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生細胞内オルガネラ膜上タンパク質の一分子可視化技術の開拓

Research Project

Project/Area Number 17659076
Research Category

Grant-in-Aid for Exploratory Research

Allocation TypeSingle-year Grants
Research Field General pharmacology
Research InstitutionNagoya City University

Principal Investigator

今泉 祐治  名古屋市立大学, 大学院薬学研究科, 教授 (60117794)

Co-Investigator(Kenkyū-buntansha) 大矢 進  名古屋市立大学, 助教授 (70275147)
山村 寿男  名古屋市立大学, 助手 (80398362)
Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2006: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2005: ¥1,900,000 (Direct Cost: ¥1,900,000)
Keywords一分子可視化 / 全反射蛍光顕微鏡 / 細胞内小器官 / 小胞体 / イオンチャネル / リアノジン受容体 / カルシウムチャネル / カルシウム活性化カリウムチャネル / カリウムチャネル / 細胞内カルシウム動態 / オルガネラ / 細胞膜
Research Abstract

全反射蛍光顕微鏡を利用して、生きた細胞内で機能している細胞内小器官、特に小胞体の膜上に存在する特定の蛋白質(特にはリアノジンCa^<2+>遊離チャネル)のリアルタイムでの機能を一分子可視化法により、解明することを試みた。さらにその技術を一般化し、その他の小胞体上の蛋白質、あるいはその他のオルガネラ(ミトコンドリアなど)の膜表面蛋白を一分子可視化し、その機能解析に新分野を切り開くための端緒とするべく検討した。
(1)膜電位固定下で平滑筋細胞膜上の1分子のまたは凝集した分子群のCa^<2+>チャネルが脱分極で開口することにより、細胞膜直下の筋小胞体からのリアノジン受容体を介したCa^<2+>遊離を可視化することに成功した(07年日本薬理学会年会発表;論文投稿準備中)。
(2)YFPでラベルされた2および3型リアノジン受容体をHEK293細胞に発現させ、リアノジン受容体開口による自発的Ca^<2+>遊離現象を一分子可視化することを試行している。
(3)CFPラベルされた大コンダクタンスCa^<2+>活性化K^+チャネルをHEK293細胞に発現させ、機能的クラスター形成の過程を一分子可視化法により明らかにした。また(2)と同時に発現させることにより両分子の機能連関の可視化を検討している。

Report

(2 results)
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • Research Products

    (17 results)

All 2007 2006 2005 Other

All Journal Article (14 results) Book (3 results)

  • [Journal Article] Methyl-β-cyclodextrin prevents Ca^<2+>-induced Ca^<2+> release in smooth muscle cells of mouse urinary bladder2007

    • Author(s)
      Hotta S, Yamamura H, Ohya S, Imaizumi Y
    • Journal Title

      J Pharmacol Sci 103(1)

      Pages: 121-126

    • NAID

      10024312258

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Voltage-sensitive oxonol dyes are novel large-conductance Ca^<2+>-activated K^+ channel activators selective for β1 and β4 but not for β2 subunits2007

    • Author(s)
      Morimoto T, Sakamoto K, Sade H, Ohya S, Muraki K, Imaizumi Y
    • Journal Title

      Mol Pharmacol 71(4)

      Pages: 1075-1088

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Novel functions of small conductance Ca^<2+>-activated K^+ channel in enhanced cell proliferation by ATP in brain endothelial cells2006

    • Author(s)
      Yamazaki D, Aoyama M, Ohya S, Muraki K, Asai K, Imaizumi Y
    • Journal Title

      J Biol Chem 281(50)

      Pages: 38430-38439

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Molecular and electrophysiological characteristics of K^+ conductance sensitive to acidic pH in aortic smooth muscle cells of WKY and SHR2006

    • Author(s)
      Kiyoshi H, Yamazaki D, Ohya S, Kitsukawa M, Muraki K, Saito SY, Ohizumi Y, Imaizumi Y
    • Journal Title

      Am J Physiol Heart Circ Physiol 291(6)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] 血管に対するチャネル開口薬の作用2006

    • Author(s)
      今泉祐治, 村木克彦
    • Journal Title

      心電図 26

      Pages: 11-19

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Two-step Ca^<2+> intracellular release underlies excitation-contraction coupling in mouse urinary bladder myocytes2006

    • Author(s)
      Morimura et al.
    • Journal Title

      American Journal of Physiology 290(2)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Molecular mechanisms for BK channel activation by a novel opener, 12,14-dichlorodehydroabietic acid2006

    • Author(s)
      Sakamoto et al.
    • Journal Title

      Journal of Pharmacology and Experimental Therapeutics 316(1)

      Pages: 144-153

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Activation of large-conductance Ca^<2+> activated K^+ channels by cannabinoids2006

    • Author(s)
      Sade et al.
    • Journal Title

      American Journal of Physiology 290(1)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Inherent pacemaker function of duodenal GIST2006

    • Author(s)
      Furuzono et al.
    • Journal Title

      European Journal of Cancer 42(2)

      Pages: 243-248

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Cardioprotective effects of estradiol include the activation of large-conductance Ca^<2+>-activated K^+ channels in cardiac mitochontdria2005

    • Author(s)
      Ohya et al.
    • Journal Title

      American Journal of Physiology 289(4)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Sulphonylurea receptors differently modulate ICC pacemaker Ca^<2+> activity and smooth muscle contractility2005

    • Author(s)
      Nakayama et al.
    • Journal Title

      Journal of Cell Science 118(18)

      Pages: 4163-4173

    • Related Report
      2005 Annual Research Report
  • [Journal Article] K^+ currents regulate the resting membrane potential, proliferation, and contractile responses in ventricular fibroblasts and myofibroblasts2005

    • Author(s)
      Chilton et al.
    • Journal Title

      American Journal of Physiology 288(6)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Ryanodine receptor type 2 deficiency changes excitation-contraction coupling and membrane potential in urinary bladder smooth muscle

    • Author(s)
      Hotta S, Kozo M, Ohya S, Muraki K, Takeshima H, Imaizumi Y
    • Journal Title

      J Physiol (in press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] CNP activates a nonselective cation current in acutely isolated rat cardiac fibroblasts via NPR-C receptor mediated signaling

    • Author(s)
      Rose RA, Hatano N, Ohya S, Imaizumi Y, Giles WR
    • Journal Title

      J Physiol (in press)

    • Related Report
      2006 Annual Research Report
  • [Book] A new insight into the function of TRPV2 in circulatory organs. TRP Ion Channel Function in Sensory Transduction and Cellular Signaling Cascades (Frontiers in Neuroscience)(Chapter28)(Wolfgang B. Liedtke, ed.)2006

    • Author(s)
      Muraki K, Shigekawa M, Imaizumi Y
    • Publisher
      CRC Press
    • Related Report
      2006 Annual Research Report
  • [Book] Ca^<2+> releasing channels in pulmonary vascular smooth muscle cells (Ion channels in the pulmonary vasculature)(edited by Yuan Jason X.-J.)2005

    • Author(s)
      Imaizumi Y, Muraki K.
    • Publisher
      Marcel Dekker Inc
    • Related Report
      2005 Annual Research Report
  • [Book] 血管平滑筋細胞イオンチャネルとリモデリング(イオンチャネル最前線update)(倉智嘉久編集)2005

    • Author(s)
      村木克彦, 今泉祐治
    • Publisher
      医歯薬出版
    • Related Report
      2005 Annual Research Report

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Published: 2005-04-01   Modified: 2016-04-21  

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