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Physiological function and localization of a novel rCLCA molecule related to Cl-flux in salivary tissues

Research Project

Project/Area Number 17591958
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional basic dentistry
Research InstitutionFukuoka Dental College

Principal Investigator

YAMAZAKI Jun  Fukuoka Dental College, School of Dentistry, Assoc. Prof. (50230397)

Co-Investigator(Kenkyū-buntansha) OKAMAURA Kazuhiko  Fukuoka Dental College, School of Dentistry, Assoc. Prof. (00224056)
ISHIBASHI Kazunari  Fukuoka Dental College, School of Dentistry, Lecturer (50122768)
Project Period (FY) 2005 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,650,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥150,000)
Fiscal Year 2007: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2006: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2005: ¥2,300,000 (Direct Cost: ¥2,300,000)
KeywordsC1^- channels / salivary tissues / duct / epithelium / endosome / transporting vesicle / saliva / post-translational modification / CLCA / Rab / Ca^<2+>活性化Cl^-チャネル / 細胞内小器官 / エンドソ-ム / Cl^-輸送 / 顎下腺 / 唾液分泌 / N型糖鎖 / 口腔乾燥症
Research Abstract

Epithelial Cl^- transport is known to be important for saliva production although a molecular entity for Ca^<2+>-dependent C1^- transport has not been well characterized. We identified a new member of the CLCA family, rCLCA, which is strongly expressed in the submandibular gland (SMG). The predicted ORF encodes a 903-a.a. protein containing an N-terminal region located extracellularly. Transient and stable transfection of HEK293 cells with rCLCA cDNA resulted in expression of 120^- and 86-kDa proteins in the membrane fraction. The latter appeared to be a proteolytic product of the former. Streptavidin recognized the 86-kDa protein immunoprecipitated from surface-biotinylated cells using the anti-rCLCA antibody, suggesting that the proteolytic product locates on the plasma membrane. The transfection with rCLCA revealed a marked niflumic acid-sensitive Ca^<2+>-dependent Cl^- conductance evoked by ionomycin with an outward rectification in the I-V relationship. This molecule was also found to be expressed in early and recycling endosomes labeled by their specific markers. RT-PCR revealed expression of rCLCA in the stomach, lung and liver as well as the SMG. Intense immunostaining was detected in the striated ducts of SMG, but not in the acinar cells. Immunoblot of the membrane fraction yielded N-glycan-linked 120-130- and 90-kDa bands. To address the question as to whether rCLCA is involved in Cl^- reabsorption in rat SMG during muscarinic stimulation, we evaluated the electrolyte content of saliva from the glands pretreated with rCLCA siRNA. Retrograde injection into a given submandibular duct with the siRNA significantly increased Cl^- concentration in saliva collected after administration of pilocarpine. These results represent a physiological significance of rCLCA in transepithelial Ca^<2+>-dependent Cl^- transport in the ductal system of rat SMG.

Report

(4 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • Research Products

    (28 results)

All 2008 2007 2006 2005

All Journal Article (9 results) (of which Peer Reviewed: 4 results) Presentation (17 results) Book (2 results)

  • [Journal Article] Involvement of apical P2Y_2 receptor-regulated CFTR activity in muscarinic stimulation of Cl^- reabsorption in rat submandibular gland2008

    • Author(s)
      Ishibashi, K., Okamura, K., and Yamazaki, J.
    • Journal Title

      Am.J.Physiol.:Regul.Integr.Comp.Physiol 294

    • 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] Involvement of apical P2Y_2 receptor-regulated CFTR activity in muscarinic stimulation of C1 reabsorption in rat submandibular gland2008

    • Author(s)
      Ishibashi, K., Okamura. K., and Yamazaki, J.
    • Journal Title

      Am. J. Physiol. : Regul. Integr. Comp. Physiol. 294

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Involvement of apical P2Y_2 receptor-regulated CFTR activity in muscarinic stimulation of Cl rcabsortion in rat submandibular gland2008

    • Author(s)
      Ishibashi, K., Okamura, K., and Yamazaki, J.
    • Journal Title

      Am.J.Physiol.:Regul.Integr.Comp.Physiol 294

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Roles of CLCA and CFTR in electrolyte re-absorption from rat saliva2006

    • Author(s)
      Ishibashi, K., Yamazaki, J., Okamura, K., Teng, Y., Kitamura, K. and Abe, K.
    • Journal Title

      J.Dent.Res. 85

      Pages: 1101-1105

    • 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] Roles of CLCA and CFTR in electrolyte re-absorption from rat saliva2006

    • Author(s)
      Ishibashi. K.,Yamazaki, J., Okamura, K., Teng, Y., Kitamura, K., Abe, K.
    • Journal Title

      J. Dent. Res. 85

      Pages: 1101-1105

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Roles of CLCA and CFTR in electrolyte re-absorption from rat saliva2006

    • Author(s)
      Ishibashi, K., Yamazaki, J., Okamura, K., Teng, Y., Kitamura, K., Abe, K.
    • Journal Title

      J.Dent.Res. 85(12)

      Pages: 1101-1105

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Characterization of CLCA protein expressed in ductal cells of rat salivary glands2005

    • Author(s)
      Yamazaki, J., Okamura, K., Ishibashi, K. and Kitamura K.
    • Journal Title

      Biochim.Biophys.Acta.-Biomemb. 1715

      Pages: 132-144

    • 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] Characterization of CLCA protein expressed in ductal cells of rat salivary glands2005

    • Author(s)
      Yamazaki, J., Okamura, K., Ishibashi, K., Kitamura, K.
    • Journal Title

      Biochim. Biophys. Acta. -Biomemb. 1715

      Pages: 132-144

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Characterization of CLCA protein expressed in ductal cells of rat salivary glands2005

    • Author(s)
      Yamazaki, J., Okamura, K., Ishibashi, K., Kitamura K.
    • Journal Title

      Biochim.Biophys.Acta.-Biomemb. 1715

      Pages: 132-144

    • Related Report
      2005 Annual Research Report
  • [Presentation] CLCAスプライシング変異型の細胞特異的な発現と細胞接着に対する抑制作用2008

    • Author(s)
      山崎 純, 岡村 和彦, 他
    • Organizer
      第81回 日本薬理学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2008-03-19
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] CLCAスプライシング変異型の細胞特異的な発現と細胞接着に対する抑制作用2008

    • Author(s)
      山崎 純、岡村 和彦, 他
    • Organizer
      第81回日本薬理学会年会
    • Place of Presentation
      横浜
    • Year and Date
      2008-03-19
    • Related Report
      2007 Annual Research Report
  • [Presentation] ラット顎下腺における CLCAアイソフォームの選択的発現と機能2007

    • Author(s)
      山崎 純, 岡村 和彦
    • Organizer
      第52回 日本唾液腺学会
    • Place of Presentation
      東京
    • Year and Date
      2007-12-08
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ラット顎下腺におけるCLCAアイソフォームの選択的発現と機能2007

    • Author(s)
      山崎 純、岡村 和彦
    • Organizer
      第52回日本唾液腺学会
    • Place of Presentation
      東京
    • Year and Date
      2007-12-08
    • Related Report
      2007 Annual Research Report
  • [Presentation] cAMP活性化Cl^-チャネルCFTRによるラット顎下腺唾液イオン濃度の制御2007

    • Author(s)
      石橋 一成, 山崎 純, 他
    • Organizer
      第49回 歯科基礎医学会学術大会
    • Place of Presentation
      札幌
    • Year and Date
      2007-08-30
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] cAMP活性化Cl-チャネルCFTRによるラット顎下腺唾液イオン濃度の制御2007

    • Author(s)
      石橋 一成、山崎 純, 他
    • Organizer
      第49回歯科基礎医学会学術大会
    • Place of Presentation
      札幌
    • Year and Date
      2007-08-30
    • Related Report
      2007 Annual Research Report
  • [Presentation] 唾液腺におけるCl^-チャネル関連分子CLCAの局在と機能2007

    • Author(s)
      山崎 純
    • Organizer
      第49回 歯科基礎医学会学術大会サテライトシンポジウム
    • Place of Presentation
      札幌
    • Year and Date
      2007-08-29
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 唾液腺におけるCl-チャネル関連分子CLCAの局在と機能2007

    • Author(s)
      山崎 純
    • Organizer
      第49回歯科基礎医学会学術大会サテライトシンポジウム
    • Place of Presentation
      札幌
    • Year and Date
      2007-08-29
    • Related Report
      2007 Annual Research Report
  • [Presentation] 翻訳後修飾を受けたCa^<2+>活性化Cl^-チャネル関連分子の細胞内局在の差異2007

    • Author(s)
      山崎 純, 岡村 和彦, 北村 憲司
    • Organizer
      第80回 日本薬理学会年会
    • Place of Presentation
      名古屋
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Localization and Function of CLCA in rat submandibular glands2006

    • Author(s)
      Jun Yamazaki.
    • Organizer
      3rd International Symposium on Salivary Glands in Honor of Niels Stensen
    • Place of Presentation
      Okazaki,Japan
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ラット顎下腺に局在するCa^<2+>活性化Cl^-チャネル関連分子CLCAの性質2006

    • Author(s)
      山崎 純, 岡村 和彦, 他
    • Organizer
      第48回 歯科基礎医学会学術大会
    • Place of Presentation
      鶴見
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ラットCa^<2+>活性化C1^-チャネル関連分子rCLCAの局在2006

    • Author(s)
      岡村 和彦, 山崎 純, 他
    • Organizer
      第48回 歯科基礎医学会学術大会
    • Place of Presentation
      鶴見
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ラット顎下腺導管上皮細胞における再吸収においてClイオンチャネルの果たす役割2006

    • Author(s)
      石橋 一成, 山崎 純, 他
    • Organizer
      第48回 歯科基礎医学会学術大会
    • Place of Presentation
      鶴見
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] ラット顎下腺導管におけるCl^-チャネル関連分子CLCAアイソフォームの局在と機能2006

    • Author(s)
      山崎 純, 岡村 和彦, 他
    • Organizer
      第83回 日本生理学会大会
    • Place of Presentation
      群馬
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 短縮型CLCAの分子クローニングとラット顎下腺における発現2006

    • Author(s)
      山崎 純, 北村 憲司
    • Organizer
      第79回 日本薬理学会年会
    • Place of Presentation
      横浜
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Cloning and Characterization of CLCA Protein Expressed in Ductal Cells of Rat Salivary Glands2006

    • Author(s)
      Jun Yamazaki, Kazuhiko Okamura, Kazunari Ishibashi, et. al.
    • Organizer
      50th Meeting of Biophysical Society
    • Place of Presentation
      Salt Lake City,Utah,USA
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Ca^<2+>活性化Cl^-チャネル関連分子CLCAのラット唾液腺における局在と機能2005

    • Author(s)
      山崎 純, 石橋 一成, 他
    • Organizer
      第50回 日本唾液腺学会
    • 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] イオンチャネル創薬-現状とその可能性 7.Cl^-チャネル創薬の現状と展望, 医学のあゆみ223(6)2007

    • Author(s)
      山崎 純
    • Publisher
      医歯薬出版
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Book] イオンチャネル創薬-現状とその可能性7.Clチャネル創薬の現状と展望、医学のあゆみ223(6)2007

    • Author(s)
      山崎 純
    • Publisher
      医歯薬出版
    • Related Report
      2007 Annual Research Report

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Published: 2005-04-01   Modified: 2016-04-21  

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