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Molecular and Electropharmacological Analysis of Capacitative Ca^<2+> Entry channels of Vascular Endothelial Cells

Research Project

Project/Area Number 09670087
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General pharmacology
Research InstitutionAkita University

Principal Investigator

IIJIMA Toshihiko  Akita Univ.Sch.Med, Dept.Pharmacol, Professor, 医学部, 教授 (30004724)

Co-Investigator(Kenkyū-buntansha) ONO Kyoichi  Akita Univ.Sch.Med, Dept.Pharmacol, Assoc.Professor, 医学部, 助教授 (70185635)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsVascular Endothel / Intracellular Ca^<2+> concentrarion / Fura-2 / Capacitative Ca^<2+> Entry Chanell / Patch Clamp / Cl^- Current / Fluid-stream / trp / Cl^-電流 / trp / Fura‐2
Research Abstract

In cultured human aortic endothelial cells (HAECs) histamine (1-100 muM) produced a biphasic response in [Ca^<2+>]_i. We conducted whole-cell current recording combined with fluorescence measurement of intracellular Ca^<2+> concentration ([Ca^<2+>]_i) in order to investigate the mechanism underlying the C1-sensitive Ca^<2+> entry in endothelial cells. Membrane currents from single endothelial cells were measured using nystatin-perforated patch clamp technique. The histamine-induced Ca^<2+> entry is inhibited reversibly either by decreasing the extracellular concentration of Cl^- or by Cl^- channel blockers. Histamine activated an outward current, followed by a sustained inward current at -50 mV.The reversal potential was more negative than -60 mV for the initial outward current and approximately -30 mV for the sustained inward current with the normal Tyrode solution and the internal solution containing 30 epsilonlm Cl^-. The fluid stream applied through a micro tube also induced an inc … More rease in [Ca^<2+>]_i, which was dependent on both the flow rate and the extracellular Ca^<2+> concentration. The fluid stream-induced increase in [Ca^<2+>]_i was accompanied by the activation of an inward current at -53 mV.The reversal potential of the fluid stream-induced current shifted to positive potentials by reducing the external Cl^- concentration. Results from the effects of histamine and a fluid-stream we concluded that not only the histamine-induced inward current but also the fluid stream-induced current is carried mainly by Cl^-. And Cl^- current plays a crucial role in modulating the Ca^<2+>_i influx by altering the membrane potential of endothelial cells.
It is well known that sulfonylureas, which block ATP-sensitive K^<2+> channels (KATP), inhibit cystic fibrosis transmembrane regulator (CFTR) Cl^- channels, volume-sensitive Cl^- channels and Ca^<2+> activated Cl^- channels in epithelial and cardiac cells. Glibenclamide (10-500 muM) has little effect on the initial transient increase in [Ca^<2+>]_i induced by histamine but inhibited the following sustained increase in [Ca^<2+>]_i in a concentration-dependent manner. The IC50 value was 151.8 muM and the Hill's coefficient was 1.9. The sustained increase in [Ca^<2+>]_i induced by ATP (100 muM) and cyclopiazonic acid (10 muM) were also suppressed by glibenclamide, Under the patch-clampcondition, however, glibenclamide suppressed the histamine-induced Cl^- current with an IC50 value of 12 muM.Histamine-induced Cl^- current was almost completely abolished by 100 muM glibenclaminde but the sustainedincrease in [Ca^<2+>]_i was only partially suppressed. Thus, the glibenclamide-induced inhibition of the Ca^<2+> entry was not due to possible changes of membrane potential by suppression of Cl^- current, but due to direct effects on the Ca^<2+> influx pathway. The present results indicate that glibenclamide disturbs the Ca^<2+> influx pathway independent of the inhibition of Cl^- current, However, the activation of Cl^- channels might be functionally coupled with Ca^<2+> influx pathway in vascular endothelial cells. Less

Report

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

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Nakao, M., Ono, K., Fujisawa, S.& Iijima, T.: "Mechanostress-induced Ca2+ entry and C1- current in cultured human aortic endothelial cells." Am.J.Physiol.276. C238-C249 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ono, K., Nakao, M & Iijima, T.: "Chloride sensitive nature of histamine-induced Ca2+ entry in cultured human aortic endothelial cell." J.Physiol.511. 837-849 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ono, K., Nakao, M. & Iijima, T.: "C1- and voltage-dependent Ca2+ entry in cultured human aortic endothelial cell." Heart and Vessels. 12. 50-52 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 飯島 俊彦: "Store-operated Channel (SOC)とTransient Receptor Potential (TRP.)" 日本薬理学会誌. 112(5). 334 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 飯島 俊彦: "Ca^<2+>通過性チャネル" 現代医療, 特集「イオンチャネルと疾患」. 31(印刷中). 4 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nakao, M., Ono, K., Fujisawa, S.& Iijima, T.: "Mechanostress-induced Ca^<2+> entry and Cl^- current in cultured human aortic endothelial cells." Am.J.Physiol. 276. C238-C249 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ono, K., Nakao, M.& Iijima, T.: "Chloride sensitive nature of histamine-induced Ca^<2+> entry in cultured human aortic endothelial cell." J.Physiol.511. 837-849 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ono, K., Nakao, M.& Iijima, T.: "Cl^- and voltage-dependent Ca^<2+> entry in cultured human aortic endothelial cell." Heart and Vessels. 12. 50-52 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Iijima, T.: "Store-operated Channel (SOC) and Transient Receptor Potential (TRP)." Folia Pharmacol.Japon. 112. 334 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Iijima, T.: "Ca^<2+> permeable channel. (in press)." Gendai-iryo. 31 (4). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nakao,M.,One,K.,Fujisawa,S.& Iijima,Y.: "Mechanostress-induced Ca2+ entry and Cl- current in cultured human aortic endothelial cells." Am.J.Physiol.276. C238-C249 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ono,K.,Nakao,N.& Iijima,T.: "Chloride sensitive nature of histamine-induced Ca2+ entry in cultured human aortic endothelial cell." J.Physiol. 511. 837-849 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ono,K.,Nakao,M.& Iijima,T.: "Cl- and voltage-dependent Ca2+ entry in cultured human aortic endothelial cell." Heart and Vessels. 12. 50-52 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] 飯島 俊彦: "「Ca2+通過チャネル」" 現代医療, 特集「イオンチャネルと疾患」. 31. (4) (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ono, K., Nakao,M., and Iijima,T.: "Chloride-and voltage-dependent Ca^<2+> transient in cultured human aortic endothelial cell." Heart Vessels. Suppl. 12. 50-52 (1997)

    • Related Report
      1997 Annual Research Report

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

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