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Roles of protein kinase C (PKC) in the formation of neural plasticity. -Investigation using GFP-tagged PKC

Research Project

Project/Area Number 14580743
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Neurochemistry/Neuropharmacology
Research InstitutionHIROSHIMA UNIVERSITY

Principal Investigator

SAKAI Norio  Hiroshima University, Graduate School of Biomedical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (70263407)

Co-Investigator(Kenkyū-buntansha) SEKI Takahiro  Hiroshima University, Graduate School of Biomedical Sciences, Research Associate, 大学院・医歯薬学総合研究科, 助手 (50335650)
AMANO Taku  Hiroshima University, Graduate School of Biomedical Sciences, Assistant Professor, 大学院・医歯薬学総合研究科, 講師 (10294547)
MATSUBAYASHI Hiroaki  Hiroshima University, Graduate School of Biomedical Sciences, Associate Professor, 大学院・医歯薬学総合研究科, 助教授 (60165850)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywordsphosphorylation / protein kinase C (PKC) / PDK-1 / transgenic mice / propagation of neural excitability / 蛋白質リン酸化 / 神経可塑性 / 小脳プルキンエ細胞 / ライブイメージング
Research Abstract

1) Development of inducible and brain region specific gamma-PKC-GFP transgenic mice and examination of PKC translocation in neurons
We have developed the tet-regulated and brain region specific PKC-GFP transgenic mice. In these mice, the expression of gamma-PKC-GFP was seen in striatum and cerebellar Purkinje cells when transgene was driven under the control of neuron specific enolase (NSE) promoter. Whereas, when the target gene was driven under the control of calcium calmodulin kinase II (CaMKII) promoter, gamma-PKC-GFP expression was observed in forebrain including hippocampus, cerebral cortex, striatum and olfactory bulb. We prepared the cerebellar brain slices and observed the translocation of gamma-PKC-GFP in vivo and living states. The translocation of gamma-PKC-GFP was elicited by agonists of metabotropic glutamate receptors (mGluR) or local electronic stimulation. In case of electronic stimulation, gamma-PKCGFP was propagated along the dendritic shaft of Purkinje cells. Our transgenic mice would be a beneficial tool for investigating the propagation of neural excitability at protein levels
2) PKC oscillations
It has bee known that activation of mGluRS results in the generation of Ca^<2+> oscillation. We found that PKC was oscillatory translocated, concomitantly with intracellular Ca^<2+>. Phosphorylation of threonine residue at 840 (T840) was crucial for the regulation of Ca^<2+> and PKC oscillation
3) Role of PDK1 (phosphoinositide-dependent kinase 1) in PKC translocation
PDK1 is a kinase that phosphorylates the threonine residues of activation loop, found in AGC kinase family, including PKC. To elucidate the role of PDK1 in PKC translocation, we constructed mutant PKC whose threonine residue in activation loop was substituted to alanine. We expressed this mutant PKC in cells and examined its translocation. We found that PDK1-insesitive PKC mutant was retained at the plasma membrane longer than wild PKC, indicating that PDK1 affect PKC targeting mechanism

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (57 results)

All Other

All Publications (57 results)

  • [Publications] Kashiwagi, K.: "Importance of C1B domain for lipid messenger-induced targeting of PKC."The Journal of Biological Chemistry. 277. 18037-18045 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakai, N.: "Inducible and brain region specific CREB transgenic mice."Molecular Pharmacology. 61. 1453-1464 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nishizaki, T.: "A new neuromodulator pathway with a glial contribution mediated via A2a adrenergic receptors."Glia. 39. 133-147 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakai, K.: "The silencer activity of the novel human serotonin transporter linked polymorphic region."Neuroscience Letters. 327. 13-16 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Seki, T.: "Adenoviral gene transfer of aspartoacylase into the tremor rat, a genetic model of epilepsy, as a trial of gene therapy for inherited epileptic disorder."Neuroscience Letters. 328. 249-252 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Goerke, A.: "Induction of apoptosis by Protein kinase Cδ is independent of its kinase activity."The Journal of Biological Chemistry. 277. 32054-32062 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kodama, N.: "Effects of D-Fraction, a polysaccharide from Grifola frondosa on tumor growth involve activation of NK cells."Biological & Pharmacological Bulletin. 25. 1647-1650 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Irie, N.: "Subtype- and species-specific knockdown of PKC using short interfering RNA."Biochemical and Biophysical Research Communications. 298. 738-743 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Newton, S.S.: "Inhibition of cAMP response element-binding protein or dynorphin in the nucleus accumbens produces an antidepressant-like effect."Journal of Neuroscience. 24. 10883-10890 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kiura, Y.: "Involvement of Ca^<2+> channels in abnormal excitability of hippocampal CA3 pyramidal cells in Noda epileptic rats."Journal of Pharmacological Sciences. 91. 137-144 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakai, K.: "Novel variants of murine serotonin transporter mRNA and the promoter activity of its upstream site."Neuroscience Letters. 342. 175-178 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Amano, T.: "Repeated administration of methamphetamine causes hypersensitivity of D_2 receptor in rat ventral tegmental area."Neuroscience Letters. 347. 89-92 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shiwa, T.: "Perospirone, a novel antipsychotic agent, hyperpolarizes rat dorsal raphe neurons via 5-HT_<1A> receptor."Journal of Pharmacological Sciences. 93. 114-117 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Matsubayashi, H.: "Electrophysiological characterization of nicotine-induced excitation of dopaminergic neurons in the rat substantia nigra."Journal of Pharmacological Sciences. 93. 143-148 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Matsubayashi, H.: "Postsynaptic α4β2 and α7 type nicotinic acetylcholine receptors (nAChRs) contribute to the local and endogenous acetylcholine-mediated synaptic transmissions in nigral dopaminergic neurons."Brain Research. 1005. 1-8 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Uchino, M: "Isoform-specific phosphorylation of mGluR5 by PKC blocks Ca^<2+> oscillation and oscillatory translocation of Ca^<2+>-dependent PKC."The Journal of Biological Chemistry. 279. 2254-2261 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kajimoto, T.: "Ceramide-induced apoptosis by translocation, phosphorylation and activation of protein kinase Cδ at Golgi complex."The Journal of Biological Chemistry. 279. 12668-12676 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Bhattacharjee, A.K.: "In vivo transgene expression using an adenoviral tetracycline-regulated system with neuron-specific enolase promoter."Biochemical and Biophysical Research Communications. 317. 1144-1148 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yagi, K.: "Phospholipase A2 products retain a neuron specific γ isoform of PKC on the plasma membrane through the C1 domain-A molecular mechanism for sustained enzyme activity."Neurochemistry International. 45. 39-47 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Seki, T.: "Adenoviral gene transfer of aspartoacylase ameliorates tonic convulsions of spontaneously epileptic rats."Neurochemistry International. 45. 171-178 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kashiwagi, K.: "Importance of C1B domain for lipid messenger-induced targeting of PKC"J.Biol.Chem.. 277. 18037-18045 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakai, N.: "Inducible and brain region specific CREB transgenic mice"Mol.Pharmacol.. 61. 1453-1464 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nishizaki, T.: "A new neuromodulator pathway with a glial contribution mediated via A2a adrenergic receptors"Glia. 39. 133-147 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakai, K.: "The silencer activity of the novel human serotonin transporter linked polymorphic region"Neurosci.Lett.. 327. 13-16 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Seki, T.: "Adenoviral gene transfer of aspartoacylase into the tremor rat, a genetic model of epilepsy, as a trial of gene therapy for inherited epileptic disorder"Neurosci Lett.. 328. 249-252 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Goerke, A.: "Induction of apoptosis by Protein kinase Cδ is independent of its kinase activity"J.Biol.Chem.. 277. 32054-32062 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kodama, N.: "Effects of D-Fraction, a polysaccharide from Grifola frondosa on tumor growth involve activation of NK cells"Biol.Pharm.Bull.. 25. 1647-1650 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Irie, N.: "Subtype-and species-specific knockdown of PKC using short interfering RNA"Biochem.Biophys.Res.Commun.. 298. 738-743 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Newton, S.S.: "Inhibition of cAMP response element-binding protein or dynorphin in the nucleus accumbens produces an antidepressant-like effect"J.Neurosci.. 24. 10883-10890 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kiura, Y: "Involvement of Ca^<2+> channels in abnormal excitability of hippocampal CA3 pyramidal cells in Noda epileptic rats"J.Pharmacol.Sci.. 91. 137-144 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakai, K.: "Novel variants of marine serotonin transporter mRNA and the promoter activity of its upstream site"Neurosci.Lett.. 342. 175-178 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Amano, T.: "Repeated administration of methamphetamine causes hypersensitivity of D_2 receptor in rat ventral tegmental area"Neurosci.Lett.. 347. 89-92 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shiwa, T: "Perospirone, a novel antipsychotic agent, hyperpolarizes rat dorsal raphe neurons via 5-HT_<1A> receptor"J.Pharmacol.Sci.. 93. 114-117 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Matsubayashi, H.: "Electrophysiological characterization of nicotine-induced excitation of dopaminergic neurons in the rat substantia nigra"J.Pharmacol.Sci.. 93. 143-148 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Matsubayashi, H.: "Postsynaptic α4β2 and α7 type nicotinic acetylcholine receptors (nAChRs) contribute to the local and endogenous acetylcholine-mediated synaptic transmissions in nigral dopaminergic neurons"Brain Res.. 1005. 1-8 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Uchino, M.: "Isoform-specific phosphorylation of mGluR5 by PKC blocks Ca^<2+> oscillation and oscillatory translocation of Ca^<2+>-dependent PKC"J.Biol.Chem.. 279. 2254-2261 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kajimoto, T.: "Ceramide-induced apoptosis by translocation, phosphorylation and activation of protein kinase Cdelta at GoIgi complex"J.Biol.Chem.. 279. 12668-12676 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Bhattacharjee, A. K.: "In vivo transgene expression using an adenoviral tetracycline-regulated system with neuron-specific enolase promoter"Biochem.Biophys.Res.Commun.. 317. 1144-1148 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yagi, K.: "Phospholipase A2 products retain a neuron specific γ isoform of PKC on the plasma membrane through the C1 domain -A molecular mechanism for sustained enzyme activity"Neurochem.Int.. 45. 39-47 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Seki, T.: "Adenoviral gene transfer of aspartoacylase ameliorates tonic convulsions of spontaneously epileptic rats"Neurochem.Int.. 45. 171-178 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shiwa, T.: "Perospirone, a novel antipsychotic agent, hyperpolarizes rat dorsal raphe neurons via 5-HT_<1A> receptor"J.Pharmacol.Sci.. 93. 114-117 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Matsubayashi, H.: "Electrophysiological characterization of nicotine-induced excitation of dopaminergic neurons in the rat substantia nigra"J.Pharmacol.Sci.. 93. 143-148 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Matsubayashi, H.: "Postsynaptic α4β2 andα7 type nicotinic acetylcholine receptors (nAChRs) contribute to the local and endogenous acetylcholine-mediated synaptic transmissions in nigral dopaminergic neurons"Brain Research. (in press). (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Uchino, M.: "Isoform-specific phosphorylation of mGluR5 by PKC blocks Ca^<2+> oscillation and oscillatory translocation of Ca^<2+>-dependent PKC"J.Biol.Chem.. (in press). (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Seki, T: "Adenoviral gene transfer of aspartoacylase ameliorates tonic convulsions of spontaneously epileptic rats"Neurochemistry Int.. (in press). (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kajimoto, T.: "Ceramide-induced apoptosis by translocation, phosphorylation and activation of protein kinase Cdelta at Golgi complex"J.Biol.Chem.. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kashiwagi, K, 他: "Importance of C1B domain for lipid messenger-induced targeting of PKC"J. Biol. Chem.. 277. 18037-18045 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sakai, N, 他: "Inducible and brain region specific CREB transgenic mice"Mol. Pharmacol.. 61. 1453-1464 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nishizaki, T, 他: "A new neuromodulator pathway with a glial contribution mediated via A2a adrenergic receptors"Glia. 39. 133-147 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sakai, K, 他: "The silencer activity of the novel human serotonin transporter linked polymorphic region"Neurosci. Lett.. 327. 13-16 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Seki, T, 他: "Adenoviral gene transfer of aspartoacylase into the tremor rat, a genetic model of epilepsy, as a trial of gene therapy for inherited epileptic disorder"Neurosci Lett. 328. 249-252 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Goerke, A, 他: "Induction of apoptosis by Protein kinase C delta is independent of its kinase activity"J. Biol. Chem.. 277. 18037-18045 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kodama, N, 他: "Effects of D-Fraction, a polysaccharide from Grifola frondosa on tumor growth involve activation of NK cells"Biol. Pharm. Bull.. 25. 1647-1650 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Irie, N., 他: "Subtype-and species-specific knockdown of PKC using short interfering RNA"Biochem. Biophys. Res. Commun.. 298. 738-743 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Newton, S.S., 他: "Inhibition of cAMP response element-binding protein or dynorphin in the nucleus accumbens produces an antidepressant-like effect"J. Neurosci. 2002. 24. 10883-10890 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kiura, Y., 他: "Involvement of Ca^<2+> channels in abnormal excitability of hippocampal CA3 pyramidal cells in Noda epileptic rats"J. Pharmacol. Sci.. 91. 137-144 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sakai, K., 他: "Novel Variant of murine serotonin transporter mRNA and the promoter activity of its upstream site"Neurosci Lett. (印刷中). (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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