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Study on the modulation mechanism of neuronal calcuim channels

Research Project

Project/Area Number 09672222
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

KANEKO Shuji  Kyoto University Graduate School of Pharmaceutical Sciences, Associate Professor, 薬学研究科, 助教授 (60177516)

Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥2,300,000 (Direct Cost: ¥2,300,000)
KeywordsCalcium channel / Xenopus oocytes / HEK cell / opioid receptor / spinal dorsal horn / GTP-binding protein / C fiber / TRP channel / セロトニン受容体 / GABA受容体 / カルシウム チャネル / リン酸化 / 一次感覚神経 / 神経終末
Research Abstract

On the role of voltage-dependent calcium channels in the neurons, two lines of studies have been conducted : one was the experiment using Xenopus oocyte translation system in which modulation mechanism at the molecular level is to be studied ; the other was designed to investigate the modulation of calcium channel function at primary afferent nerve terminals in spinal cord slices.
The outline of results are as follows :
(1) Oocyte system : I have clarified that tonic inhibition of N-type channels by G-proteins is mediated by Gbetagamma subunit. The inhibition by Gbetagamma is competitive to that by protein kinase C, and washable by continuous intracellular perfusion with GTP-containing solution. Stimulation of coexpressed opioid receptors with agonist elicited a voltage-dependent inhibition of N-type channels, as shown in the tonic inhibition, however, the inhibition by opioids were voltage-resistant in the case of P/Q-type channels. The voltage-resistant inhibition could be isolated by applying GTPgammaS to N-type channels, and by intracellular perfusion with GTP-containing solution. I have also found two alternatively splicing variants of human N-type channels by RT-PCR.
(2) Spinal slice : EPSPs were recorded from spinal cord slices with dorsal horn attached by blind patch clamp technique. The evoked responses were categorized into two groups : one is mediated by Adelta fibers ; the other is mediated by C fibers. Repetitive high-frequency stimulation of C fiber inputs induced a slow depolarization of spinal neurons. This slow depolarization was presynaptically inhibited either by agonists for m opioid, GABA_A, GABA_B, or 5-HT_<1A> receptors. I have also clarified that N-type channels are involved in the normal excitatory neurotransmission which is mediated by glutamic acid, and that P-type channels are involved in the slow depolarization of spinal neurons.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] Kaneko,S.et al.: "Inhibition of Ca^<2+> channel curreut by μ-and κ-opioid receptors Ceexpressed in Xenopus oocytes : desensitization depeudeuce on Ca^<2+> channel α_1 subunits" Br.J.Pharmacol.121. 806-812 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kaneko,S.et al.: "Differential regulation of N-and Q-type Ca^<2+> channels by cyclic nucleotides and G-proteins" Life Sci.62. 1543-1547 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tomita,Y.et al.: "Intracellular Ca^<2+> store-operated influx of Ca^<2+> through TRP-R, a rat homolog of TRP, expressed in Xenopus oocytes" Neurosci. Lett.248. 195-198 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kinoshita,M.et al.: "Inhibitory effects of bifemelaue on brain Ca^<2+> channel subtypes expressed in Xenopus oocytes" Jpn.J.Pharmacol.78. 39-44 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kaneko,S.et al.: "Cognifive enhancers and hippocampal-long-term potentiation in vitro" Behav. Brain Res.83. 45-49 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sakurai,E.et al.: "Inhibition by [Arg^8]-vasopressin of long-term potentiation in guinea pig hippocampal slice" Jpn.J.Pharmacol. 77. 103-105 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sakurai,E.et al.: "Involvement of dendritic adhesion molecule teleucephalin in hippocampal long-term poteutiation" Neuro Report. 9. 881-886 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kaneko,S.et al.: "iCut-open' method : a new method for Xenopus oocytes that enables intracellular perfusion and stalde current recording" Folia Pharmacol. Jpn.111. 157-166 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kaneko, S.et al: "Inhibition of Ca^<2+> channel current by mu-and kappa-opioid receptors coexpressed in Xenopus oocytes : desensitization dependence on Ca^<2+> channel alpha1 subunits" Br.J.Pharmacol. 121. 806-812 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kaneko, S.et al: "Differential regulation of N-and Q-type Ca^<2+> channels by cyclic nucleotides and G-proteins" Life Sci.62. 1543-1547 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tomita, Y.et al: "Intracellular Ca^<2+> store-operated influx of Ca^<2+> through TRP-R,a rat homolog of TRP,expressed in Xenopus oocytes" Neurosci.Lett.248. 195-198 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kinoshita, M.et al: "Inhibitory effects of bifemelane on brain Ca^<2+> channel subtypes expressed in Xenopus oocytes" Jpn.J.Pharmacol.78. 39-44 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kaneko, S.et al.: "Cognitive enhancers and hippocampal long-term potentiation in vitro" Behav.Brain Res.83. 45-49 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sakurai, E.et al.: "Inhibition by [Arg_8]-vasopressin of long term potentiatino in guinea pig hippocampal slice" Jpn.J.Pharmacol.77. 103-105 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sakurai, E.et al.: "Involvement of dentritic adhesion molecule telencephalin in hippocampal long-term potentiation" Neuroreport. 9. 881-886 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kaneko, S.: "'Cut-Open'method : a new method for Xenopus oocytes that enables intracellular perfusion and stable current recording" Folia Phrmacol.Jpn.111. 157-166 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tomita,T.et al.: "Intracellular Ca^<2+> store-operated influx of Ca^<2+> through TRR-R,and homolog of TRP,expressed in Xenopus oocytes" Neuroscience Letters. 248. 195-198 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sakurai,E.et al.: "Involvement of dendritic adhesion molecule telencephalin in hippocampal long-term potentiation" Neuro Repont. 9. 881-886 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kinoshita,M.et al.: "Inhibitory effects of bifemelane on brain Ca^<2+> channel subtypes expressed in Xenopus oocytes" Japanese Journal of Pharmacology. 78. 39-44 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kaneko,S.et al.: "Methods in Enzymology-Ion Channels vol.293" Academic Press,Inc., 13 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kaneko,S.et al.: "Inhibition of Ca^<2+> channel current by μ-and k-opioid receptors coexpressed in Xenopus oocytes" Br.J.Pharmacol.121. 806-812 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kaneko,S.et al.: "Cognitive enhancers and hippocampal long-term potentiation in vitro" Behav.Brain Res.83. 45-49 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 金子 周司: "Cut-Open法-Xenopus卵母細胞で安定した膜電流の測定と細胞内潅流を同時に可能にする新しい手法" 日本薬理学雑誌. 111. 157-166 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Mizukami,K.et al.: "Characterization of slow depolarization of substantia gelatinosa neuron by dorsal root repetitive stimulation" Neurosci.Res.(印刷中). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kaneko,S.et al.: "Differential regulation of N-and Q-type Ca^<2+> channels by cyclic nucleotides and G-proteins" Life Sci.62. 1543-1547 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kaneko,S.et al: "Methods in Enzymology-Ion Channels" Academic Press,Inc.(印刷中), (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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