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Reconstitution of paracellular transport by establishment of a claudin-deficient epithelial cell line.

Research Project

Project/Area Number 16K15226
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field General medical chemistry
Research InstitutionNational Institute for Physiological Sciences

Principal Investigator

Furuse Mikio  生理学研究所, 生体機能調節研究領域, 教授 (90281089)

Research Collaborator OTANI Tetsuhisa  
SUGAWARA Taichi  
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Keywordsクローディン / タイトジャンクション / 上皮バリア機能 / 細胞間接着 / 上皮細胞 / ゲノム編集 / 傍細胞輸送
Outline of Final Research Achievements

We have established an epithelial cell line which lacks claudin-based tight junctions by disrupting claudin family genes, including claudin-2, claudin-4, claudin-3, claudin-7 and claudin-1, by genome editing in MDCK II cells. The cells, designated claudin quin KO cells, lacked claudin-based tight junction strands and showed the remarkably reduced epithelial barrier function with normal epithelial polarity. Introduction of each of the above claudins into claduin quin KO cells reconstituted functional tight junctions.

Academic Significance and Societal Importance of the Research Achievements

本研究の結果、特定のクローディンサブタイプだけで再構成されたタイトジャンクションの機能を解析することがはじめて可能となり、クローディンのサブタイプの機能特性の相違に基づく傍細胞経路の透過性制御の分子機構の理解が一層進む。さらに、本研究で世界に先駆けて樹立されたタイトジャンクション欠失上皮細胞の特性を解析することにより、上皮細胞におけるタイトジャンクションの意義を詳細に解析することが可能となった。その成果は上皮細胞生物学における細胞接着、細胞極性の研究領域で重要な知見となることが期待される。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report

Research Products

(7 results)

All 2019 2018 Other

All Journal Article (1 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (5 results) (of which Int'l Joint Research: 2 results) Remarks (1 results)

  • [Journal Article] Claudin-3-deficient C57BL/6J mice display intact brain barriers2019

    • Author(s)
      Castro DM、Coisne C、Lazarevic I、Baden P、Hata M、Iwamoto N、Francisco DMF、Vanlandewijck Mi、He L、Baier FA.、Stroka D、Bruggmann R、Lyck R、Enzmann G、Deutsch U、Betsholtz C、Furuse M、Tsukita Sh、Engelhardt B
    • Journal Title

      Scientific Reports

      Volume: 9 Pages: 36731-3

    • DOI

      10.1038/s41598-018-36731-3

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Establishment of a tight junction-deficient epithelial cell line by genome editing of claudin genes2019

    • Author(s)
      Mikio Furuse, Tetsuaki Otani, Taichi Sugawara, Shinsaku Tokuda, Mika Watanabe, Kyoko Furuse, Osamu Nagata
    • Organizer
      9th Federation of the Asian and Oceanian Physiological Sciences
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ZO family proteins regulate epithelial polarity independent of tight junction strand assembly2019

    • Author(s)
      Tetsuhisa Otani, Shinsaku Tokuda, Mikio Furuse
    • Organizer
      9th Federation of the Asian and Oceanian Physiological Sciences
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ZO family proteins regulate epithelial polarity independent of Tight Junction strand assembly2018

    • Author(s)
      Tetsuhisa Otani, Shinsaku Tokuda, Mikio Furuse
    • Organizer
      第70回日本細胞生物学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Angulin-1 regulates vertical elongation of tricellular tight junction by interacting with scaffolding proteins2018

    • Author(s)
      Taichi Sugawara, Mikio Furuse
    • Organizer
      第70回日本細胞生物学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] The role of tight junction-associated plaque proteins ZO-1 and ZO-2 in epithelial polarity2018

    • Author(s)
      古瀬幹夫、大谷哲久
    • Organizer
      第95回日本生理学会大会
    • Related Report
      2017 Research-status Report
  • [Remarks] 自然科学研究機構生理学研究所細胞構造研究部門

    • URL

      http://www.nips.ac.jp/dcs/index.html

    • Related Report
      2016 Research-status Report

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Published: 2016-04-21   Modified: 2020-03-30  

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