Electron microscopic analyses on platelet membrane glycoproteins using monoclonal antibodies
Project/Area Number |
61570597
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Research Category |
Grant-in-Aid for General Scientific Research (C)
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Allocation Type | Single-year Grants |
Research Field |
Hematology
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Research Institution | The Tokyo Metropolitan Institute of Medical Science |
Principal Investigator |
YAMAZAKI Hiroh Vice Director The Tokyo Metropolitan Institute of Medical Science, 副所長 (50013826)
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Co-Investigator(Kenkyū-buntansha) |
山本 正雅雄 (山本 正雅) 東京都臨床医学総合研究所, 循環器病研究部, 研究員 (50150884)
TANOUE Kenjiro Dept.Director The Tokyo Metropolitan Institute of Medical Science Department of, 循環器病研究部, 研究部長 (30014137)
YAMAMOTO Naomasa Staff The Tokyo Metropolitan Institute of Medical Science Department of Cardiova
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Project Period (FY) |
1986 – 1987
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Project Status |
Completed (Fiscal Year 1987)
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Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
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Keywords | Platelet membrane glycoproteins / Immunocytochemistry / Platelet membrane recoptors / thrombin aggregation / ADP aggregation / fibrinogen / mooclonal antibody / トロンビン凝集 / 血小板無力症 / 受容体刺激 |
Research Abstract |
Processes of platelet adhesion and aggregation are mediated by specific membrane glycoproteins (GP) on the platelet surface.There are two kinds of binding sites; for von Willebrand factor and thrombin,on GPIb and fibriongen binding site on GPIIb/IIIa complex.We have examined distributions of GPs during platelet activation using immunocytochemical methods.TM60 which was produced by us using a hybridoma technique is a monoclonal antibody against GPIb. When glutaraldehyde-fixed platelets were incubated with TM60,a uniform distribution of ferritin particles which represent the localization of GPIb was observed on the surface membrane of platelets. The particles were distributed at intervals of about 100 nm.The numbers of ferritin particles on the surface of one side were 2070-4150(2940<plus-minus>790;mean<plus-minus>SD, n = 10) under the scanning electron microscope. The distribution was disarranged and cluster-like structures of particles were observed after the activation of platelets suggestion the role of GPIb as a receptor. Association of fibrinogen in ADP-and thrombin-induced platelet aggregation was observed using a goat antihuman fibrinogen antibody which reacted with a gold-labeled anti-goat IgG under the electron microscope.In resting platelets, gold particles were appearent in only aplha-granules and not on the platelet surface.When ADP was added.gold particles were detected on the platelet surface in the presence of added fibrinogen. ADP caused aggregaion of normal platelets and gold particles were evident between the adherent platelets and on the platelet surface.When the platelets deaggregated, gold was no longer present on the surface.In thrombin-induced aggregation, however,there are several aggregated masses without fibringen binding on the platelet surface.Thus fibrinogen may not be solely responsible to forming the links between thrombin-stimulated plateleffe
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Report
(2 results)
Research Products
(22 results)
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[Publications] Yamamoto,K,Yamamoto,N,Kitagawa,H,Tanoue,K,Kosaki,G,Yamazaki,H: "Localization of a thrombin-binding site on human platelet membrane glycoprotein Ib determined by a nomoclonal antibody" Thrombosis and Haemostasis. 55. 162-167 (1986)
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Related Report
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[Publications] Suzuki,H,Kinlough-Rathbone,RL,Packham,MA,Tanoue,K,Tamazaki,H, Mustard,JF: "Immunocytochemical localization of fibrinogen on washed human platelets. Lack of requirement for fibringen during ADP-induced responses,and enhanced fibringen binding in a medium with low calcium" Blood.
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「研究成果報告書概要(欧文)」より
Related Report
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[Publications] Suzuki,H,Kinlough-Rathbone,RL,Packham,MA,Tanoue,K,Yamazaki,H, Justard,JF: "Immunocytochemical localization of fibrinogen during thrombin-induced aggregation of washed human platelets" Blood.
Description
「研究成果報告書概要(欧文)」より
Related Report
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