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Analysis of cell polarity formation by gene transfection in organs in situ

Research Project

Project/Area Number 10470003
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General anatomy (including Histology/Embryology)
Research InstitutionGunma University School of Medicine

Principal Investigator

TAKATA Kuniaki  School of Medicine, Gunma University, Professor, 医学部, 教授 (20129290)

Co-Investigator(Kenkyū-buntansha) SUZUKI Takeshi  School of Medicine, Gunma University, Research Associate, 医学部, 助手 (00261868)
鈴木 雅一  群馬大学, 生体調節研究所, 助手 (60280913)
Project Period (FY) 1998 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥12,700,000 (Direct Cost: ¥12,700,000)
Fiscal Year 2001: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2000: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1998: ¥5,400,000 (Direct Cost: ¥5,400,000)
Keywordsliver / pancreas / electroporation / glucose transporters / immunohistochemistry / particle gun / immuno-electron microscopy / GLUT / 細胞膜 / GFP / 遺伝子導入 / 糖輸送体位 / 頂部細胞膜 / 基底側壁部細胞膜 / 臓器細胞 / 遺伝子直接導入法 / リポフェクション法 / ラット / 電気パルス
Research Abstract

In vivo electroporation and particle gun methods were used to introduce exogenous genes into tissue cells in situ in living animals. Constructs of green fluorescent proteins were tested to optimize the conditions of gene transfection. Glucose transporters (GLUT1, GLUT3, GLUT4, GLUT5, and SGLT1) were introduced into cells of liver and pancreas of living rats. The expressed gene products were visualized by immunofluorescence staining of tissue sections. By optimizing the conditions of transfection, glucose transporters were successfully expressed in highly differentiated cells such as hepatocytes and pancreatic acinar cells forming tissues in situ. GLUT1 was localized to basolateral membrane domains in both hepatocytes and pancreatic acinar cells. GLUT3 and GLUT5 were restricted to apical membrane domain in pancreatic acinar cells. They were found in both the apical and basolateral domains in hepatocytes. These observations suggest differential targeting mechanism in hepatocytes and acinar cells : i.e., apical targeting is carried out by direct pathway in acinar cells, whereas it is mediated by the indirect transcytotic pathway in hepatocytes. GLUT4 was retained in the cytoplasmic compartments in both cell types. Immuno-electron microscopy revealed that GLUT4 resides in the membranes of zymogne secretary granules in acinar cells, showing that GLUT4 is sorted to the regulated secretion pathway in these cells. In addition, in vivo electroporation and particle gun methods are simple and versatile methods to study the localization of exogenous gene products in cells and tissues in situ.

Report

(5 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (28 results)

All Other

All Publications (28 results)

  • [Publications] Shinoda Y: "Expression of sugar Trasporters by in vivo electroporation and particle gun method in the rat liver: localization to specific membrane domains"Acta Histochem Cytochem. 34. 15-24 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kanno T: "NMDA-receptor-dependent and -independent cytotoxic effect of D. discoideum differentiation-inducing factor on rat cortical neurons"Develop Growth Differ. 43. 709-716 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Takata K: "Glucose transporters-----Localization to specific membrane domains"電子顕微鏡. 36. 23-25 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 高田邦昭: "組織細胞化学2001(日本組織細胞化学会編)"学際企画. 8 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki T: "Direct gene transfer into rat liver cells by in vivo electroporation"FEBS Lett.. 425. 436-440 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sugawara-Yokoo M: "Presence of fructose transporter GLUT5 in the S3 proximal tubules in the rat"Kidney Int. 56. 1022-1028 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Takata K: "Sugar transporters in polarized epithelial cells"Acta Histochem Cytochem. 32. 53-58 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Matsuzaki T: "An anti-peptide antibody that recognized unexpected protein - A case report"Acta Histochem Cytochem. 33. 361-365 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki T: "Polarized distribution of Na+-dependent glucose cotransporter SGLT1 in epithelial cells"Acta Histochem Cytochem. 33. 159-162 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kanno T: "NMDA-receptor-dependent and -independent cytotoxic effect of D. discoideum differentiation-inducing factor on rat cortical neurons"Develop Growth Differ. 43. 709-716 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shinoda Y: "Expression of sugar transporters by in vivo electroporation and particle gun methods in the rat liver : Localization to specific membrane domains"Acta Histochem Cytochem. 34. 15-24 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki T: "The apical localization of SG1t1 glucose transporter is determined by the short amino acid sequence in its N-terminal domain"Eur J Cell Biol. 80. 765-774 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shinoda Y, Suzuki T, Sugawara-Yokoo M, Nagamatsu S, Kuwano H, Takata K.: "Expression of sugar transporters by in vivo electrotion and particle gun methods in the rat liver : Localization to specific membrane domains"Acta Histochem Cytochem. 34. 15-24 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kanno T.: "NMDA-receptor-dependent and -independent cytotoxic effect of D.discoideum differentlation-inducing factor on rat cortical neurons"Develop Growth Differ. 43. 709-716 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takata K.: "Glucose transporters-Localization to specific membrane domains"電子顕微鏡. 36. 23-25 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 高田邦昭: "組織細胞化学2001 (日本組織細胞化学会編)"学際企画. 8 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Omata W: "Actin filaments play a critical role in insulin-induced exocytotic recruitment but not in endocytosis of GLUT4 in isolated rat adipocytes."Biochem J. 346. 321-328 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sugawara-Yokoo M: "Differential localization of organic cation transporters rOCT1 and rOCT2 in the basolateral membrane of rat kidney tubules."Histochem Cell Biol. 114. 175-180 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Matsuzaki T: "An anti-peptide antibody that recognized unexpected protein-A case report."Acta Histochem Cytochem. 33. 361-365 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 高田邦昭: "遺伝子工学を用いた形質膜の機能解析"組織培養工学. 26. 170-171 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ogihara T: "Peptide transporter (PepT1) in the rat small intestine: ultrastructural localization and effect of starvation and administration of amino acids"Histochem J. 31. 169-174 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sugawara-Yokoo M: "Presence of fructose transporter GLUT5 in the S3 proximal tubules in the rat"Kidney Int. 56. 1022-1028 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takata K: "Sugar transporters in polarized epithelial cells"Acta Histochem Cytochem. 32. 53-58 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 高田邦明: "グルコース輸送体:その生理機能・病態と関連"Bio Clinica. 14(10). 893-898 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Suzuki T: "Direct gene transfer into rat liver cells by in vivo electroporation." FEBS Lett. 425. 436-440 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 高田邦昭: "糖輸送体と上皮細胞の極性." Mebio. 15(3). 129-131 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 高田邦昭: "上皮細胞における糖輸送機構." 解剖学雑誌. 73. 485-495 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takata K: "Sugar transporters in polarized epithelial cells." Acta Histochem Cytochem. (in press).

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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