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機械受容機構の解明―メカノリセプターの分子実体は何か―

Research Project

Project/Area Number 11670037
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General physiology
Research InstitutionNagoya University

Principal Investigator

成瀬 恵治  名古屋大学, 医学部, 助教授 (40252233)

Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywords機械受容チャネル / 血管内皮細胞 / 細胞内カルシウム測定法 / クローニング / パッチクランプ / トリ心筋 / パッチクランプ法
Research Abstract

<HUVECのSAチャネルのクローニング>
HUVECよりcDNAライボラリーを作成し、発現ベクター(pcDNA3.1)に組込んだ.あらかじめ伸展依存性Caトランジェントがないことを確認したHEK細胞に25グループに分けた上記ライボラリーを遺伝子導入した。伸展可能シリコンチャンバー上に培養した遺伝子導入細胞にカルシウム蛍光色素FURA2を負荷し伸展刺激を与え、カルシウム上昇がある群を選んだ.現在、数群の伸展依存性Caトランジェントを起こすグループを同定した。
くトリ心筋SAチャネルのクローニング>
培養鶏胚心筋細胞には数種類のSAチャネルが存在する。このうちCa依存性Kチャネルの特徴を備えたSAチャネルがあるので、degenerate primerを作り、PCR産物を得,これを用いて12日鶏胚心筋細胞より得られたファージライボラリーをスクリーニングしたところORFが約3kbpのクローンを得た.これを発現ベクターに組込みCHO細胞に発現させたところ、培養鶏胚心筋細胞にパッチクランプをしたときに観察されたものと同じ性質のSAチャネルを観察する事が出来た。これは既知のイオンチャネルと数アミノ酸の相違があった。現在種々のミューテーションを加える事により機能-構造連関を解析しているところである。

Report

(3 results)
  • 2001 Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] Kada K: "Orientation Change of Cardiocytes Induced by Cyclic Stretch Stimulation : Time Dependency and Involvement of Protein Kinases"J. Mol. Cell. Cardio.. 31. 247-259 (1999)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] Numaguchi Y: "Prostacyclin synthase gene transfer accelerates reendothelializaton and inhibits neointimal formation in rat carotid arteries after balloon injury"Arteriosclerosis, Thrombosis, and Vascular Biology. 19. 723-733 (1999)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] Borgeld H: "Appropriately spaced nuclear localizing signals are necessary for nuclear import of non-nuclear proteins"Biophys. Biochem. Res. Com. 256. 278-283 (1999)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sai X: "Activation of pp60^<src> is critical for stretch-induced orienting response in fibroblasts"J. Cell Scienc. 112. 1365-1373 (1999)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kawakubo T: "Characterization of a newly found stretch-activated K_<ca,> ATP channel in cultured chick ventricular myocytes"Am. J. Physiol. 276. H1827-H1838 (1999)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kanzaki M: "Molecular identification of a eukaryotic, stretch-activated nonselective cation channel"Science. 285. 882-886 (1999)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] 曽我部正博: "細胞膜の変形と細胞応答-細胞の形が機能を決める?-"生体膜のダイナミクス. 7 (1999)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] 曽我部正博: "機械刺激による細胞のリモデリング"生体の科学. 9 (2000)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] Yamada T: "Stretch-induced morphological changes of human endothelial cells depend on the intracellular level of Ca^<2+> rather than of cAMP"Life Sciences. 67. 2605-2613 (2000)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] Naruse K: "Aldose reductase inhibition prevents glucose-induced apoptosis in cultured bovine retinal microvascular pericytes"Exp Eye Res. 71. 309-315 (2000)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] Goldmann W: "The Cytoskeleton and Signaling : A Practical Approach"Cell Shape Control and Mechanical Signalling through the Cytoskeleton. 31 (1999)

    • Related Report
      2001 Final Research Report Summary
  • [Publications] Yamada T: "Stretch-induced morphological changes of human endothelial cells depend on the intracellular level of Ca^<2+> rather than of cAMP"Life Sciences. 67. 2605-2613 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Naruse K: "Aldose reductase inhibition prevents glucose-induced apoptosis in cultured bovine retinal microvascular pericytes"Exp Eye Res. 71. 309-315 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 曽我部正博: "機械刺激による細胞のリモデリング"生体の科学. 9 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yamada T,Naruse K,and Sokabe M: "Stretch-induced morphological changes of human endothelial cells depend on the intracellular level of Ca^<2+> rather than of cAMP"Life Science. (in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kanzaki M,Sato C,Kojima I,Naruse K,Sokabe M,Iida H: "Molecular identification of a eukaryotic,stretch-activated nonselective cation channel"Science. 285. 882-886 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kawakubo T,Naruse K,Matsubara T,Hotta N,and Sokabe M: "Characterization of a newly found stretch-activated K_<Ca>,ATP channel in cultured chick ventricular myocytes."Am.J.Physiol.. 276. H1827-1838 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sai X,Naruse K,and Sokabe M: "Activation of pp60^<src> is critical for stretch-induced orienting response in fibroblasts."J.Cell Science. 112. 1365-1373 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Borgeld H,Naruse K,Nishikawa S,Zhang J,Kikuchi A,Furukawa K,Yagi: "Appropriately spaced nuclear localizing signals are necessary for nuclear import of non-nuclear proteins."Biophys.Biochem.Res.Com. 256. 278-283 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kada K,Yasui K,Naruse K,and Toyama J: "Orientation Change of Cardiocytes Induced by Cyclic Stretch Stimulation:Time Dependency and Involvement of Protein 1999 Kinases."J.Mol.Cell.Cardio.. 31. 247-259 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 曽我部正博、成瀬恵治: "生体膜のダイナミクス"細胞膜の変形と細胞応答-細胞の形が機能を決める?- (印刷中).

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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