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The functional development of excitation-contraction coupling during cardiomyogenesis in mouse embryonic stem cells

Research Project

Project/Area Number 13670037
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General physiology
Research InstitutionTokyo Medical & Dental University

Principal Investigator

KAWANO Seiko  Department of Cardiovascular Diseases, Medical Research Institute, Associate Professor, 難治疾患研究所, 助教授 (00177718)

Co-Investigator(Kenkyū-buntansha) YAMAGATA Kazuo  Division of Life Science, Graduated School of Integrated Science and Art, University of East Asia, 工学部, 助教授 (10299323)
ICHINOSE Shizuko  Instrumental Analysis Research Center, Tokyo Medical and Dental University, Assistant Professor, 機器分析センター, 助手 (60014156)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2002: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2001: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsES cell / cardiomyocyte / excitation-contraction coupling / IP3 receptor / Ryanodine receptor / Ca^<2+> signaling pathway / Na^+ Ca^<2+> exchanger / Na^+・K^+ pump / 胚性幹細胞 / Caシグナル伝達 / plasma membrane Ca pump / Na-Ca exchanger / Na-K pump / 幹細胞 / カルシュウム / カルシュウムチャネル / 容量性カルシュウム流入 / 分化 / 心筋
Research Abstract

Mouse embryonic stem (ES) cells differentiate to all cell lineages including cardiomyocytes. In this study, we investigated the functional development of excitation-contraction coupling during cardiomyogenesis in mouse embryonic stem cells. During the differentiation to cardiomyocytes from ES cells (Cell line D3), the Ca^<2+> signaling pathway in each stage was examined using Ca^<2+> or Na^+ imaging, patch clamp techniques and RT-PCR. Our results showed that in undifferentiated ES cells Ca^<2+> was released from ER by the stimulation of G-protein coupled receptors. Using RT-PCR, mRNA for IP3 type I, II and III were detected, confirming IP3-induced Ca^<2+> release (IICR). On the other hand, function of ryanodine receptors or voltage-operated Ca^<2+> channels (VOCC) could not be detected, indicating no function of Ca^<2+>-induced Ca^<2+> release (CICR). For the Ca^<2+> entry pathway, store-operated Ca^<2+> entry (ISOC) was observed. From these results we conclude that Ca^<2+> entry through plasma membrane is mainly mediated by ISOC with little contribution of VOCCs. In this study, we also found that Na^+-Ca^<2+> exchangers and plasma membrane pumps functioned as Ca^<2+> extrusion system. The function of Na^+-K^+ pump examined using ouabain sensitivity could be detected in ES cells. During the differentiation to cardiomyocytes, IICR function was observed in all stages. In the 7-day differentiated cells, CICR could be observed. Both T-type and L-type Ca^<2+> channel were recorded by patch clamp experiments. Ouabain-sensitive Na^+-K^+ pump functions were increased with the differentiation. In whole cell patch clamp, Na^+-K^+ pump currents could be recorded in cells from at 8-day-differentiated stage but not earlier stages. Thus, we clarified the Ca^<2+> signaling pathways in undifferentiated mouse ES cells for the first time in this study. However, the further studies are needed to explore the mechanisms how these signaling pathways differentiate or develop.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] Seiko Kawano et al.: "Ca^<2+> Oscillations regulated by Na^+ -Ca^<2+> Exchanger and Plasma Membrane Ca^<2+> Pump Induce Fluctuations of Membrane Currents and Potentials in Human Mesenchymal Stem Cells"Cell Calcium. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tagami M., Ichinose S., Yamagata K., Fujino H., Shoji S., Hiraoka M., Kawano S.: "Genetic and ultrastructural demonstration of strong reversibility in human mesenchymal stem cell"Cell and Tissue Research. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Seiko Kawano et al.: "Characterization of Ca^<2+> signaling pathway in human mesenchymal stem cells"Cell Calcium. 32. 165-174 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hirayama Y., Kuruma A., Hiraoka M., Kawano S: "Ca^<2+>-activated Cl^-current is enhanced by acidosis and contributes to the shortening of action potential duration in rabbit ventricular myocytes"Japanese Journal of Physiology. 52. 293-300 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hirayama Y., Kuruma A., Hiraoka M., Kawano S.: "Beat dependent alteration of Ca-activated Cl current during rapid stimulation in rabbit ventricular myocytes"Japanese Heart Journal. 42. 207-219 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 川野誠子: "心筋クロライドチャネルと不整脈"臨床と研究. 78. 149-219 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Masayasu Hiraoka, Yuji Hirano, Seiko Kawano, Tetsushi Furukawa: "Heart Physiology and Pathophysiology"Regulation of cardiac Ion channels by phosphrylation, Ca, Cytoskeleon, and Stretch. 389-404 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Seiko Kawano et al: "Ca^<2+> Oscillations regulated by Na^+-Ca^<2+> Exchanger and Plasma Membrane Ca^<2+> Pump Induce Fluctuations of Membrane Currents and Potentials in Human Mesenchymal Stem Cells"Cell Calcium. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tagami M., Ichinose S., Yamagata K., Fuiino H., Shoii S., Hiraoka M., and Kawano S.: "Genetic and ultrastructural demonstration of strong reversibility in human mesenchymal stem cell."Cell and Tissue Research. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Seiko Kawano et al: "Characterization of Ca^<2+> signaling pathway in human mesenchymal stem cells"Cell Calcium. 32. 165-174 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hirayama Y., Kuruma A., Hiraoka M., and Kawano S.: "Ca^<2+>-activated CT current is enhanced by acidosis and contributes to the shortening of action potential duration in rabbit ventricular myocytes"Japanese Journal of Physiology. 52. 293-300 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hirayama Y., Kuruma A., Hiraoka M., and Kawano S.: "Beat dependent alteration of Ca-activated Cl current during rapid stimulation in rabbit ventricular myocytes"Japanese Heart Journal. 42. 207-219 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Masayasu Hiiaoka, Yuji Hirano, and Seiko Kawano, Tetsushi Furukawa: "Regulation of cardiac Ion channels by phosphrylation, Ca, Cytoskeleon, and Stretch"Heart Physiology and Pathophysiology. 389-404 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Seiko Kawano et al.: "Ca^<2+> Oscillations regulated by Na^+-Ca^<2+> Exchanger and Plasma Membrane Ca^<2+> Pump Induce Fluctuations of Membrane Currents and Potentials in Human Mesenchymal Stem Cells"Cell Calcium. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Seiko Kawano et al.: "Characterization of Ca2+ signaling pathways in human mesenchymal stem cells"Cell Calcium. 32. 165-174 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hirayama Y., Kuruma A., Hiraoka M., Kawano S.: "Ca-activated Cl current is enhanced by acidosis and contributes to the shortening of action potential duration in rabbit ventricular myocytes"Japanese Journal of Physiology. 52. 293-300 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tagami M., Ichinose S., Yamagata K., Fujino, H., Shoji S., Hiraoka M., Kawano S: "Genetic and ultrastructural demonstration of strong reversibility in human mesenchymal stem cell"Cell and Tissue Research. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hirayama Y., Kuruma A., Hiraoka M., Kawano S.: "Beat dependent alteration of Ca-activated Cl current during rapid stimulation in rabbit ventricular myocytes"Japanese Heart Journal. 42. 207-219 (2001)

    • Related Report
      2002 Annual Research Report
  • [Publications] 川野誠子: "心筋のクロライドチャネルと不整脈"臨床と研究. 78. 149-219 (2001)

    • Related Report
      2002 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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