発達脳におけるシナプス除去と機能成熟に関与するシグナル伝達系

• Project/Area Number: 12053226
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kanazawa University
• Principal Investigator: 狩野 方伸 金沢大学, 大学院・医学系研究科, 教授 (40185963)
• Co-Investigator(Kenkyū-buntansha): 田端 俊英 金沢大学, 大学院・医学系研究科, 助手 (80303270)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥60,200,000 (Direct Cost: ¥60,200,000); Fiscal Year 2004: ¥11,600,000 (Direct Cost: ¥11,600,000); Fiscal Year 2003: ¥11,900,000 (Direct Cost: ¥11,900,000); Fiscal Year 2002: ¥11,700,000 (Direct Cost: ¥11,700,000); Fiscal Year 2001: ¥12,800,000 (Direct Cost: ¥12,800,000); Fiscal Year 2000: ¥12,200,000 (Direct Cost: ¥12,200,000)
• 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 / カルシウム / GABA / カルシウムチャネル / スライス / 培養 / 共培養 / 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の分子機構を解明することを目標とし、種々の自然発生ミュータントマウスおよび遺伝子改変マウスの解析を行ってきた。本年度は、高閾値型のP/Qタイプカルシウムチャネルを形成するα1Aサブユニットのノックアウトマウス(α1A-/-)と、抑制性伝達物質のGABA合成酵素GAD67のノックアウトマウスと野生型マウスとのヘテロマウス(GAD67+/-)を調べた。
生後3〜4週目のα1A-/-では、80%以上のプルキンエ細胞が複数の登上線維による多重支配を受けていることが電気生理学的及び形態学的解析の結果明らかになった。生後1日目からの発達の様子を電気生理学的に詳細に解析した結果、α1A-/-では、生後10日までの過剰な登上線維シナプス除去(前期シナプス除去過程)が著しく障害されていることが判明した。一方、生後10日から16日の間の除去(後期シナプス除去過程)は正常におこった。これから、P/Qタイプカルシウムチャネルを介するプルキンエ細胞へのカルシウム流入が、前期シナプス除去過程に必須であり、後期シナプス除去過程には別の機構が関与することが示唆された。
GAD67+/-では、成熟しても多重登上線維支配を受けるプルキンエ細胞の割合が野生型マウスに比べて明らかに高いという予備的結果を得た。このGAD67+/-の生後1日目からの発達を電気生理学的および形態学的に詳細に解析するとともに、もうひとつのGABA合成酵素であるGAD65のノックアウトマウスを調べ、登上線維シナプス除去と機能成熟に対する抑制性シナプス入力の役割を明らかにするのが今後の課題である。

Research Products

• [Journal Article] Phospholipase Cβ serves as a coincidence detector through its Ca^<2+> dependency for triggering endocannabinoid signal. (2005)
  • Author(s): Hashimotodani, Y.
  • Journal Title: Neuron 45 Pages: 257-268
• [Journal Article] GABAergic activation of an inwardly rectifying K^+ current in mouse cerebellar Purkinje cells. (2005)
  • Author(s): Tabata, T.
  • Journal Title: J.Physiol.(London) 563 Pages: 443-457
• [Journal Article] Two distinct classes of muscarinic action on hippocampal inhibitory synapses : M_2-mediated direct suppression and M_1/M_3-mediated indirect suppression through endocannabinoid signaling. (2004)
  • Author(s): Fukudome, Y.
  • Journal Title: Eur.J.Neurosci. 19 Pages: 2682-2692
• [Journal Article] Distinct roles of Gα_q and Gα_<11> for Purkinje cell signaling and motor behavior. (2004)
  • Author(s): Hartmann, J.
  • Journal Title: J.Neurosci. 24 Pages: 5119-5130
• [Journal Article] Altered agonist sensitivity and desensitization of neuronal mGluR1 responses in knock-in mice by a single amino acid substitution at the PKC phosphorylaion site. (2004)
  • Author(s): Sato, M.
  • Journal Title: Eur.J.Neurosci. 20 Pages: 947-955
• [Journal Article] Ca^<2+> activity at GABA_B receptors constitutively promotes metabotropic glutamate signaling in the absence of GABA. (2004)
  • Author(s): Tabata, T.
  • Journal Title: Proc.Natl.Acad.Sci.USA. 101 Pages: 16952-16957
• [Journal Article] A novel action of stargazin as an enhancer of AMPA receptor activity. (2004)
  • Author(s): Yamazaki, M.
  • Journal Title: Neurosci.Res. 50 Pages: 369-374
• [Journal Article] Signaling complex formation of phospholipase Cβ4 with mGluR1a and IP3 receptor at the perisynapse and endoplasmic reticulum in the mouse brain. (2004)
  • Author(s): Fukaya, M.
  • Journal Title: Eur.J.Neurosci. 20 Pages: 2929-2944
• [Journal Article] Retrograde modulation of synaptic transmission mediated by endogenous cannabinoids. (2004)
  • Author(s): Kano, M.
  • Journal Title: Curr.Neuropharmacol. 2 Pages: 49-57
  • NAID: 130005447340
• [Journal Article] Calcium-dependence of native metabotropic glutamate receptor signaling in central neurons. (2004)
  • Author(s): Tabata, T.
  • Journal Title: Mol.Neurobiol. 29 Pages: 261-270
• [Publications] Fukudome, Y.: "Insulin-like growth factor-I as a promoting factor for cerebellar Purkinje cell development."Eur.J.Neurosci.. 17. 2006-2016 (2003)
• [Publications] Hashimoto, K.: "Functional differentiation of multiple climbing fiber inputs during synapse elimination in the developing cerebellum."Neuron. 38. 785-796 (2003)
• [Publications] Ohno-Shosaku, T.: "Postsynaptic M_1 and M_3 receptors are responsible for the muscarinic enhancement of retrograde endocannabinoid signaling in the hippocampus."Eur.J.Neurosci.. 18. 109-116 (2003)
• [Publications] Takeda, Y.: "Impaired motor coordination in mice lacking neural recognition molecule NB-3 of the contactin/F3 subgroup."J.Neurobiol.. 56. 252-265 (2003)
• [Publications] Kitao, Y.: "ORP15O/HSP12A regulates Purkinje cell survival : A role for ER stress in cerebellar development."J.Neurosci.. 24. 1486-1496 (2004)
• [Publications] Miyazaki, T.: "P/Q-type Ca^<2+> channel α1A regulates synaptic competition on developing cerebellar Purkinje cells."J.Neurosci.. 24. 1734-1743 (2004)
• [Publications] Kano, M.: "Retrograde modulation of synaptic transmission mediated by endogenous cannabinoids."Curr.Neuropharmacol.. 2. 49-57 (2004)
• [Publications] Kano, M.: "Endocannabinoid-mediated modulation of excitatory and inhibitory synaptic transmission. In : EXCITATORY-INHIBITORY BALANCE.(T.K.Hensch, M.Fagiolini, (eds))"Kluwer Academic / Pllenum Publishers, New York/Boston/Dordrecht/London/Moscow. 11 (2003)
• [Publications] 橋本浩一: "小脳シナプスの機能発達、可塑性および機能調節におけるグルタミン酸シグナル伝達の役割(脳血管障害による「神経細胞死」の予防と治療)(川合述史/編)"クバプロ. 16 (2003)
• [Publications] 狩野方伸: "内因性カンナビノイドによる逆行性シナプス伝達 学術月報"日本学術振興会. 5 (2003)
• [Publications] 橋本浩一: "発達期小脳における登上線維シナプスの機能分化 実験医学(御子柴克彦、真鍋俊也、三浦正幸/編)"羊土社. 6 (2003)
• [Publications] 橋本浩一: "発達期小脳におけるシナプスの機能分化 蛋白質 核酸 酵素 49 No.3(増刊号/神経回路の機能発現のメカニズム)(大森治紀、渋木克栄、 野田亮、山森哲雄/編)"共立出版. 7 (2004)
• [Publications] Kano, M.: "Retrograde signaling at central synapses via endogenous cannabinoids"Mol. Psychiatry. 7. 234-235 (2002)
• [Publications] Yoshida, T.: "The cannabinoid CB1 receptor mediates retrograde signals for depolarization-induced suppression of inhibition in cerebellar Purkinje cells"J.Neurosci.. 22. 1690-1697 (2002)
• [Publications] Ohno-Shosaku, T.: "Cooperative endocannabinoid production by neuronal depolarization and group I metabotropic glutamate receptor activation"Eur.J.Neurosci.. 15. 953-961 (2002)
• [Publications] Nishi, M.: "Motor discoordination in mutant mice lacking junctophilin type 3"Biochem.Biophysic.Res.Comm.. 292. 318-324 (2002)
• [Publications] Ohno-Shosaku, T.: "Presynaptic cannabinoid sensitivity is a major determinant of depolarization-induced retrograde suppression at hippocampal synapses"J.Neurosci.. 22. 3864-3872 (2002)
• [Publications] Tabata, T.: "Extracellular calcium controls the dynamic range of neuronal metabotropic glutamate receptor responses"Mol.Cell.Neurosci.. 20. 56-68 (2002)
• [Publications] Watase, K.: "A long CAG repeat in the mouse Scal locus replicates SCA1 features and reveals the impact of mutant protein solubility on selective neuronal vulnerability"Neuron. 34. 905-919 (2002)
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• [Publications] Ichikawa, R.: "Distal extension of climbing fiber territory and multiple innervation caused by aberrant wiring to adjacent spiny branchlets in cerebellar Purkinje cells lacking glutamate receptor GluRδ"J.Neurosci.. 22. 8487-8503 (2002)
• [Publications] Kishimoto, Y.: "mGluR1 in cerebellar Purkinje cells is required for normal association of temporally contiguous stimuli in classical conditioning"Eur.J.Neurosci.. 16. 2416-2424 (2002)
• [Publications] Kakizawa, S.: "Effects of insulin-like growth factor I on climbing fiber synapse elimination during cerebellar development"Eur.J.Neurosci.. 17. 545-554 (2003)
• [Publications] Takeda, Y.: "Impaired motor coordination in mice lacking neural recognition molecule NB-3 of the contactin/F3 subgroup"J.Neurobiol.. (in press).
• [Publications] 少作隆子: "グルタミン酸受容体チャネルとシナプス可塑性"医学のあゆみ. 201. 1123-1127 (2002)
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• [Publications] T.Ohno-Shosaku: "Endogenous cannabinoids mediate retrograde signals from depolarized postsynaptic neurons to presynaptic terminals"Neuron. 29. 729-738 (2001)
• [Publications] A.W.McGee: "PSD-93 knock-out mice reveal that neuronal MAGUKs are not required for development or function of parallel fiber synapses in cerebellum"J.Neurosci. 21. 3085-3091 (2001)
• [Publications] M.Miyata: "Deficient long-term synaptic depression in the rostral cerebellum correlated with impaired motor learning in phospholipase C β4 mutant mice"Eur.J.Neurosci.. 13. 1945-1954 (2001)
• [Publications] T, Maejima: "Presynaptic in hibition caused by retrograde signal from metabotropic glutamate to cannabinoid receptors"Neuron. 31. 463-475 (2001)
• [Publications] T, Maejima: "Endogenous cannabinoid as a retrograde messenger from depolarized postsynaptic neurons to presynaptic terminals"Neurosci.Res.. 40. 205-210 (2001)
• [Publications] K.Hashimoto: "Roles of glutamate receptor δ2 subunit (GluRδ2) and metabotropic glutamate receptor subtype 1(mGluR1) in climbing fiber synapse elimination during postnatal cerebellar development"J.Neurosci.. 21. 9701-9712 (2001)
• [Publications] K.Hashimoto: "Roles of phospholipase C β4 in synapse elimination and plasticity in developing and mature cerebellum"Mol.Neurobiol.. 23. 69-82 (2001)
• [Publications] T.Tabata: "Heterogeneous intrinsic firing properties of vertebrate retinal ganglion cells"J.Neurophysiol.. 87. 30-41 (2002)
• [Publications] T.Yoshida: "The cannabinoid CB1 receptor mediates retrograde signals for depolarization-induced suppression of inhibition in cerebellar Purkinje cells"J.Neurosci.. 22. 1690-1697 (2002)
• [Publications] M.Kano: "Retrograde signaling at central synapses via endogenous cannabinoids"Mol.Psychiatry. 7(in press). (2002)
• [Publications] T.Ohno-Shosaku: "Cooperative endocannabinoid production by neuronal depolarization and group I metabotropic glutamate receptor activation"Eur. J. Neurosci.. (in press). (2002)
• [Publications] 橋本浩一: "小脳シナプス可塑性の分子機構"神経研究の進歩. 49. 249-260 (2001)
• [Publications] 橋本浩一: "小脳プルキンエ細胞におけるシナプス可塑性とカルシウム"Clinical Calcium. 11. 1432-1439 (2001)
• [Publications] 少作隆子: "脳内カンナビノイド受容体とシナプス伝達調節"脳の科学. 23. 1000-1002 (2001)
• [Publications] 橋本浩一: "発達脳におけるシナプス除去のメカニズム"医学のあゆみ. 199. 531-534 (2001)
• [Publications] 前島隆司: "カンナビノイド受容体と逆行性シグナル伝達"Clinical Neuroscience. 19. 1431 (2001)
• [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)
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