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The role of transbilayer asymmetry of phospholipids in exocytosis

Research Project

Project/Area Number 14572037
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionNagoya City University

Principal Investigator

HIRASHIMA Naohide  Nagoya City University, Graduate School of Pharmaceutical Sciences, Associate Professor, 大学院・薬学研究科, 助教授 (10192296)

Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
Keywordsexocytosis / lipid scramblase / transbilayer asymmetry of phospholipids / flip-flop / mast cell / PC12 / cholesterol / flotillin / PC12
Research Abstract

In this study, we investigated the role of membrane components other than membrane protein such as phospholipids and cholesterol in exocytotic release.
1.The role of transbilayer asymmetric distribution of phospholipids in exocytosis
In order to disrupt the transbilayer asymmetry of phospholipids, lipid scramblase, which catalyze hi-directional movement of phospholipids between two membrane layers, was expressed in mast cells or PC12 cells. We found that exocytosis was significantly inhibited in these cells. This result suggests that transbilayer asymmetry of phospholipids plays an important role in exocytotic release.
2.The role of cholesterol in the plasma membrane in exocytotic process
The effects of cholesterol depletion from the plasma membrane with methyl-β-cyclodextrin (MβCD) on exocytotic processes were investigated in mast cells. Pretreatment with MβCD inhibited antigen-evoked exocytosis. The inhibition of exocytosis by MβCD was observed even under stimulation with phorbol ester a … More nd calcium ionophore. Therefore, MβCD affected membrane fusion between granule and the plasma membrane. Intracellular Ca^<2+> measurements revealed that MβCD inhibited Ca^<2+> influx from the extracellular medium through the store-operated calcium channel (SOC) without affecting Ca^<2+> from Ca^<2+> store. These results suggest that cholesterol depletion inhibits degranulation mainly by impairing SOC and exocytotic membrane fusion between granules and the plasma membrane.
3.Raft protein flotillin regulates IgE receptor mediated signaling
We investigated the role of flotillin, which is localized in lipid rafts, in the early exocytotic process. We obtained flotillin-1 knockdown RBL-2H3 cells. In these cells, we observed a significant decrease in Ca^<2+> tyrosine phosphorylation of IgE-R, and exocytosis. We also found that flotillin is associated with Lyn in lipid rafts. Antigen stimulation causes augmentation of flotillin binding to Lyn, resulting in increased activity of Lyn. These results suggest that flotillin-1 is an essential molecule in IgE-R mediated activation, regulating the kinase activity of Lyn. Less

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] Ari Noguchi: "Cationic cholesterol promotes gene transfection using the nuclear localization signal in protamine"Pharmaceut.Res.. 19. 933-938 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ari Noguchi: "Activation of receptor tyrosine kinases promotes gene transfection in rat neuronal PC12 cells by cationic liposomes"Neurosci.Lett.. 325. 29-32 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Naoto Kato: "Transbilayer asymmetry of phospholipids in the plasma membrane regulates exocytotic release in mast cells"Biochemistry. 41. 8068-8074 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Seiji Hasegawa: "Comparative Study of transfection Efficiency of Cationic Cholesterols Mediated by Liposomes-based Gene Delivery"Bioorg.Med.Chem.Lett.. 12. 1299-1302 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shoichiro Noguchi: "Remarkable induction of apoptosis in human cancer cells by a new cationic liposome complexed with a bc1-2 antisense oligonucleotide"J.Controlled Release. 88. 313-320 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shoichiro Noguchi: "Asialoganglioside Enhances the Efficiency of Gene Transfection Mediated by Cationic Liposomes with a Cationic Cholesterol Derivative"Biol.Pharm.Bull.. 26. 1306-1310 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Naoto Kato: "Cholesterol depletion inhibits store-operated calcium currents and exocytotic membrane fusion in RBL-2H3 cells"Biochemistry. 42. 11808-11814 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nobuyuki Ohyama: "The effects of ITIM-bearing FcRIIB on the nuclear shuttling of MAP kinase in RBL-2H3 cells"Immunol.Lett.. 90. 173-176 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yoshikazu Inoh: "MEL-A Dramatically increases gene transfection via membrane fusion"J.Controlled Release. 94. 423-431 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ari Noguchi: "Cationic cholesterol promotes gene transfection using the nuclear localization signal in protamine"Pharmaceut.Res.. 19. 933-938 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ari Noguchi: "Activation of receptor tyrosine kinases promotes gene transfection in rat neuronal PC12 cells by cationic liposomes"Neurosci.Lett.. 325. 29-32 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Naoto Kato: "Transbilayer asymmetry of phospholipids in the plasma membrane regulates exocytotic release in mast cells"Biochemistry. 41. 8068-8074 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Seiji Hasegawa: "Comparative study of Transfection Efficiency of Cationic Cholesterols Mediated by Liposomes-based Gene Delivery"Bioorg.Med.Chem.Lett.. 12. 1299-1302 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shoichiro Noguchi: "Remarkable induction of apoptosis in human cancer cells by a new cationic liposome complexed with a bcl-2 antisense oligonucleotide"J. Controlled Release. 88. 313-320 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shoichiro Noguchi: "Asialoganglioside Enhances the Efficiency of Gene Transfection Mediated by Cationic Liposomes with a Cationic Cholesterol Derivative"Biol.Pharm.Bull.. 26. 1306-1310 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Naoto Kato: "Cholesterol depletion inhibits store-operated calcium currents and exocytotic membrane fusion in RBL-2H3 cells"Biochemistry. 42. 11808-11814 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nobuyuki Ohayma: "The effects of ITIM-bearing FcRIIB on the nuclear shuttling of MAP kinase in RBL-2H3 cells"Immunol.Lett.. 90. 173-176 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yoshikazu Inoh: "MEL-A Dramatically increases gene transfection via membrane fusion"J. Controlled Release. 94. 423-431 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Naoto Kato: "Cholesterol depletion inhibits store-operated calcium currents and exocytotic membrane fusion in RBL-2H3 cells"Biochemistry, USA. 42. 11808-11814 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nobuyuki Ohyama: "The effects of ITIM-bearing FcRIIB on the nuclear shuttling of MAP kinase in RBL-2H3 cells"Immunol.Lett.. 90. 173-176 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Yoshikazu Inoh: "Biosurfactant MEL-A Dramatically increases gene transfection via membrane fusion"J.Controlled Release. 94. 423-431 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Naoto Kato: "Transbilayer asymmetry of phospholipids in the plasma membrane regulates exocytotic release in mast cells"Biochemistry, USA. 41. 8068-7084 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ari Noguchi: "Cationic cholesterol promotes gene transfection using the unclear localization signal in protamine"Pharm. Res.. 19. 933-938 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ari Noguchi: "Activation of receptor tyrosine kinases promotes gene transfection in rat neuronal PC12 cells by cationic liposomes"Neurosci. Lett.. 325. 29-32 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Seiji Hasegawa: "Comparative study of transfection efficiency of cationic cholesterols mediated by liposomes-based gene delivery"Bioorg. Med. Chem. Lett.. 12. 1299-1302 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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