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Molecular and cell biolobical analysis of the smooth muscle cell differentiation

Research Project

Project/Area Number 07457029
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General medical chemistry
Research InstitutionOsaka University

Principal Investigator

SOBUE Kenji  Osaka Univ.Med.Sch.Prof., 医学部, 教授 (20112047)

Co-Investigator(Kenkyū-buntansha) TAKAMIYA Kougo  Osaka Univ.Med.Sch.Assistant.Prof., 医学部, 助手 (40283767)
HAYASHI Ken'ichiro  Osaka Univ.Med.Sch.Assistant.Prof., 医学部, 助手 (90238105)
乾 誠  大阪大学, 医学部, 助教授 (70223237)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1996: ¥1,200,000 (Direct Cost: ¥1,200,000)
Keywordssmooth muscle cell / phenotypic modulation / laminin / IGFI / caldesmon / alpha-tropomyosin / alpha1 integrin / alpha-smooth muscle actin / 転写制御 / 細胞接着 / 細胞外マトリクス / トロポミオシン / 遺伝子発現制御
Research Abstract

Primarily cultured smooth muscle cell (SMCs) rapidly dedifferentiate under normal culture conditions containing serum. We investigated to establish a culture system maintaining a differentiated phenotype of SMCs using several extracellular matrices and growth factors or cytokines. From these analyzes, we found that laminin has a potency to maintain a differentiated phenotype of SMCs under serum-free culture conditions. Furthermore, we obtained evidence that insulin-like growth factor I (IGFI), II (IGFII), or insulin possesses the remarkable activity to maintain a differentiated phenotype of SMCs for a long culture, and IGFI is a most potent factor for SMC differentiation, suggesting that signal transduction via laminin and IGFI receptors would be involved in SMC differentation. Pharmacological studies using several kinase inhibitors have revaled that tyrosine phosphorylation and PI3 kinase are involved in such signal transduction. We also found that the expression of IGFI-recepter is S … More MC phenotype-dependent. Therefore, the downregulation of IGFI-recepter might be one reason for dedifferentation of SMCs induced by serum growth factors. Using our SMC culture system, we characterized the SMC-specific gene expressions. In the caldesmon (CaD) gene, alternative selection of two 5'-splice sites within exon 3 determined h- or l-CaD expression. We found that hnRNPA1 is functionally involved in the selection of distal 5'-splice site. We found that expressional change of alpha-tropomyosin (alpha-TM) isoforms from alpha-TM-SM to alpha-TM-F1 and alpha-TM-F2 during dedifferentiation of SMCs and such conversion occurrs coordinately with the expressional change from h- to l-CaD,suggesting a common splicing mechanism for the phenotype-dependent expression of alpha-TM and CaD isoforms in SMCs. We also characterized transcriptional regulation of the CaD and the alpha1 integrin promoters in SMCs. These analyzes revealed that the CArG box within respective promoter regions are necessary for high level transcription of the both genes in differentiated SMCs, and the serum response factor (SRF) is a core factor for the CArG box binding. We demonstrated that the expression of alpha-SM actin in visceral SMCs is opposite to that in vascular SMCs ; alpha-SM actin is expressed in undifferentiated and dedifferentiated visceral SMCs, but not in differentiated visceral SMCs, and identified a novel cis-element in the promoter region of alpha-SM actin which acts as a negative regtulator. Less

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (45 results)

All Other

All Publications (45 results)

  • [Publications] Kira M.: "Caldesmon and low Mr isoform of tropomyosin are localized in neuronal growth cones." J. Neurosci. Res.40. 294-305 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yano H.: "Transcriptional regulation of the chicken caldesmon gene : activation of gizzard type caldesmon promoter requires a CArG box-like motif." J. Biol. Chem.270. 23661-23666 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yoshida K.: "Reperfusion of rat heart after brief ischemia induces proteolysis of calspectin (non-erythroid spectrin or fodrin) calpain." Circ. Res.77. 603-610 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ozawa K.: "Translocation of cortactin (p80/85) to the actin-based cytokeleton during thrombin receptor-mediated platelet activation." Exp. Cell Res.221. 197-204 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ilic D.: "Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice." Nature. 377. 539-544 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ozawa K.: "Phase specific association of heterotrimeric GTP binding proteins with the actin-based cytoskeleton during thrombin receptor mediated platelet activation." FEBS Lett.382. 159-163 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Akagi S.: "Localization of synapsin I in normal fibers and regenerating axonal sprouts of the rat sciatic nerve." Histochem. Cell Biol.105. 365-373 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kashiwada K.: "Coordinate expression of α-tropomyosin and caldesmon isoforms in association with phenotypic modulation of smooth muscle cells." J. Biol. Chem. (in press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ando Y.: "Enhanced glomerular expression of caldesmon in IgA nephropathy and its suppression by glucocorticoid therapy." J. Clin. Invest. (in press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Obata H.: "Smooth muscle cell phenotype-dependent transcriptional regulation of the α1 integrin gene." J. Biol. Chem. (in press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sobue K.: "Phenotype-dependent gene regulation of smooth muscle, cell-specific cytoskeletal proteins" Mol. Cell Biochem. (in press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hayashi K.: "IGF-I and laminin-generated signallings are essential for maintaining a differentiated phenotype of smooth muscle cells." J. Cell. Biol. (in press). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hayashi K.: "in Smooth Muscle Contraction. (Kohama K. and Saida K. ed. Japan Sci. Press, Tokyo." Expressional regulation of caldesmon isoforms in assoceation with phenotypic modulation of smooth muscle cells., 159 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kira M.: "Caldesmon and low Mr isoform of tropomyosin are localized in neuronal growth cones" J.Neurosci.Res.40. 294-305 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yano H.: "Transcriptional regulation of the chicken caldesmon gene : activation of gizzard type caldesmon promoter requires a CArG box-like motif" J.Biol.Chem.270. 23661-23666 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yoshida K.: "Reperfusion of rat heart after brief ischemia induces proteolysis of calspectin (non-erythroid spectrin or fodrin) by calpain" Circ.Res.77. 603-610 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ozawa K.: "Translocation of cortactin (p80/85) to the actin-based cytoskeleton during thrombin receptor-mediated platelet activation" Exp.Cell.Res.221. 197-204 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Illc D.: "Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice" Nature. 377. 539-544 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ozawa K.: "Phase specific association of heterotrimeric GTP binding proteins with the actin-based cytoskeleton during thrombin receptor mediated platelet activation" FEBS Lett.382. 159-163 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Akagi S.: "Localization of synapsin I in normal fibers and regenerating axonal sprouts of the rat sciatic nerve" Histochem.Cell Biol.105. 365-373 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kashiwada K.: "Coordinate expression of alpha-tropomyosin and caldesmon isoforms in association with phenotypic modulation of smooth muscle cells" J.Biol.Chem.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ando Y.: "Enhanced glomerular expression of caldesmon in IgA nephropathy and its suppression by glucocorticoid therapy" J.Clin.Invest.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Obata H.: "Smooth muscle cell phenotype-dependent transcriptional regulation of the alpha1 integrin gene" J.Biol.Chem.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sobue K.: "Phenotype-dependent gene regulation of smooth muscle, cell-specific cytoskeletal proteins" Mol.Cell.Biochem.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hayashi K.: "IGF-I and laminin-generated signallings are essential for maintaining a differentiated phenotype of smooth muscle cells" J.Cell.Biol.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hayashi K.: Smooth Muscle Contraction. (Kohama K.and Saida K.ed.), "Expressional regulation of caldesmon isoforms in assoceation with phenotypic modulation of smooth muscle cells". Japan Sci.Soc.Press, Tokyo., 131-141 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kira M.: "Caldesmon and low Mr isoform of tropomyosin are localized in neuronal growth cones." J.Neurosci.Res.40. 294-305 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Yano H.: "Transcriptional regulation of the chicken caldesmon gene : activation of gizzard type caldesmon promoter requires a CArG box-like motif." J.Biol.Chem.270. 23661-23666 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Yoshida K.: "Reperfusion of rat heart after brief ischemia induces proteolysis of calspectin (non-erythroid spectrin or fodrin) by calpain." Circ.Res.77. 603-610 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ozawa K.: "Translocation of cortactin (p80/85) to the actin-based cytoskeleton during thrombin receptor-mediated platelet activation." Exp.Cell.Res.221. 197-204 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ilic D.: "Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice." Nature. 377. 539-544 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ozawa K.: "Phase specific association of heterotrimeric GTP binding proteins with the actin-based cytoskeleton during thrombin receptor mediated platelet activation." FEBSL Lett.382. 159-163 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Akagi S.: "Localization of synapsin I in normal fibers and regenerating axonal sprouts of the rat sciatic nerve." Histochem.Cell Biol.105. 365-373 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kashiwada K.: "Coordinate expression of α-tropomyosin and caldesmon isoforms in association with phenotypic modulation of smooth muscle cells." J.Biol.Chem.(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ando Y.: "Enhanced glomerular expression of caldesmon in IgA nephropathy and its suppression by glucocorticoid therapy." J.Clin.Invest. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Obata H.: "Smooth muscle cell phenotype-dependent transcriptional regulation of the α1 integrin gene." J.Biol.Chem.(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Sobue K.: "Phenotype-dependent gene regulation of smooth muscle,cell-specific cytoskeletal proteins" Mol.Cell.Biochem.(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hayashi K.: "IGF-I and laminin-generated signallings are essential for maintaining a differentiated phenotype of smooth muscle cells." J.Cell.Biol.(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hayashi K.: "in Smooth Muscle Contraction. (Kohama K.and Saida K.ed.) Japan Sci.Soc.Press,Tokyo." Expressional regulation of caldesmon isoforms in assoceation with phenotypic modulation of smooth muscle cells., 159 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ilic D: "Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-dedicient mice." Nature. 377. 539-544 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yano H.: "Transcriptional regulation of chicken caldesmon gene : activation of gizzard type caldesmon promoter a CArG box-like motif." J.Biol.Chem.270. 23661-23666 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Ozawa K.: "Translocation of cortactin(p80/85) to the actin-based cytoskeleton during thrombin receptor-mediated platelet activation." Exp.Cell Res.221. 197-204 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yoshida K.: "Reperfusion of rat heart after brief ischemia induces proteolysis of calspectin (Non-erythroid spectrin or fodrin) by calpain." Circulation Res.77. 131-141 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kira M.: "Caldesmon and low Mr isoform of tropomyosin are localized in neuronal growth cones." J.Neurosci.Res.40. 294-305 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hayashi,K.: "Smooth Muscle Contraction." Expressional regulation of caldesmon isoforms in assoceation with phenotypic modulation of smooth muscle cells., 159 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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