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Structural and functional analyses of E-face-associated tight junction strands formed by expression of claudin-1 mutant in the second extracellular loop in MDCKII cells

Research Project

Project/Area Number 18590187
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General anatomy (including Histology/Embryology)
Research InstitutionKyushu University

Principal Investigator

INAI Tetsuichiro  Kyushu University, Department of Developmental Molecular Anatomy, Graduate School of Medical Sciences, Associate Professor (00264044)

Co-Investigator(Kenkyū-buntansha) HIROSE Eiji  KYUSHU UNIVERSITY, Department of Developmental Molecular Anatomy, Graduate School of Medical Sciences, Assistant Professor (40380620)
SHIBATA Yosaburo  KYUSHU UNIVERSITY, Department of Developmental Molecular Anatomy, Graduate School of Medical Sciences, Professor (90037482)
Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥300,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2006: ¥2,600,000 (Direct Cost: ¥2,600,000)
Keywordstight junction / claudin / freeze fracture / MDCK cells / paracellular permeability / タイト結合 / フリーズフラクチャー / MDCK細胞
Research Abstract

When epithelial cells are observed by freeze-fracture electron microscopy, continuous tight junction strands consisted of intramembranous particles and complementary grooves without particles are observed in the protoplasmic (P)-face and exoplasmic (E)-face, respectively (P-type tight junction). In contrast, tight junction particles of endothelial cells of peripheral venules are located on the grooves in the E-face (E-type tight junction). Claudins are integral membrane proteins at tight junctions, consist of at least 24 members, and reconstitute tight junction strands in fibroblasts without occludin. Claudins consist of four transmembrane domains with both NH2-and COOH-termini located in the cytoplasm, two extracellular loops (ECL1 and ECL2), and one cytoplasmic loop. It is thought that structure and function of tight junctions may be determined by combination and mixing ratio of claudin species expressed in the tight junction. We constructed expression vectors containing claudin-1 wi … More th a myc-epitope at the COOH-terminus (1CLmyc), 1CLmyc deleted two amino acids (151P152L) in the ECL2(1CLAPLmyc), and claudin-10 with a myc-epitope at the COOH-terminus (10CLmyc), and claudin-10 itself. We then expressed them in MDCK I or II cells. When 1CLmyc-expressing cells were observed by freeze-fracture electron microscopy, P-type tight junctions were detected in lateral plasma membranes. 10CLmyc formed E-type tight junction in lateral membranes. However, 1CLAPLmyc formed intermediate type of tight junction whose intramembranous particles were associated equally with the P- and E-face. Transepithelial electrical resistances of 1CLmyc-and 1CLAPLmyc-expressing cells were significantly increased, but that of claudin-10 was significantly decreased. These results suggest that the ECL2 of claudin-1 is involved in determining whether tight junction particles associate with the P- or E-face, and that P-type and intermediate-type tight junctions increased transepithelial electrical resistance but E-type tight junction decreased transepithelial electrical resistance. Less

Report

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

    (33 results)

All 2008 2007 2006 Other

All Journal Article (20 results) (of which Peer Reviewed: 8 results) Presentation (13 results)

  • [Journal Article] Formation of aberrant TJ strands by overexpression of claudin-15 in MDCK II cells2008

    • Author(s)
      Sengoku A, Inai T, Shibata Y.
    • Journal Title

      Histochem Cell Biol 129(2)

      Pages: 211-22

    • 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] Comparative characterization of mouse rectum CMT93-I and-II cells by expression of claudin isoforms and tight junction morphology and function2008

    • Author(s)
      Inai T, Sengoku A, Hirose E, Iida H, Shibata Y
    • Journal Title

      Histochem Cell Biol 129

      Pages: 223-323

    • 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] Tektin5, a new Tektin family member, is a component of the middle piece of flagella in rat spermatozoa2008

    • Author(s)
      Murayama, E., Yamamoto, E., Kaneko, T., Shibata, T., Inai, T., Iida, H
    • Journal Title

      Mol Reprod Dev 75(4)

      Pages: 650-658

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Formation of aberrant TJ strands by overexpression of claudin-15 in MDCK II cells2008

    • Author(s)
      Sengoku, A., Inai, H., Shibata, Y
    • Journal Title

      Histochem Cell Biol 129(2)

      Pages: 211-222

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Comparative characterization of mouse rectum CMT93-I and -II cells by expressio of claudin isoforms and tight junction morphology and function2008

    • Author(s)
      Inai, T., Sengoku, A., Hirose, E., Iida, H., Shibata, Y
    • Journal Title

      Histochem Cell Biol 129(2)

      Pages: 223-232

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] タイト結合の構造と機能(総説)2008

    • Author(s)
      稲井, 哲一朗, 柴田, 洋三郎
    • Journal Title

      福岡医誌 99(2)

      Pages: 25-31

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Formation of aberrant TJ strands by overexpression of claudin-15 in MDCK II cells2008

    • Author(s)
      Sengoku A, Inai T, Shibata Y.
    • Journal Title

      Histochem Cell Biol. 129(2)

      Pages: 211-22

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Comparative characterization of mouse rectum CMT93-I and-II cells byexpression of claudin isoforms and tight junction morphology and function2008

    • Author(s)
      Inai T, Sengoku A, Hirose E, Iida H, Shibata Y
    • Journal Title

      Histochem Cell Biol. 129

      Pages: 223-323

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Claudin-7 Expressed on Lateral Membrane of Rat Epididymal Epithelium does not Form Aberrant Tight Junction Strands2007

    • Author(s)
      Inai T, Sengoku A, Hirose E, Iida H, Shibata Y
    • Journal Title

      Anat Rec (Hoboken) 290(11)

      Pages: 1431-8

    • 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] Germ cell-less like 2 protein is a new component of outer dense fibers in rat sperm flagella2007

    • Author(s)
      Murayama E, Katoh M, Kanebayashi A, Kaneko T, Shibata Y, Inai T, Iida H
    • Journal Title

      Reproduction 134(6)

      Pages: 749-56

    • 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] Claudin-7 expressed on lateral membrane of rat epididymal epithelium does not form aberrant tight junction strands2007

    • Author(s)
      Inai, T., Sengoku, A., Hirose, E., Iida, H., Shibata, Y
    • Journal Title

      Anat Rec 290(11)

      Pages: 1431-1438

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Germ cell-less like 2 protein is a new component of outer dense fibers in rat sperm flagella2007

    • Author(s)
      Murayama, E., Katoh, M., Kanebayashi, A., Kaneko, T., Shibata, Y., Inai, T., Iida, H
    • Journal Title

      Reproduction 134(6)

      Pages: 749-756

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] : Claudin-7 Expressed on Lateral Membrane of Rat Epididymal Epithelium does not Form Aberrant Tight Junction Strands2007

    • Author(s)
      Inai T, Sengoku A, Hirose E, Iida H, Shibata Y
    • Journal Title

      Anat Rec (Hoboken). 290(11)

      Pages: 1431-8

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Germ cell-less like 2 protein is a new component of outer dense fibers in rat sperm flagella2007

    • Author(s)
      Murayama E, Katoh M, Kanebayashi A, Kaneko T, Shibata Y, Inai T, Iida H
    • Journal Title

      Reproduction. 134(6)

      Pages: 749-56

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Spetex-1 : a new component in the middle piece of flagella of rodent spermatozoa2006

    • Author(s)
      Iida, H., Honnda, T., Matsuyama, Y., Shibata, Inai, T
    • Journal Title

      Mol Reprod Dev 73(3)

      Pages: 342-349

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Tektin 4 is located on outer dense fibers, not associated with axonemal tubulinsof flagella in rodent spermatozoa2006

    • Author(s)
      Iida, H., Honnda, T., Matsuyama, Y., Shibata, Inai, T
    • Journal Title

      Mol Reprod Dev 73(7)

      Pages: 929-936

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Rapid vascular regrowth in tumors after reversal of VEGF inhibition2006

    • Author(s)
      Mancuso, MR, Davis, R., Norberg, SM., O'Brien, S., Sennino, B., Nakahara, T., Yao, VJ., Inai, T, Brooks, P., Freimark, B., Shalinsky, DR., Hu-Lowe, DD, McDonald, DM
    • Journal Title

      J Clin Invest 116(10)

      Pages: 2610-2621

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Tektin 4 is located on outer dense fibers, not associated with axonemal tubulins of flagella in rodent spermatozoa.2006

    • Author(s)
      Iida H, Honda Y, Matsuyama T, Shibata Y, Inai T
    • Journal Title

      Mol Reprod Dev 73

      Pages: 929-936

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Rapid vascular regrowth in tumors after reversal of VEGF inhibition.2006

    • Author(s)
      Mancuso MR, Davis R, Norberg SM, O' Brien S, Sennino B, Nakahara T, Yao VJ, Inai T, Brooks P, Freimark B, Shalinsky DR, Hu-Lowe DD, McDonald DM
    • Journal Title

      J Clin Invest 116

      Pages: 2610-2621

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Differential expression of the tight junction proteins, claudin-1, claudin-4, occludin, ZO-1, and PAR3, in the ameloblasts of rat upper incisors

    • Author(s)
      Inai, T., Sengoku, A., Hirose, E., Iida, H., Shibata, Y
    • Journal Title

      Anat Rec 291(5)

      Pages: 577-585

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] タイト結合蛋白claudinによるタイト結合の形態と細胞間透過性の制御(シンポジウム)2008

    • Author(s)
      稲井 哲一朗、柴田 洋三郎
    • Organizer
      第113回 日本解剖学会総会・全国学術集会
    • Place of Presentation
      大分大学医学部
    • Year and Date
      2008-03-28
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] タイト結合蛋白claudinによるタイト結合の形態と細胞間透過性の制御(シンポジウム)2008

    • Author(s)
      稲井 哲一朗、柴田 洋三郎
    • Organizer
      第113回日本解剖学会総会・全国学術集会
    • Place of Presentation
      大分大学医学部
    • Year and Date
      2008-03-28
    • Related Report
      2007 Annual Research Report
  • [Presentation] 血管新生阻害剤による腫瘍血管の退縮過程:組織学的変化を追う(シンポジウム)2008

    • Author(s)
      稲井 哲一朗、柴田 洋三郎
    • Organizer
      第113回 日本解剖学会総会・全国学術集会
    • Place of Presentation
      大分大学医学部
    • Year and Date
      2008-03-27
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] P1-084 claudin-1の細胞外第二ループの変異によるタイト結合の形態変化の観察(ポスター)2008

    • Author(s)
      仙石 昭仁、稲井 哲一朗、廣瀬 英司、柴田 洋三郎
    • Organizer
      第113回 日本解剖学会総会・全国学術集会
    • Place of Presentation
      大分大学医学部
    • Year and Date
      2008-03-27
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 血管新生阻害剤による腫瘍血管の退縮過程:組織学的変化を追う(シンポジウム)2008

    • Author(s)
      稲井 哲一朗、柴田 洋三郎
    • Organizer
      第113回日本解剖学会総会・全国学術集会
    • Place of Presentation
      大分大学医学部
    • Year and Date
      2008-03-27
    • Related Report
      2007 Annual Research Report
  • [Presentation] P1-084 claudin-1の細胞外第二ループの変異によるタイト結合の形態変化の観察(ポスター)2008

    • Author(s)
      仙石 昭仁、稲井 哲一朗、廣瀬 英司、柴田 洋三郎
    • Organizer
      第113回日本解剖学会総会・全国学術集会
    • Place of Presentation
      大分大学医学部
    • Year and Date
      2008-03-27
    • Related Report
      2007 Annual Research Report
  • [Presentation] Regression process of tumor blood vessels treated with anti-angiogenic agents(symposium)2008

    • Author(s)
      T., Inai, Y., Shibata
    • Organizer
      The 113rd Annual Meeting of Japanese Association of Anatomists
    • Place of Presentation
      Oita University
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Regulation of tight junction structure and paracellular permeability by claudins(symposium)2008

    • Author(s)
      T., Inai, A., Sengoku, Y., Shibata
    • Organizer
      The 113rd Annual Meeting of Japanese Association of Anatomists
    • Place of Presentation
      Oita University
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Structural changes of tight junctions induced by claudin-1 mutations in the second extracellular loop2008

    • Author(s)
      A., Sengoku, T., Inai, E., Hirose, Y., Shibata
    • Organizer
      The 113rd Annual Meeting of Japanese Association of Anatomists
    • Place of Presentation
      Oita University
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Freeze-fracture electron microscopy of ectopic tight junction strands induced by expression of claudin-15 in MDCK II cells2007

    • Author(s)
      A., Sengoku, T., Inai, E., Hirose, Y., Shibata
    • Organizer
      The 112nd Annual Meeting of Japanese Association of Anatomists
    • Place of Presentation
      Osaka University
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] New cell lines established from CMT93 cells, mouse rectum carcinoma cells, for investigaton of tight junctions2007

    • Author(s)
      T., Inai, A., Sengoku, E., Hirose, Y., Shibata
    • Organizer
      The 112nd Annual Meeting of Japanese Association of Anatomists
    • Place of Presentation
      Osaka University
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Immunohistochemical and immunocytochemical localization of GATE-16 which binds to microtube regulatory factor, RB32006

    • Author(s)
      E., Hirose, H., Morii, Y., Naruse, T., Indi, Y., Shibata, N., Mori
    • Organizer
      The 62th Annual Meeting of Japanese Association of Anatomists(Kyushu branch)
    • Place of Presentation
      Kyushu Universit
    • Year and Date
      2006-10-21
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Connexin Diversity and Cardiac Conduction Development2006

    • Author(s)
      Y., Shibata, K., Nishii, T., Inai, M., Kumai
    • Organizer
      The 16th International Microscopy Congress
    • Place of Presentation
      Sapporo
    • Year and Date
      2006-09-07
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary

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

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