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Real time imaging analysis of platelets aggregation and thrombus formation under shear stress

Research Project

Project/Area Number 18590204
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General physiology
Research InstitutionHamamatsu University School of Medicine

Principal Investigator

URANO Tetsumei  Hamamatsu University School of Medicine, 医学部, Professor (50193967)

Co-Investigator(Kenkyū-buntansha) MOGAMI Hideo  Hamamatsu Univ. Sch of Med, 医学部, Associate Professor (90311604)
IHARA Hayato  Hamamatsu Univ. Sch of Med, 医学部, Assistant Professor (00223298)
SUZUKI Yuko  Hamamatsu Univ. Sch of Med, 医学部, Assistant Professor (20345812)
Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,810,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥210,000)
Fiscal Year 2007: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2006: ¥2,900,000 (Direct Cost: ¥2,900,000)
Keywordsthrombus formation / intra vital confocal microscopy / vo Willebrand Factor / ADAMTS13 / Thrombotic Thrombocytopenic Purpura / platelet / vascular endothelinl cell / von Willebrand因子
Research Abstract

[Background] After secretion from Weibel-Palade body in vascular endothelial cells (VECs), von Willebrand factor (vWF) circulates as ultra-large multimer forms (ULM-vWF) and adheres to subendothelial collagen, to which platelets subsequently adheres to form platelet thrombi. Lacking of vWF leads to bleeding tendency (vWF disease) and excess amounts of ULM-vWF leads to thrombotic disease (thrombotic thrombocytopenic purpura). [Aim] Employing gene deficiency mouse of vWF cleaving enzyme (ADAMTS13) hybridized with green fluorescent protein (GFP) expressing transgenic mouse (KO mouse), we analyzed the dynamics of vWF exocytosis as well as platelet adhesion and its aggregation under shear stress by intra-vital confocal microscopy. [Results] (1) DDAVP, known to stimulate vWF exocytosis, increased ULM-vWF which was visualized as long strings using fluorescent labeled anti-vWF antibody. (2) The strings were longest at 5th minute, and longer in KO mice (5.28±4.28 vs 2.89±2.08 μm). (3) GFP expressing platelets adhered to being secreted ULM-vWF. (4) After topical infusion of 2.5% FeC12, longer (26.34±8.28 vs 10.73±3.11 μm) and more stable (2.35±1.23 vs 0.63±0.21 s) strings were visualized in KO mice. [Conclusion] (1) Platelets adhesion and aggregation were successfully monitored by intra-vital confocal microscopy. (2) ADAMTS13 appeared to cleave ULM-vWF under shear stress when it is being secreted. (3) The enhanced secretion itself, however, did not develop microthrombus even in KO mice. (4) Longer and more stable ULM-vWF adhered to injured VECs by ferric chloride treatment in KO mice. (5) Larger thrombus was formed in KO-mice at the initial phase after laser-irradiated injury of vascular wall, whereas there was no difference in the size of stabilized thrombus. (6) ADAMTS13 was shown to be responsible for the cleavage of ULM-vWF under shear stress, and lack of its activity appeared to be thrombogenic.

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (39 results)

All 2008 2007 2006 Other

All Journal Article (17 results) (of which Peer Reviewed: 7 results) Presentation (19 results) Book (3 results)

  • [Journal Article] ウロキナーゼ,ヘパリン併用被覆カテーテルの血栓形成阻害効果の検討2008

    • Author(s)
      三上 英智, 小林 隆夫, 浦野 哲盟
    • Journal Title

      日本血栓止血学会誌 19(2)

      Pages: 257-264

    • NAID

      10026116182

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] 侵襲時の線溶活性=病態増悪因子としてのplasminogen activator inhibitor type 1(PAI-1)とプロテアーゼインヒビター=2007

    • Author(s)
      浦野哲盟
    • Journal Title

      Surgery Frontier 14(1)

      Pages: 109-113

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] 血漿 tPA 及びPAI-1抗原量測定の意義 -新たな知見からの再考察-2007

    • Author(s)
      鈴木優子、浦野哲盟
    • Journal Title

      日本血栓止血学会雑誌、18(3), 247-254, 2007 18(3)

      Pages: 247-254

    • NAID

      10026114815

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] 抗炎症療法と血管病予防の関連を探る2007

    • Author(s)
      浦野哲盟
    • Journal Title

      Vascular Medicine 3(4)

      Pages: 333-338

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Role of tPA in the neural system2007

    • Author(s)
      Nagai, N., Urano, T
    • Journal Title

      In Recent Advances in Thrombosis and Hemostasis. Edited by K. Tanaka and E.W. Date. Springer,Tokyo 643

      Pages: 314-327

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] 侵襲時の線溶活性=病態増悪因子としてのplasminogen activator inhibito rtype1(PAI-1)とプロテアーゼインヒビター=2007

    • Author(s)
      浦野 哲盟
    • Journal Title

      Surgery Frontier 14(1)

      Pages: 109-113

    • Related Report
      2007 Annual Research Report
  • [Journal Article] 血漿tPA及びPAI-1抗原量測定の意義-新たな知見からの再考察-2007

    • Author(s)
      鈴木 優子、浦野 哲盟
    • Journal Title

      日本血栓止血学会雑誌、18(3), 247-254, 2007 18(3)

      Pages: 247-254

    • NAID

      10026114815

    • Related Report
      2007 Annual Research Report
  • [Journal Article] 抗炎症療法と血管病予防の関連を探る2007

    • Author(s)
      浦野 哲盟
    • Journal Title

      Vascular Medicine 3(4)

      Pages: 333-338

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Less pronounced enhancement of thrombin-dependent inactivation of plasminogen activator inhibitor type 1 by low molecular weight heparin compared with unfractionated heparin.2006

    • Author(s)
      Nakamura R, Umemura K, Hashimoto H, Urano T
    • Journal Title

      Thrombosis and Haemostasis 95

      Pages: 637-642

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary 2006 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Glucagon-Like Peptide 1 Activates Protein Kinase C Through a Ca^<2+>-Dependent Phospholipase C Activation in Insulin-Secreting Cells2006

    • Author(s)
      Suzuki Y, Urano T, Zhang H, Saito N, Kojima N, Mogami H
    • Journal Title

      Journal of Biological Chemistry 281・39

      Pages: 28499-28507

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary 2006 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 線溶療法2006

    • Author(s)
      浦野哲盟
    • Journal Title

      日本血栓止血学会誌 17・6

      Pages: 676-681

    • NAID

      10021259521

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary 2006 Annual Research Report
  • [Journal Article] Real-time analysis of platelet aggregation and procoagulant activity during thrombus formation in vivo.

    • Author(s)
      Hayashi T, Mogami H, Murakami Y, Nakamura T, Kanayama N, Konno K, Urano T.
    • Journal Title

      Pflugers Archiv-European Journal of Physiology (In press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Intraperitoneal Infusion of Recombinant Plasminogen Activator inhibitor Type 2 Induced Apoptosis in Implanted Human Colon Cancer and Inhibited its Growth and Liver Metastasis.

    • Author(s)
      Suwa D, Konno H, Tanaka T, Urano T.
    • Journal Title

      Anticancer Research (In press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Real-time analysis of platelet aggregation and procoagulant activity during thrombus formation in vivo

    • Author(s)
      Hayashi, T., Mogami, H., Murakami, Y., Nakamura, T., Kanayama, N., Konno, K., Urano, T
    • Journal Title

      Pflugers Archiv-European Journal of Physiology (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Intraperitoneal Infusion of Recombinant Plasminogen Activator inhibitor Type 2 Induced Apoptosis in Implanted Human Colon Cancer and Inhibited its Growth and Liver Metastasis

    • Author(s)
      Suwa, D., Konno, H., Tanaka, T., Urano, T
    • Journal Title

      Anti Cancer Research (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Real-time analysis of platelet aggregation and procoagulant activity during thrombus form ation in vivo

    • Author(s)
      Hayashi T, Mogami H, Murakami Y, Nakamura T, Kanayama N, Konno K, Urano T.
    • Journal Title

      Pflugers Archiv-European Journal of Physiology (In press)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Intraperitoneal Infusion of Recombinant Plasminogen Activator inhibitor Type 2 Induced Apoptosis in Implanted Human Colon Cancer and Inhibited its Growth and Liver Metastasis

    • Author(s)
      Suwa D, Konno H, Tanaka T, Urano T.
    • Journal Title

      Anticancer Research (In press)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] 血液及び血管内皮から見た血栓症リスク シンポジウム「深部静脈血栓症の病態生理とその予防」2008

    • Author(s)
      浦野哲盟、鈴木優子、井原勇人、最上秀夫
    • Organizer
      第85回日本生理学会
    • Place of Presentation
      東京
    • Year and Date
      2008-03-26
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 血液及び血管内皮から見た血栓症リスクシンポジウム「深部静脈血栓症の病態生理とその予防」2008

    • Author(s)
      浦野 哲盟、鈴木 優子、井原 勇人、最上 秀夫
    • Organizer
      第85回日本生理学会
    • Place of Presentation
      東京
    • Year and Date
      2008-03-26
    • Related Report
      2007 Annual Research Report
  • [Presentation] 血栓溶解薬の開発状況 シンポジウム「血栓症治療薬の現状と新薬の開発状況」2008

    • Author(s)
      浦野哲盟
    • Organizer
      第128回日本薬学会
    • Place of Presentation
      横浜
    • Year and Date
      2008-03-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 血栓溶解薬の開発状況シンポジウム「血栓症治療薬の現状と新薬の開発状況」2008

    • Author(s)
      浦野 哲盟
    • Organizer
      第128回日本薬学会
    • Place of Presentation
      横浜
    • Year and Date
      2008-03-25
    • Related Report
      2007 Annual Research Report
  • [Presentation] 接触因子によるPAI-1不活性化に伴う線溶活性増強機構2007

    • Author(s)
      田中晶、鈴木優子、金山尚裕、浦野哲盟
    • Organizer
      第30回日本血栓止血学会
    • Place of Presentation
      志摩市
    • Year and Date
      2007-11-17
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [Presentation] 生体内顕微鏡を用いた血栓形成時の血小板phosphatidylserine発現の時空間的調節機構の解析 シンポジウム「脂質ダイナミズムによる血栓止血反応の制御」2007

    • Author(s)
      林 忠毅, 最上秀夫, 村上裕介, 鈴木優子、今野弘之, 浦野哲盟
    • Organizer
      第30回日本血栓止血学会
    • Place of Presentation
      志摩市
    • Year and Date
      2007-11-15
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 生体内顕微鏡を用いた血栓形成時の血小板phosphatidylserine発現の時空間的調節機構の解析シンポジウム「脂質ダイナミズムによる血栓止血反応の制御」2007

    • Author(s)
      林 忠毅, 最上 秀夫, 村上 裕介, 鈴木 優子、今野 弘之, 浦野 哲盟
    • Organizer
      第30回日本血栓止血学会
    • Place of Presentation
      志摩市
    • Year and Date
      2007-11-15
    • Related Report
      2007 Annual Research Report
  • [Presentation] Modification by PAI-1 0f slow exocytotic dynamics of tissue plasminogen activator(tPA)and its activity on vascular endothelial cells(VECS)2007

    • Author(s)
      Urano T, Suzuki Y, Ihara H, Mogami H.
    • Organizer
      XXIth Congress of the International Society on Thrombosis and Haemostasis
    • Place of Presentation
      Geneva
    • Year and Date
      2007-07-09
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Modification by PAI-1 of slow exocvtotic dynamics of tissue plasminogen activator (tPA) and its activity on vascular endothelial cells (VECS)2007

    • Author(s)
      Urano, T., Suzuki, Y., Ihara, H., Mogami, H
    • Organizer
      XXIth Congress of the International Society on Thrombosis and Haemostasis
    • Place of Presentation
      Geneva
    • Year and Date
      2007-07-09
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Modification by PAI-1 of slow exocytotic dynamics of tissue plasminogen activator(tPA)and its activity on vascular endothelial cells(VECS)2007

    • Author(s)
      Urano T, Suzuki Y, Ihara H, Mogami H.
    • Organizer
      XXIth Congress of the International Society on Thrombosis and Haemostasis
    • Place of Presentation
      Geneva
    • Year and Date
      2007-07-09
    • Related Report
      2007 Annual Research Report
  • [Presentation] nique Secretory Dynamics of Tissue Plasminogen Activator(tPA)is Beneficial to Maintain Fibrinolytic Activity on Cell Surface2007

    • Author(s)
      Suzuki Y, Ihara H, Mogami H, Urano T
    • Organizer
      XIth International Workshop on Molecular & Cellular Biology of Plasminogen Activation
    • Place of Presentation
      Stockholm
    • Year and Date
      2007-06-17
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Unique Secretory Dynamics of Tissue Plasminogen Activator (tPA) is Beneficial to Maintain Fibrinolytic Activity on Cell Surface2007

    • Author(s)
      Suzuki, Y., Ihara, H., Mogami, H., Urano, T
    • Organizer
      XIth International Workshop on Molecular & Cellular Biology of Plasminogen Activation
    • Place of Presentation
      Stockholm
    • Year and Date
      2007-06-17
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] nique Secretors Dynamics of Tissue Plasminogen Activator(tPA)is Beneficial to Maintain Fibrinolytic Activity on Cell Surface2007

    • Author(s)
      Suzuki Y, Ihara H, Mogami H, Urano T
    • Organizer
      XIth International Workshop on Molecular & Cellular Biology of Plasminogen Activation
    • Place of Presentation
      Stockholm
    • Year and Date
      2007-06-17
    • Related Report
      2007 Annual Research Report
  • [Presentation] Plasminogen activator inhibitor 1 forms a stable non-acyl enzyme inhibitor complex with tissue plasminogen activator2006

    • Author(s)
      T. Yamashita, Q. Dai, H. Ihara, Y. Suzuki, H. Mogami, T. Koide, F. J. Castellino and T. Urano
    • Organizer
      18th Congress on Fibrinolysis & Proteolysis
    • Place of Presentation
      San Diego
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 開口放出過程の可視化により判明したtPA分子特有の分泌動態と酵素活性発現調節 シンポジウム「細胞線溶の新展開」2006

    • Author(s)
      鈴木優子、浦野哲盟
    • Organizer
      第29回日本血栓止血学会
    • Place of Presentation
      宇都宮
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ヘパリン、ウロキナーゼ併用被覆カテーテルの血栓形成阻害効果の検討2006

    • Author(s)
      三上英智, 小林隆夫, 浦野哲盟
    • Organizer
      第29回日本血栓止血学会
    • Place of Presentation
      宇都宮
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Kinetics of tissue plasminogen activator(tPA)exocytosis from endothelial cells and its modulation by PA inhibitor-1(PAI-1),2006

    • Author(s)
      Urano T, Suzuki Y, Ihara H, Mogami H
    • Organizer
      The 4th Asian-Pacific Congress on Thrombosis and Hemostasis
    • Place of Presentation
      Suzhou, China
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 血管内血小板凝集能と凝固能の in vivo real time imag ing2006

    • Author(s)
      林忠毅、最上秀夫、村上祐介、浦野哲盟
    • Organizer
      第14回日本血管生物医学会
    • Place of Presentation
      東京
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 全反射蛍光顕微鏡を用いた血管内皮細胞からの組織型プラスミノーゲンアクチベーター開口放出動態のリアルタイム解析2006

    • Author(s)
      鈴木優子、井原隼人、浦野哲盟
    • Organizer
      第14回日本血管生物医学会
    • Place of Presentation
      東京
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Book] Role of tPA in the neural system. In Recent Advances in Thrombosis and Hemostasis. Edited by K. Tanaka and E.W. Davie.2007

    • Author(s)
      Nagai, N. and Urano, T.
    • Publisher
      Springer, Tokyo
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Book] 人体生理学2006

    • Author(s)
      浦野哲盟
    • Total Pages
      11
    • Publisher
      朝倉書店
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary 2006 Annual Research Report
  • [Book] 血栓症ナビゲーター2006

    • Author(s)
      浦野哲盟
    • Total Pages
      2
    • Publisher
      メディカルレビュー社
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary 2006 Annual Research Report

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Published: 2006-04-01   Modified: 2016-04-21  

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