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2000 Fiscal Year Final Research Report Summary

Immunocytochemical Study on the Redistribution of Signal Molecules in Human Platelet Adherent to Extracellular Matrices

Research Project

Project/Area Number 11671021
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Hematology
Research InstitutionTokyo metropolitan Organization for Medical Research

Principal Investigator

SUZUKI Hidenori  Department of Cardiovascular Research, The Tokyo Metropolitan Institute of Medical Science, Researcher, 東京都臨床医学総合研究所, 研究員 (30158977)

Co-Investigator(Kenkyū-buntansha) AOKI Kazumasa  Department of Cardiovascular Research, The Tokyo Metropolitan Institute of Medical Science, Researcher, 東京都臨床医学総合研究所, 研究員 (10184029)
Project Period (FY) 1999 – 2000
Keywordsplatelets / adhesion / focal adhesion / immunoelectron microscopy / tyrosine phosphorylation / Src / Syk / FAK / P13-kinase
Research Abstract

Using immunofluorescence microscopy, we have examined the cytoskeletal patterns in human platelets adherent to collagen. Vinculin was detected as aligned patches, and located along actin filaments. Although talin was not detected at the ends of actin filaments, it was concentrated at the periphery of the platelets where the platelets were in close contact with adjacent platelets. pp125^<FAK> was detected as small spots, all of which, as a whole, formed a ring at the central areas of adherent platelets. These results suggest that the adherent platelets fail to form typical focal contacts.
Focal adhesion kinase (FAK) is phosphorylated on tyrosine during agonist-induced platelet aggregation or spreading on matrices. In this study, using a immunocytochemical method, we focused on (re)distribution of FAK during platelet activation and aggregation induced by thrombin. We found that FAK translocates to submembrane zone just beneath plasma membrane from cytoplasm prior to both aggregation and tyrosine phosphorylation of the kinase.
Next, we have examined the role of phosphoinositide 3-kinase (P13-K) in platelet adhesion and aggregation using wortmannin and LY294002, two unrelated inhibitors of P13-K.Pretreatment of the platelets with the inhibitors prevented platelet spreading on collagen and aggregation induced by thrombin receptor activator peptide. These results suggest that P13-K is necessary for the activation of αllbβ3 integrin and maintenance of the integrin in its active state.

  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Hidenori Suzuki: "Down-regulation and redistribution of GPV/GPVf2, a subunit of von Willebrand factor receptor (GPIb/IX/V complex), on the surface membrane of thrombin-stimulated human platelets."Br J Haematol. 104. 55-63 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hidenori Suzuki: "The behavior of calpain-generated N- and C-terminal fragments of talin in integrin-mediated signaling pathways."Arch Biochem Biophys. 371. 133-141 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hidenori Suzuki: "Translocation of sLe^xon the azurophilie granule membrane to the plasma membrane in activated human neutrophils."J Electron Microsc. 49. 359-370 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hidenori Suzuki: "血小板粘着・凝集機構におけるPI3-キナーゼの役割の検討-電子顕微鏡的研究-"日本血栓止血学会誌. 11. 255-264 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hidenori Suzuki: "Junctional adhesion molecule (JAM) is phosphorylated by protein kinase C upon platelet activation."Biochem Biophys Res Commun. 276. 873-878 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawano, H., Suzuki, H., Tanoue, K., Kimura, A., Fujimura, K.: "Down-regulation and redistribution of GPV/GPVf_2, a subunit of von Willebrand factor receptor (GP1b/1X/V complex), on the surface membrane of thrombin-stimulated human platelets."Br. J.Hamatol.. 104. 55-63 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hayashi, M., Suzuki, H., Kawashima, S., Saido, T.C., Inomata, M.: "The behavior of calpaingenerated N- and C-terminal fragments of talin in integrin-mediated signaling pathways."Arch. Biochem. Biophys.. 371. 133-141 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suzuki, H., Yokomizo, S., Wakamoto, S., Watanabe, K., Akamatsu, N., Tanoue, K.: "Translocation of sLe^x on the azurophilic granule membrane to the plasma membrane in activated human neutrophils."J.Electron Microsc.. 49. 359-370 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ozaki, H., Ishii, K., Arai, H., Horiuchi, H., Kawamoto, T., Suzuki, H., Kita, T.: "Junctional adhesion molecule (JAM) is phosphorylated by protein kinase C upon platelet activation."Biochem. Biophys. Res. Commun.. 276. 873-878 (2000)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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