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Disruption of focal adhesion during the recovery from in vitro ischemia

Research Project

Project/Area Number 06836021
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 血管生物学
Research InstitutionNATIONAL CARDIOVASCULAR CENTER RESEARCH INSTITUTE

Principal Investigator

MASUDA Michitaka  National Cardiovascular Center Research Institute, Department of Structural Analysis, Section Chief, 循環器形態部, 室長 (00190364)

Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsEndothelial cell / Ischemia-reperfusion / Cell-substrate adhesion / Focal adhesion / Tyrosine phosphorylation / PECAM-1
Research Abstract

We studied dynamic behavior of cell-substrate contacts, especially focal contacts (FCs), of vascular endothelial cells (ECs) cultured on a glass surface using an interference reflection microscope. ECs were cultured in our flow culture chamber and by completely stopping culture medium flow, we created an in vitro ischemic condition. The FC was a highly stable structure, but, when ECs in a confluent monolayr were kept under the in vitro ischemic condition for 2 hr and then exposed to flow, they transiently lost a significant number of their FCs. ECs detached from the substrate if the flow was fast enough to generate fluid shear stress of - 10 dyn/cm^2. To identify the stress (es) inducing the disruption of FCs, we exposed static EC cultures to conditions which were expected to occur in our in vitro ischemia-reperfusion experiment. ATP depletion, ionophore-induced Ca^<2+> influx, or oxidative stress (H_2O_2, t-butyl hydroperoxide, 15-HPETE) did not cause the loss of FCs. When ECs were pretreated for 40 min with a low pH medium (pH 6.3) containing 20 mM lactate and recovered in a normal pH medium without lactate, they transiently lost their FCs in the similar manner observed in the ischemia-reperfusion experiment. Concomitant with the FC loss, tyrosine phosphorylation of proteins around 120 kDa was augmented. We identified two of these proteins : one is pp125^<FAK> and the other is PECAM-1. Auti-phosphotyrosine staining intensity of the FC increased at the time of the FC loss. Treatment of ECs with tyrosine kinase inhibitors protected ECs from losing their FCs. These results suggest a regulatory role for protein tyrosine phosphorylation in the stability of FCs.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] 増田道隆: "血管内皮細胞の流れに対する応答とその機構" 蛋白質 核酸 酵素. 41. 34-47 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] N. Harada: "Fluid flow and osmotics induced tyrosine phosphorylation of an endothelial cell 128 KDa surface glycoprotein." Biochem. Biophysc. Res. Commun.214. 69-74 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K. Katoh: "Focal adhesion proteins associated with apical stress fiber of human fibroblasts." Cell Motil. Cytoskeleton. 31. 177-195 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 増田道隆: "培養血管内皮細胞の流れ応答性" 循環器病研究の進歩. 31. 84-94 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 増田道隆: "血管内皮細胞は血液の流れにどのように応答するか" 化学と生物. 32. 386-390 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 藤原敬己: "細胞骨格と接養因子" 腎と透析. 37. 451-458 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 増田道隆(分担執筆): "新しい光学顕微鏡(第2巻)レーザー顕微鏡の医学・生物学への応用" 学際企画, 199 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M. Masuda(分担執筆): "Progress in Microcirculation Research" Elsevier Science Ltd., 521 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Masuda, K.Katoh, K.Fujiwara: "Mechanisms for flowinduced responses of vascular endothelial cells.(in Japanese)" Protein, Nucleic Acid and Enzyme. 41. 34-47 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] N.Harada, M.Masuda, K.Fujiwara: "Fluid flow and osmotic stress induced tyrosine phosphorylation of an endothelial cell 128 kDa surface glycoprotein." Biochem.Biophysc.Res.Commun.214. 69-74 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Katoh, M.Masuda, Y.Kano, Y.Jinguji, K.Fujiwara: "Focal adhesion proteins associated with apical stress fibers of human fibroblasts." Cell Motil. Cytoskeleton. 31. 177-195 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Masuda, K.Fujiwara: "Flow-induced responses of vascular endothelial cells in vitro. (in Japanese)" Recent Advances in Cardiovascular Research. 31. 84-94 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Masuda, K.Fujiwara: "How vascular endothelial cells respond to blood flow." KASEAA. 32. 386-390 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Fujiwara, Y,Jinguji, M.Masuda: "The cytoskeleton and cell adhesion molecules." Kideny and Dialysis. 37. 451-458 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Masuda, K.Fujiwara: Observation of cell-substrate adhesion by interference reflection imaging.(in Japanese) in "New Optical Microscopy (Vol.2), Applications of confocal microscope for medical and biological researches" (S.Fujita, ed.). Gakusai Kikaku, Tokyo, 165-172 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Masuda, K.Fujiwara: Rapid Motile responses of single endothelial cells exposed to shear stress. in "Progress in Microcirculation Research" (H.Niimi et al. eds.). Elsevier Science Inc.Oxford, 203-207 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 増田 道隆: "血管内皮細胞の流れに対する応答とその機構" 蛋白質 核酸 酵素. 41. 34-47 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] N. Harada: "Fluid flow and osmotic stress induced tyrosine phosphorylation of on EC 128kDa serface gycoprotein." Biochem. Biophysc. Res. Commun.214. 69-74 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K. Katoh: "Focal adhesion proteins associated with apical stress fibers of human fibroblasts." Cell Motil. Cytosbeleton. 31. 177-195 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 増田 道隆: "培養血管内皮細胞の流れ応答性" 循環器病研究の進歩. 31. 84-94 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 増田 道隆: "血管内皮細胞は血液の流れにどのように応答するか" 化学と生物. 32. 386-390 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 藤原 敬己: "細胞骨格と接着因子" 腎と透析. 37. 451-458 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 増田道隆(分担執筆): "新しい光学顕微鏡(第2巻)レーザー顕微鏡の医学・生物学への応用" 学際企画, 199 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] M. Masuda(分担執筆): "Progress in Microcirculation Research" Elsevier Science Ltd., 521 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 増田 道隆: "血管内皮細胞は血液の流れにどのように応答するか" 化学と生物. 32. 386-390 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 藤原 敬己: "細胞基質間接着の構造" 生体の科学. 49. 349-356 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] 藤原 敬己: "血流と内皮細胞の細胞骨格" 血管と内皮. 4. 56-63 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Katoh,K.: "Focal adhesion proteins associated with apical stress fibers of human fibroblasts." Cell Motility and Cytoskeleton. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Masuda,M(分担執筆): "Progress in Microcirculation Research(H.Niimi et al.eds)" Elsevier Science Ltd., 521 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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