saito N, Otani K, Kondo H, Watanabe M, Goto K
  • Journal Title: Eur J Neurosci 28 Pages: 2409-2422
  • Peer Reviewed
• [Journal Article] Axonal motility and its modulation by activity are branch-type specific in the intact adult cerebellum (2007)
  • Author(s): Nishiyama H, Fukaya M, Watanabe M, Linden D
  • Journal Title: Neuron 56(3)
  • Peer Reviewed
• [Journal Article] Genetic ablation of NMDA receptor subunit NR3B in mouse reveals motoneuronal and non-motoneuronal phenotypes (2007)
  • Author(s): Niemann S, Kanki H, Fukui Y, Takao K, Fukaya M, Hynynen MN, Churchill MJ, Shefner JM, Bronson RT, Watanabe M, Brown RHJr, Miyakawa T, Itohara S, Hayash T
  • Journal Title: Eur J Neurosci 26(6) Pages: 1407-1420
  • Peer Reviewed
• [Journal Article] Evidence against GABA release from glutamatergic mossy fiber terminals in the developing hippocampus (2007)
  • Author(s): Uchigashima M, Fukaya M, Watanabe M, Kamiya H
  • Journal Title: J Neurosci 27(30) Pages: 8088-8100
  • Peer Reviewed
• [Journal Article] Subcellular arrangement of molecules for 2-arachidonoyl-glycerol-mediated retrograde signaling and its physiological contribution to synaptic modulation in the striatum (2007)
  • Author(s): Uchigashima M, Narushima M, Fukaya M, Katona I, Kano M, Watanabe M
  • Journal Title: J Neurosci 27(14) Pages: 3663-3676
  • NAID: 120005183167
  • Peer Reviewed
• [Journal Article] Junctophilin-mediated channel crosstalk essential for cerebellar synaptic plasticity (2007)
  • Author(s): Kakizawa S, Kishimoto Y, Hashimoto K, Miyazaki T, Furutani1 K, Shimizu H, Fukaya M, Nishi M, Sakagami H, Ikeda A, Kondo H, Kano M, Watanabe M, Iino M, Takeshima H
  • Journal Title: EMBO J 26(7) Pages: 1924-1933
  • Peer Reviewed
• [Journal Article] Vesicular release of glutamate from unmyelinated axons in white matter (2007)
  • Author(s): Ziskin J L, Nishiyama A, Rubio M, Fukaya M, Bergles D E
  • Journal Title: Nat Neurosci 10(3) Pages: 321-330
  • Peer Reviewed
• [Journal Article] Phospholipase Cβ3 is distributed in both somatodendriteic and axonal compartments and localized around perisynapse and smooth endoplasmic reticulum in mouse Purkinje cell subsets (2007)
  • Author(s): Nomura S, Fukaya M, Tsujioka T, Wu D, Watanabe M
  • Journal Title: Eur J Neurosci 25(3) Pages: 659-672
  • Peer Reviewed
• [Journal Article] Tonic enhancement of endocannabinoid-mediated retrograde suppression of inhibition by cholinergic interneuron activity in the striatum (2007)
  • Author(s): Narushima M, Uchigashima M, Fukaya M, Matsui M, Manabe T, Hashimoto K, Watanabe M, Kano M
  • Journal Title: J Neurosci 27 Pages: 496-506
  • Peer Reviewed
• [Journal Article] Axonal motility and its modulation by activity are branch-type specific in the intact adult cerebellum (2007)
  • Author(s): Nishiyama H, Fukaya M, Watanabe M, Linden D
  • Journal Title: Neuron 56 Pages: 472-487
  • Peer Reviewed
• [Journal Article] Genetic ablation of NMDA receptor subunit NR3B in mouse reveals motoneuronal and non-motoneuronal phenotypes (2007)
  • Author(s): Niemann S, Kanki H, Fukui Y, Takao K, Fukaya M, Hynynen MN, Churchill MJ, Shefner JM, Bronson RT, Watanabe M, Brown RHJr, Miyakawa T, Itohara S, Hayashi Y
  • Journal Title: European Journal of Neuroscience 26 Pages: 1407-1420
  • Peer Reviewed
• [Journal Article] Evidence against GABA release from glutamatergic mossy fiber terminals in the developing hippocampus (2007)
  • Author(s): Uchigashima M, Fukaya M, Watanabe M, Kamiya H
  • Journal Title: The Journal of Neuroscience 27 Pages: 8088-8100
  • Peer Reviewed
• [Journal Article] Subcellular arrangement of molecules for 2-arachidonoyl-glycerol-mediated retrograde signaling and its physiological contribution to synaptic modulation in the striatum (2007)
  • Author(s): Uchigashima M, Narushima M, Fukaya M, Katona I, Kano M, Watanabe M
  • Journal Title: The Journal of Neuroscience 27 Pages: 3663-3676
  • NAID: 120005183167
  • Peer Reviewed
• [Journal Article] Junctophilin-mediated channel crosstalk essential for cerebellar synaptic plasticity (2007)
  • Author(s): Kakizawa S, Kishimoto Y, Hashimoto K, Miyazaki T, Furutanil K, Shimizu H, Fukaya M, Nishi M, Sakagami H, Ikeda A, Kondo H, Kano M, Watanabe M, Iino M, Takeshima H
  • Journal Title: EMBO Journal 26 Pages: 1924-1933
  • Peer Reviewed
• [Journal Article] Vesicular release of glutamate from unmyelinated axons in white matter (2007)
  • Author(s): Ziskin J L, Nishiyama A, Rubio M, Fukaya M, Bergles DE
  • Journal Title: Nature Neuroscience 10 Pages: 321-330
  • Peer Reviewed
• [Journal Article] Phospholipase Cb3 is distributed in both somatodendriteic and axonal compartments and localized around perisynapse and smooth endoplasmic reticulum in mouse Purkinje cell subsets (2007)
  • Author(s): Nomura S, Fukaya M, Tsujioka T, Wu D, Watanabe M
  • Journal Title: European Journal of Neuroscience 25 Pages: 659-672
  • Peer Reviewed
• [Journal Article] Tonic enhancement of endocannabinoid-mediated retrograde suppression of inhibition by cholinergic interneuron activity in the striatum (2007)
  • Author(s): Narushima M, Uchigashima M, Fukaya M, Matsui M, Manabe T, Hashimoto K, Watanabe M, Kano M
  • Journal Title: The Journal of Neuroscience 27 Pages: 496-506
  • Peer Reviewed
• [Presentation] Selective localization of NR3A subunit at climbing fiber-interneuron synapses in the cerebellum (2007)
  • Author(s): Masahiro Fukaya, Masahiko Watanabe
  • Organizer: Society for Neuroscience 37th Annual Meeting
  • Place of Presentation: San Diego
• [Presentation] The mode of AMPA-type glutamate receptor trafficking in Bergmann glia. (2007)
  • Author(s): 深谷昌弘、渡辺雅彦
  • Organizer: 第30回 日本神経科学大会
  • Place of Presentation: 横浜
• [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)
  • Author(s): 深谷昌弘、渡辺雅彦
  • Organizer: 日本解剖学会第53回東北・北海道連合支部学術集会
  • Place of Presentation: 札幌
• [Presentation] Selective localization of NR3A subunit at climbing fiber-interneuron synapses in the cerebellum (2007)
  • Author(s): Masahiro Fukaya, and Masahiko Watanabe
  • Organizer: Society for Neuroscience 37th Annual Meeting
  • Place of Presentation: サンディエゴコンベンションセンター・San Diego, USA
• [Presentation] The mode of AMPA-type glutamate receptor trafficking in Bergmannglia (2007)
  • Author(s): 深谷昌弘、渡辺雅彦
  • Organizer: 第30回 日本神経科学大会
  • Place of Presentation: 横浜・パシフィコ横浜
• [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)
  • Author(s): 深谷昌弘、渡辺雅彦
  • Organizer: 日本解剖学会第53回東北・北海道連合支部学術集会
  • Place of Presentation: 札幌・北海道医療大学