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Molecular mechanisms of expression and regulation of function of water channel proteins aquaporins in the salivary gland cells

Research Project

Project/Area Number 11671844
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional basic dentistry
Research InstitutionUniversity of Tokushima

Principal Investigator

KAZUO Hosoi  Tokushima University School of Dentistry, Department of Physiology and Oral Physiology, Professor, 歯学部, 教授 (10049413)

Co-Investigator(Kenkyū-buntansha) AKAMATSU Tetsuya  University of Tokushima School of Dentistry, Department of Physiology and Oral Physiology, Research Associate, 歯学部, 助手 (80294700)
MATSUURA Sachiko  Matsumoto Dental University School of Dentistry Department of Histology, Assistant Professor, 歯学部, 講師 (50139854)
KANAMORI Norio  University of Tokushima School of Dentistry, Department of Physiology and Oral Physiology, Associate Professor, 歯学部, 助教授 (90064865)
TSUMURA Keiko  University of Tokushima School of Dentistry, Department of Physiology and Oral Physiology, Technical Official, 歯学部, 教務員 (50127841)
TADA Jun  University of Tokushima School of Dentistry, Department of Physiology and Oral Physiology, Research Associate, 歯学部, 助手 (00314865)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1999: ¥2,000,000 (Direct Cost: ¥2,000,000)
Keywordsaquaporins / trafficking / salivary gland / gastrointestinal tract / duodenum / Brunner's gland / 細胞骨格 / 細胞内カルシウムイオン
Research Abstract

AQPs are family of water channel proteins expressed in various tissues ; ten isoforms of the aquaporin family have been identified in mammals. AQP5 is an exocrine type water channel strongly expressed in the salivary gland, lacrimal gland, and lung, and is thought to play a fundamental role in water movement in formation of the saliva, tears, and other secretions. On the other hand, there are only few reports about its trafficking, water transport, or gene regulation . The purpose of the present study is therefore to understand the mechanism of AQP5 trafficking from the cytosol to the plasma membrane. Here we established AQP5-gene-transfected HSG cells since no AQP5-expressing cell line that has originated from the salivary glands was available.
A cDNA of rat aquaporin 5 (AQP5) was used to transfect to HSG (human salivary gland cells), and the trafficking mechanism was studied in vitro by confocal laser microscopy. The trafficking of AQP5 to the plasma membrane was induced by stimulatio … More n of AQP5-gene-transfected human salivary gland cells with thapsigargin, an inhibitor of endoplasmic Ca^<2+>-ATPase, or with A-23187, a calcium ionophore. Pretreatment of these cells with colchicine or vinblastine, microtubule inhibitors, prevented the trafficking induced by thapsigargin or A-23187. The trafficking event was not completely inhibited by cytochalasin B, a microfilament inhibitor. These results demonstrate that the trafficking of AQP5 vesicles to the plasma membrane is triggered by an increase in intracellular Ca^<2+> and that the interaction of AQP5-containing vesicles with the cytoskeleton is involved in this trafficking.
We also investigated the expression and localization of AQP5 in the gastrointestinal tract of the rat, since this tract transports a large amount of water and is mainly composed of many exocrine tissues, we tried to detect the expression and localization of AQP5 in this tract.
Aquaporin 5 (AQP5) mRNA was detected in the lower stomach and duodenum by reverse transcriptase-polymerase chain reaction (RT-PCR). RT-PCR Southern blotting demonstrated the presence of AQP5 in other tissues of the gastrointestinal tract i.e., upper stomach, lower stomach, duodenum, ileum, caecum, and colon but not in jejunum, and rectum. Other AQPs i.e., AQP1, AQP3, and AQP4 mRNAs were also detected in the lower stomach and duodenum by RT-PCR.Western blot analysis detected the AQP5 protein in the lower stomach and duodenum. Immunohistochemical analysis demonstrated that AQP5 was localized in the apical and lateral membrane of the Brunner's gland in the duodenum. These results suggest that AQPs are functional in the fluid secretion or absorption in the gastrointestinal tract. Less

Report

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

    (44 results)

All Other

All Publications (44 results)

  • [Publications] Kurabuchi,S. et al: "An unusual sexually dimorphic mosaic distribution of a subset of kallikreins in the granular convoluted tubule of the mouse submandibular galnd detected by an antibody with restricted immunoreactivity"Histochemical Journal. 31. 19-28 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Akamatsu,T. et al: "Subtilisin-like proprotein convertase PACE4 (SPC4) is a cndidate processing enzyme of bone morphogenic protein during tooth formation"Developmental Dynamics,. 216. 481-488 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sakai,K.and Kanamori,N.: "Are there non-aminergic paradoxical sleep-off neurons in the brainstem?"Sleep Res.Online. 2. 57-63 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tada,J. et al: "Involvement of vesicle-cytoskeleton interaction in AQP5 trafficking in AQP5-gene-transfected HSG cells,"Biochemical and Biophysical Research Communications. 266(2). 443-447 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 市川哲雄 他: "高齢者における口腔ケアと健康に関する基礎的研究"日本歯科医学誌. 19. 75-80 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Akamatus,T. et al: "Highly regulated expression of subtilisin-like proprotein convertase PACE4 (SPC4) during dentinogenesis"Biochemical and Biophysical Research Communications. 272. 410-415 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hosoi,K. et al: "Expression of kininogens in the connective tissue-type mast cells of the rat"Immunology. 101(4). 531-540 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kanamori,N. et al: "Search for the Dye that Stains the Vascular System"TE Lett.Batt.New Tech.Med.. 1(in press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Wei,W. et al: "Induction of C-reactive protein, serum amyloid P component, and kininogens in the submandibular and lacrimal glands of rats with experimentally induced inflammation"Life Sciences. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Parvin,M.N. et al: "Structure, function, and transcriptional regulation of aquaporins, proteins of the water channel family"Dentistry in Japan. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 倉淵真悟 他: "マウス顎下腺顆粒性導管の性差とホルモンによる調節-免疫組織化学的解析-"日本咀嚼学会雑誌. (印刷中). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Parvin,M.N., et al: "Expression and hormone-induced redistribution of AQP5 in Brunner's gland of the rat duodenum"Biochimica et Biophysica Acta. (in submission).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kurabuchi,S. et al: "Androgen regulation of the cellular distribution of the true tissue kallikrein, mk1, in the submandibular gland of the mouse"Journal of Histochemistry and Cytochemistry. (in submission).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takaishi,K. et al: "Effects of local anesthetics on L-arginine transport in cultured bovine aortic endothelial cells."Anesthesia and Analgia. (in submission).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kurabuchi,S. et al: "Action of androgens on the immunocytochemical distribution of mK1, a true tissue kallikrein, in the granular convoluted tubule of the mouse submandibular gland"(in preparation).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 細井和雄(分担執筆)他: "分子細胞生物学基礎実験法 改訂第2版"南江堂(印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kurabuchi, S., Tada, J., Gresik, E.W., and Hosoi, K.: "An unusual sexually dimorphic mosaic distribution of a subset of kallikreins in the granular convoluted tubule of the mouse submandibular galnd detected by an antibody with restricted immunoreactivity"Histochemical Journal. 31(1). 19-28 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Akamatsu, T., Matsuda, Y., Tsumura, K., Tada, J., Parvin, M.N., Kanamori, N., Hosoi, K.: "Subtilisin-like proprotein convertase PACE4 (SPC4) is a cndidate processing enzyme of bone morphogenic protein during tooth formation"Developmental Dynamics. 216(4/5). 481-488 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sakai, K., and Kanamori, N.: "Are there non-aminergic paradoxical sleep-off neurons in the brainstem?"Sleep Research on line. 2. 57-63

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tada, J., T.Sawa, N.Yamanaka, M.Shono, T.Akamatsu, K.Tsumura, M.N.Parvin, N.Kanamori, and K.Hosoi: "Involvement of vesicle-cytoskeleton interaction in AQP5 trafficking in AQP5-gene-transfected HSG cells"Biochemical and Biophysical Research Communications. 266(2). 443-447 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ichikawa, T., Y.Terada, K.Hirota, Y.Miyake, and K.Hosoi: "Study on oral health care and general health of elderly (in Japanese)"Journal of the Japanese Association for Dental Science. 19. 75-80 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Akamatus, T., Matusda, Y., Tsumura, K., Tada, J., Parvin, M.N., Wei, W., Kanamori, N., and Hosoi, K.: "Highly regulated expression of subtilisin-like proprotein convertase PACE4 (SPC4) during dentinogenesis"Biochemical and Biophysical Research Communications. 272(2). 410-415 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hosoi, K., S.Matsuura, K.Tsumura, W.Wei, M.N.Parvin, J.Tada, T.Akamatsu, and K.Suzuki: "Expression of kininogens in the connective tissue-type mast cells of the rat"Immunology. 101(4). 531-540 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kanamori, N., Akamatsu, T., Tada, J., Tsumura, K., Parvin, M.N., Murdiastuti, K., Wei, W., and Hosoi, K.: "Search for the Dye that Stains the Vascular System"ITE Lett.Batt.New Tech.Med.. Vol.1, No.4 (in press). (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Wei, W., M.N.Parvin, K.Tsumura, T.Akamatsu, J.Tada, N.Kanamori, and K.Hosoi: "Induction of C-reactive protein, serum amyloid P component, and kininogens in the submandibular and lacrimal glands of rats with experimentally induced inflammation"Life Sciences. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Parvin, M.N., J.Tada, T.Akamatsu, and K.Hosoi: "Structure, function, and transcriptional regulation of aquaporins, proteins of the water channel family"Dentistry in Japan. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kurabuchi, S., and Hosoi, K.: "Sexual dimorphism and hormonal regulation of granular convoluted tubule (GCT) cells of mouse submandibular gland-Immunohistochemical analysis- (In Japanese)"Journal of Japanese Society for mastication Science and Health Promotion. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Parvin, M.N., K.Tsumura, W.Wei, J.Tada, T.Akamatsu, N.Kanamori, and K.Hosoi: "Expression and hormone-induced redistribution of AQP5 in Brunner's gland of the rat duodenum"Biochimica et Biophysica Acta. (submitted).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kurabuchi, S.K.Hosoi, and E.W.Gresik: "Androgen regulation of the cellular distribution of the true tissue kallikrein, mk1, in the submandibular gland of the mouse"Journal of Histochemistry and Cytochemistry. (submitted).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takaishi, K., S.Tomioka, N.Nakajp, and K.Hosoi: "Effects of local anesthetics on L-arginine transport in cultured bovine aortic endothelial cells"Anesthesia and Analgia. (submitted).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kurabuchi, S., K.Hosoi, and E.W.Gresik: "Action of androgens on the immunocytochemical distribution of mK1, a true tissue kallikrein, in the granular convoluted tubule of the mouse submandibular gland"Journal of Histochemistry and Cytochemistry. (in submited).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Hosoi et al: "Experimental Manual for Molecular Cell Biology, 2nd Edition (in Japanese), edited by T.Kakuno, K.Nishikawa, T.Higuti, Y.Horiuchi, Y.Matuo, K.Miyazaki, K.Hosoi, and N.Aragaki (compiled under the supervision of T.Horio)"Nankodo Publisher (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tada,J. et al: "Involvement of vesicle-cytoskeleton interaction in AQP5 trafficking in AQP5-gene-transfected HSG cells,"Biochemical and Biophysical Research Communications. 266(2). 443-447 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 市川哲雄 他: "高齢者における口腔ケアと健康に関する基礎的研究"日本歯科医学誌. 19. 75-80 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Akamatus,T. et al: "Highly regulated expression of subtilisin-like proprotein convertase PACE4 (SPC4) during dentinogenesis"Biochemical and Biophysical Research Communications. 272. 410-415 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hosoi,K. et al: "Expression of kininogens in the connective tissue-type mast cells of the rat"Immunology. 101(4). 531-540 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kanamori,N. et al: "Search for the Dye that Stains the Vascular System"TE Lett.Batt.New Tech.Med.. 1(in press). (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Wei,W. et al: "Induction of C-reactive protein, serum amyloid P component, and kininogens in the submandibular and lacrimal glands of rats with experimentally induced inflammation"Life Sciences. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Parvin,M.N. et al: "Structure, function, and transcriptional regulation of aquaporins, proteins of the water channel family"Dentistry in Japan. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 倉淵真悟 他: "マウス顎下腺顆粒性導管の性差とホルモンによる調節 -免疫組織化学的解析-"日本咀嚼学会雑誌. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 細井和雄(分担執筆) 他: "分子細胞生物学基礎実験法 改訂第2版"南江堂 (印刷中).

    • Related Report
      2000 Annual Research Report
  • [Publications] Kurabuchi,S.et al: "An unusual sexually dimorphic mosaicdistribution of a subset of kallikreins in the granular convoluted tubule of the mouse submandibalar gland detected by an antibody with restricted immunoreactivity"Histochemical Journal. 31(1). 19-28 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Akamatsu,T.et al: "Subtilisin-like proprotein convertase(SPC4) is a candidate processing enzyme of bone morphogenic protein during tooth formation"Developmental Dynamics. 216(4/5). 481-488 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Tada,J.et al: "Involvement of vesicle-cytoskelton interaction in AQP5 trafficking in AQP5-gene-transfected HSG cells"Biochemical and Biophysical Research Communications. 266(2). 443-448 (1999)

    • Related Report
      1999 Annual Research Report

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

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