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Identification of molecules which decide the region where the exocytosis occurs, and molecular mechanisms for exocytotic membrane fusion in salivary gland

Research Project

Project/Area Number 05454508
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Functional basic dentistry
Research InstitutionAsahi University

Principal Investigator

KAMEYAMA Yasunaga  Asahi univ. School of Dentistry, Associate Prof., 歯学部, 助教授 (50161245)

Co-Investigator(Kenkyū-buntansha) SHIN Sunok  School of Dentistry, Assistant Prof., 歯学部, 助手 (30226336)
KAMIYA Masako  School of Dentistry, Assistant Prof., 歯学部, 助手 (80181907)
YASHIRO Koji  School of Dentistry, Lecturer, 歯学部, 講師 (50182316)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 1994: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywordsparotid gland / exocytosis / secretory granule / apical plasma membrane / G-protein / cytoskeleton / in vitro exocytotic model / membrane phospholipid metabolite / 形質膜 / リン脂質分解酵素 / 基底膜側形質膜 / 1,2-ジアシル-sn-グリセロール / 低分子量GTP結合タンパク質 / rabタンパク質ファミリー
Research Abstract

It is very important to clarify the mechanism of secretion in the animal, in order to know how to communicate and regulate between both cells, and tissues. Exocytosis is the final stage of the stimulus-secretion coupling phenomenon which is linked to the membrane fusion between secretory granules and plasma membranes. In this study, the rat parotid gland cells were used as a model for secretory cells. A cell-free exocytosis system, reconstituted with isolated secretory granules and isolated apical plasma membranes was adopted to study the molecular mechanism of the final step of secretion. In this system, the indicators of the exocytosis were amylase release and the membrane fusion which was assayd by fluorescence dequenching. Consequently, the metabolites (free fatty acids, 1,2-diacyl-sn-glycerol, phosphatidic acid) from membrane phospholipids accelerated both the amylase release and membrane fusion. According to the analysis of the localization of phospholipid hydrolyzing enzymes, it … More was suggested that the metabolites were generated in the membrane region of particular organelles. These results show the existence of specific membrane sites and mechanisms for exocytosis. It was known that the activity of phospholipases A_2 and D was regulated by trimmer-type, and low molecular GTP-binding proteins (G-protein), respectively. The analysis found that the localization of both G-proteins in plasma membranes and in secretory granules was different. The question of how the metabolizing enzyme and G-protein regulate the exocytosis remains to be answered. It was also observed that the proteins of cytoskeleton were bound to plasma membranes, not to secretory granule membranes, and that they suppressed the amylase release and the membrane fusion in the cell-free system.
From the aforementioned, we can find the mechanism which decides the region where the exocytosis occurs, and parts of the regulatory mechanisms used in the acceleration and suppression of exocytosis at the molecular level. Less

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Koji Yashiro: "Compositions of fatty acid in 1,2-diacylglycerol,phosphatidic acid, triacylglycerol, and nonesterified fatty acid in rat submandibular and sulingual salivary glands" Japanese Journal of Oral Biology. 35. 205-212 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yutaka Yokota: "Regulation of phospholipid metabollism and its functional relations to sceretory exocytosis in rat salivary glands" Dentistry in Japan. 30. 56-62 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Sun-Ok Shin: "Characterication of microsomal long-chain acyl-CoA hydrolase activity in the rat submandibular gland" International Journal of Biochemistry. 26. 279-285 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Koji Yashiro: "Changes in microsomal lysophospholipid acyltransferase activity are correlated with rat parotid gland enlargement induced by chronic administration of isoproterenol" Journal of Biochemistry. 115. 1040-1046 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yasunaga Kameyama: "Localization of a low MrGTP-binding protein, rap1protein, in plasma membranes and secretory granule membranes of rat parotid gland" Life Sciences. 55. 213-219 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masako Mizuno-Kamiya: "The significance of membrane lipids in exocytosis:control of liposome-evoked amylase release from secretory granules isolated from the rat parotid gland" Journal of Biochemistry. (submitted).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Koji Yashiro: "Substrate specificity of microsomal l-acyl-sn^glycero-3-phosphoinositol acyltransferase in rat submandibular gland for polyunsaturated long-chain acyl-CoAs" Biochemica et Biophysica Acta. (submitted).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masako Mizuno-Kamiya: "ATP-mediated activation of Ca^<2+>-independent phospholipase A_2 in the secretory granular membranes from the rat parotid gland" Biochimica et Biophysica Acta. (submitted).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Koji Yashiro, Sun-Ok Shin, Atsuyoshi Himejima, Akira Mori, Masamichi Yosida, Fuyuakira Takatsu, Akira Shinkai, Yasunori Saito and Yutaka Yokota: "Compositions of fatty acid in 1,2-diacylglycerol, phosphatidic acid, triacylglycerol, and nonesterified fatty acid in rat submandibular and sublingual salivary glands" Jpn.J.Oral Biol.35. 205-212 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yutaka Yokota, Yasunaga Kameyama, Koji Yashiro, Masako Mizuno-Kamiya and Sun-Ok shin: "Regulation of phospholipid metabolism and its functional relations to secretory exocytosis in rat salivary glands" Dentistry in Japan. 30. 56-62 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] sun-Ok shin, Yasunaga Kameyama, Masamichi Yoshida, Fuyuakira Takatsu, Akira Shinkai, Hiroshi Inokuchi, Yasunori Saito and Yutaka Yokota: "Characterization of microsomal long-chain acyl-CoA hydrolase activity in the rat submandibular gland" Int.J.Biochem. 26. 279-285 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Koji Yashiro, Yasunaga Kameyama, Masako Mizuno-Kamiya, Sun-Ok shin, Akira Shinkai, Fuyuakira Takatsu and Yutaka Yokota: "Changes in microsomal lysophospholipid acyltransferase activity are correlated with rat parotid gland enlargement induced by chronic administration of isoproterenol" J.Biochem. 115. 1040-1046 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yasunaga Kameyama, Koh-ichi Nagata, Masako Mizuno-Kamiya, Yutaka Yokota, atsushi Fujita and Yoshinori Nozawa: "Localization of a low Mr GTP-binding protein, rapl protein, in plasma membranes and secretory granule membranes of rat parotid gland" Life Sci. 55. 213-219 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masako Mizuno-Kamiya, Yasunaga Kameyama, Koji Yashiro, Sun-Ok shin, Hiroshi Inokuchi and Atsushi Fujita: "The significance of membrane lipids in exocytosis : control of liposome-evoked amylase release from secretory granules isolated from the rat parotid gland" J.Biochem. submitted for publication.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Koji Yashiro, Yasunaga Kameyama, Masako Mizuno-Kamiya, Sun-Ok Shin and Atsushi Fujita: "Substrate specificity of microsomal 1-acyl-sn-glycero-3-phosphoinositol acyltransferase in rat submandibular gland for polyunsaturated long-chain acyl-CoAs" Biochim. Biophys. Acta. submitted for publication.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Masako Mizuno-Kamiya, Yasunaga Kameyama, Koji Yashiro, Sun-Ok Shin, Hiroshi Inokuchi and Atsushi Fujita: "ATP-mediated activation of Ca^<2+>-independent phospholipase A_2 in the secretory granular membranes from the rat parotid gland" Biochim. Biophys. Acta. submitted for publication.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Koji Yashiro: "Changes in microsomal lysophospholipid acyltransferase activity are correlated with rat parotid gland enlargement induced by chronic administration of isoproterenol" Journal of Biochemistry. 115. 1040-1046 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yasunaga Kameyama: "Localization of a low Mr GTP-binding protein,rapl protein,in plasma membranes and sccretory granule membranes of rat parotid gland" Life Sciences. 55. 213-219 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Akira Mori: "Analysis of membrane phospholipid and its fatty acid compositions in salivary glands of young female rats" Japanese Journal of Oral Biology. 35. 93-97 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Koji Yashiro: "Compositions of fatty acid in 1,2-diacylglycerol,phosphatidic acid,triacylglycerol,and nonesterified fatty acid in rat submandibular and sublingual salivary glands" Japanese Journal of Oral Biology. 35. 205-212 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yutaka Yokota: "Regulation of phospholipid metabolism and its functional relations to secretory exocytosis in rat salivary glands" Dentistry in Japan. (in press). (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Sun-OK Shin: "Characterization of microsomal long-chain acyl-CoA hydrolase activity in the rat submandibular gland" International Journal of Biochemistry. (in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Koji Yashiro: "Changes in microsomal lysophospholipid acyltransferase activity are correlated with gland enlargement induced by chronic administration of isoproterenol in rat parotid gland" Journal of Biochemistry. (in press). (1994)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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