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Molecular Anatomy of Gap Junction Expression Regulation Effect in Conditional Cx Knockout Mice

Research Project

Project/Area Number 13470004
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General anatomy (including Histology/Embryology)
Research InstitutionKyushu University

Principal Investigator

SHIBATA Yosaburo  Kyushu University, Faculty of Medical Sciences, Professor, 大学院・医学研究院, 教授 (90037482)

Co-Investigator(Kenkyū-buntansha) NISHII Kiyomasa  Kyushu University, Faculty of Medical Sciences, Research Associate, 大学院・医学研究院, 助手 (20264020)
INAI Tetsuichiro  Kyushu University, Faculty of Medical Sciences, Lecture, 大学院・医学研究院, 講師 (00264044)
NAKAMURA Keiichiro  Kyushu University, Faculty of Medical Sciences, Associate Prof., 大学院・医学研究院, 助教授 (20172398)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥14,200,000 (Direct Cost: ¥14,200,000)
Fiscal Year 2002: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2001: ¥10,300,000 (Direct Cost: ¥10,300,000)
Keywordsgene targeting / heart / gap Junction / connexin / Cre / loxP / Cx 45 / embryonic cardiac myocyte / floxed : Cx mice / gene trageting / loxp
Research Abstract

1. We have created mice whose floxed-Cx45 locus can be removed conditionally. They are healthy when Cre is absent. However, cardiac α-actin-Cre-mediated Cx45-deletion resulted in early embryonic lethality. X-gal staining, a hallmark of Cre/loxP-mediated floxed-Cx45 deletion, was detected in the embryonic cardiac myocytes. In another experiment, embryonic cardiac myocytes were differentiated from Cx45-deficient ES cells in vitro. Their conduction velocity was slow, but contraction rhythm was rapid and irregular. Therefore we, for the first time, found that Cx45 is an essential gap junction protein required for the early embryonic cardiac myocytes.
2. On the other hand, smooth muscle α-actin-Cre-mediated Cx45-deletion yielded healthy mice, indicating that Cx45 is not essential in smooth muscle. However, Cx45 is so widely expressed in the smooth muscle cells that its detailed function on vascular smooth muscle remains to be clarified. We are examining how this vascular smooth muscle-specific Cx45 knockout affects the integrity and regulation of vascular tone.
3. The attempt to detect Cx43-Cx45 doubly deficient mice by a micro-drop culture system was not successful. At present, the fact seemingly indicates that a significant defect may impair early eggs when both connexins are lost. We are trying to create Cx43-KO/floxed-Cx45 mice, which will enable us to explore Cx43-Cx45 doubly deficient mice/cells in desired cell types at desired timing.
4. We have unraveled a detailed spatio-temporal expression profile of the Cx45-marker in the central nervous system. Strong expression in the thalamus was a novel finding. These data make ongoing collaboration very exciting (with Prof. David Paul at Harvard Medical School and Prof. Cecilia Lo at NIH, to create neuron/glia/neural crest-specific knockout of Cx45).

Report

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

    (27 results)

All Other

All Publications (27 results)

  • [Publications] H.Koga et al.: "Overexpression of Polycomb-group gene rae28 in cardiomyocytes does not complement abnormal cardiac morphogenesis in mice lacking rae28 but causes dilated cardiomyopathy"Lab. Invest.. 375-385 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M.Doiguchi et al.: "Complimentary DNA cloning and characterization of rat spergen-1, a spermatogenic cell-specific gene-1, containging a mitochondria-targeting signal"Biol.Reprod.. 66. 1462-1470 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] J.Kobayashi et al.: "Formation of tight junction strands by expression of claudin-1 mutants in their ZO-1 binding site in MDCK cells"Histchem. Cell Biol.. 67. 29-39 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M.Doiguchi et al.: "Spergen-1 might be an adhesive molecule associated with mitochondria in the middle piece of spermatozoa"Dev. Biol.. 252. 127-137 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] A.Kuraoka et al.: "Molecular cloning of two guinea pig liver gap junction proteins, connexin32 and connexin26"Fukuoka Acta Medica. 93. 178-188 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 中村 桂一郎 他: "ギャップ結合による細胞性ネットワーク -消化管運動ペースメーカー細胞を中心にして-"電子顕微鏡. 37. 188-193 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 柴田 洋三郎: "新女性医学体系 29巻 (器官の形成と分化)"中山書店. 397 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Koga H., Kaji Y., Nishii K., Shirai M., Tomotsune D., Osugi T., Sawada A., Kim JY., Hara J., Miwa T., Yamauchi-Takihara K., Shibata Y., Takihara Y.: "Overexpression of Polycomb-group gene rae28 in cardiomyocytes does not complement abnormal cardiac morphogenesis in mice lacking rae28 but causes dilated cardiomyopathy"Lab. Invest.. 82(4). 375-385 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M. Doiguchi, H. Yamashita, J. Ichinose, T. Mori, Y. Shibata, H. Iida: "Complementary DNA cloning and characterization of rat spergen-1, a spermatogenic cell-specific gene-1, containing a mitochondria-targeting signal"Biol. Reprod.. 66. 1462-1470 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] J. Kobayashi, T. Inai, and Y. Shibata: "Formation of tight junction strands by expression of claudin-1 mutants in their ZO-1 binding site in MDCK cells"Histochem. Cell Biol.. 117. 29-39 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Doiguchi M., Mori T., Toshimori K., Shibata Y., Iida H: "Spergen-1 might be an adhesive molecule associated with mitochondria in the middle piece of spermatozoa"Dev. Biol.. 252(1). 127-137 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] A. Kuraoka, T. Nakashima, M. Kawabuchi, Y. Shibata: "Molecular cloning of two guinea pig liver gap junction proteins, connexin32 and connexin26"Fukuoka acta medica. 93(9). 178-188 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Nakamura et.al.: "Interstitial cells of Cajal-gap junction connected celluilar networks as pacemaker system for gastrointestinal mortility"Denchi Kenbikyo. 37. 188-193 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y. Shibata: "Development of the organs"Shin Josei Igaku Taikei. 29. 1-397 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] H.Koga et al.: "Overexpression of Polycomb-group gene rae28 in cardiomyocytes does not complement abnormal cardiac morphogenesis in mice lacking rae28 but causes dilated cardiomyopathy"Lab. Invest.. 82・4. 375-385 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Doiguchi et al.: "Complementary DNA cloning and characterization of rat spergen-1, a spermatogenic cell-specfic gene-1, containing a mitochondria-targeting signal"Biol. Reprod.. 66・5. 1462-1470 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] J.Kobayashi et al.: "Formation of tight junction strands by expression of claudin-1 mutants in their ZO-1 binding site in MDCK cells"Histchem. Cell Biol.. 67. 29-39 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Doiguchi et al.: "Spergen-1 might be an adhesive molecule associated with mitochondria in the middle piece of spermatozoa"Dev. Biol.. 252・1. 127-137 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] A.Kuraoka et al.: "Molecular cloning of two guinea pig liver gap junction proteins, connexin32 and connexin26"Fukuoka Acta Medica. 93・9. 178-188 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 中村 桂一郎他: "ギャップ結合による細胞性ネットワーク-消化管運動ペースメーカー細胞を中心にして-"電子顕微鏡. 37・3. 188-193 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 柴田 洋三郎: "器官の形成と分化;新女性医学体系29"中山書店. 397 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] I.Yamanaka et al.: "Differential expression of major gap junction proteins, connexins26 and 32, in rat mammary glands during pregnancy and lactation"Histochem Cell Biol. 115. 277-284 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] H.Iida et al.: "Spermatid-specific expression of Iba 1, an ionized calcium binding adapter molecule-1, in rat testis"Biol Reprod. 64. 1138-1146 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Nishii et al.: "Regulation of the epithelial-mesenchymal transformation through gap junction channels in heart development"Trends Cardiovasc Med. 11. 213-218 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Y.Shibata et al.: "Diversity and molecular anatomy of gap junction"Med Electon Microsc. 34. 153-159 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] J.Kobayashi et al.: "Formation of tight junction strands by expression of claudin-1 mutants in their ZO-1 binding site in MDCK cells"Histochem Cell Biol.. 117. 29-39 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 柴田 洋三郎: "ギャップ結合の微細構造と分子生物学"電子顕微鏡. 36. 156-162 (2001)

    • Related Report
      2001 Annual Research Report

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

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