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Analysis of regulatory mechanism for endoplasmic reticulum proliferation through DNA-binding factors

Research Project

Project/Area Number 09470005
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General anatomy (including Histology/Embryology)
Research InstitutionKansai Medical University

Principal Investigator

KANAMURA Shinsuke  Kansai Medical University, Department of Anatomy, Professor, 医学部, 教授 (50027091)

Co-Investigator(Kenkyū-buntansha) TAKAMORI Yasuharu  Kansai Medical University, Department of Anatomy, Instructor, 医学部, 助手 (50309233)
NIHIRA Mika  Kansai Medical University, Department of Anatomy, Instructor, 医学部, 助手 (20274194)
WATANABE Jun  Kansai Medical University, Department of Anatomy, Associate Professor, 医学部, 助教授 (40148557)
MONDO Hiroko  Kansai Medical University, Department of Anatomy, Instructor, 医学部, 助手 (30309249)
近藤 千雅  関西医科大学, 医学部, 助手 (20298863)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥12,300,000 (Direct Cost: ¥12,300,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1998: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥8,600,000 (Direct Cost: ¥8,600,000)
KeywordsCYP2B1 / 2 / acyltransferase / endoplasmic reticulum / phenobarbital / mRNA / in situ hybridization / luminessence / CYP1A1 / CYP2B2 / 小胞体膜 / Phenobarbital / luminessence / Cytochrome P450 2B2 / 転写調節因子 / 生物発光 / 光子計測 / 膜合成 / cytochrome P-450 / 肝
Research Abstract

In the present study, we analyzed regulatory mechanism for endoplasmic reticulum proliferation through DNA-binding factors by a combination of morphological methods, such as highly sensitive cytochemical methods and stereological analyses, with molecular biological techniques in rat hepatocytes.
(1) Proliferation of the endoplasmic reticulum (ER) membrane in hepatocytes is independent of the induction of membrane-bound proteins. However, a drug, phenobarbital (PB), acts as a trigger for both proliferation of ER and induction of PB-inducible membrane-bound proteins, CYP2B1/2.
(2) Proliferation of ER by PB administration is not caused by suppression of membrane degradation but caused by promotion of biosynthesis of the ER membrane. PB induces glycerophosphate acyltransferase (GAT), a key enzyme for biomembrane synthesis.
(3) Interaction of PB reguratory element (PBRE) with proximal promoter in CYP2B1/2 genes is necessary to activate CYP2B1/2 gene transcription.
(4) We have developed a) in situ Southwestern hybridization technique, b) quantitative immunohistochemical methods to measure each CYP content in hepatocyte cytoplasm and c) a highly sensitive detection method with chemiluminessence and photon counting. By using these techniques, we have also revealed the regulatory mechanism for transcription activation of CYP1A1/2 genes through a DNA-binding factor (heterodimer of AhR and Arnt) in rat hepatocytes.

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (41 results)

All Other

All Publications (41 results)

  • [Publications] Tanaka, T.: "Quantitative analysis of endoplasmic reticulum and cytochrome P-450 in hepatocytes from rats injected with methylcholanthrene"European J. Cell Biol.. 74. 20-30 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakamura, M.: "Immunohistochemical localization of type II and type I collagens in articular cartilage of the femoral head of dexamethasone-treated rats"Histochem. J.. 29. 645-654 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe, J.: "Examination of relationship between immunostaining intensity and antigen amount with an antigen-immobilized filter model system and sections"Acta Histochem. Cytochem.. 30. 551-556 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe, J.: "Determination of accuracy of quantitative immunohistochemical results with an antigen-immobolized filter model system"Acta Anat. Nippon.. 73. 25-31 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Shibano, K.: "Culture of stromal cells derived from medullary cavity of human long bone in the presence of 1, 25-dihydroxyvitamin D_3, recombinant human bone morphogenetic protein-2, or ipriflavone"Bone. 22. 251-258 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Asaka, Y.: "Localization of xenobiotic-responsive element binding protein in rat hepatocyte nuclei after methelcholanthrene administration as revealed by in situ southwestern hybridization"J. Histochem. Cytochem.. 46. 825-832

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Mino, K.: "Effect of 3-methylcholanthrene administration on expression of cytochrome P-450 isoforms induced by phenobarbital in rat hepatocytes"J. Histochem. Cytochem.. 46. 1151-1160 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe, J.: "Changes in the density of cytochrome P-450 2B1/2 molecules in the membrane of endoplasmic reticulum in perivenular, midzonal and periportal hepatocytes of rats after administration of phenobarbital"Acta Histochem. Cytochem.. 32. 321-326 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe, J.: "Amplification of immunostaining intensity by the peroxidase-antiperoxidase, avidin-biotin-peroxidase complex or catalyzed signal amplification method gives erroneous antigen content in sections"Acta Histochem. Cytochem.. 32. 407-413 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kanamura, S.: "Cell Biology of cytochrome P-450 in the liver"Int. Rev. Cytol.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe, J.: "Effect of phenobarbital on intralobular expression of CYP2B1/2 in livers of rats. Difference in the expression between signal and repetitive administrations"Biochem. Pharmacol.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takeda, T.: "Rifampicin suppresses hepatic CYP2E1 expression and minimizes DNA injury caused by carbon tetrachloride in perivenular hapatocytes of mice"Alchol. Clin. Exp. Res.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 金村秦輔: "組織細胞化学1997 酵素組織化学の応用-光顕レベルを中心として-"学際企画. 239 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 金村秦輔: "組織細胞化学1998 組織化学反応量的解析の最新の進歩"学際企画. 209 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tanaka T.: "Quantitative analysis of endoplasmic reticulum and cytochrome P-450 in hepatocytes from rats injected with methlylcholanthrene."Eur. J. Cell. Biol.. 74. 20-30 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakamura M.: "Immunohistochemical localization of type II and type I collagens in articular cartilage of the femoral head of dexamethasone-treated rats."Histochem. J.. 29. 645-654 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe J.: "Examination of relationship between immunostaining intensity and antigen amount with an antigen-immobilized filter model system and sections"Acta Histochem.Cytochem.. 30. 551-556 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe J.: "Determination of accuracy of quantitative immunohistochemical results with an antigen-immobilized filter model system"Acta Anat. Nippon. 73. 25-31 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Shibano K.: "Culture of stromal cells derived from medullary cavity of human long bone in the presence of 1,25-dihydroxyvitamin D3, recombinant human bone morphogenetic protein-2 or ipriflavone."Bone. 22. 251-258 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Asaka Y.: "Localization of xenobiotic responsive element-binding protein in rat hepatocyte nuclei after methylcholanthrene administration as revealed by the situ Southwestern hybridization"J. Histochem. Cytochem.. 46. 825-32 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Mino K.: "Effect of 3-methlycholanthrene administration on expression of cytochrome P-450 isoforms induced by Phenobarbital in rat hepatocytes"J. Histochem. Cytochem. 46. 1151-1160 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe J.: "Changes in the density of cytochrome P-450 2B1/2 molecules in the membrane of endoplasmic reticulum in perivenular, midzonal and periportal hepatocytes of rats after administration of Phenobarbital."Acta Histochem. Cytochem.. 32. 321-326 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe J.: "Amplification of immunostaining intensity by the peroxidase-antiperoxidase, avidin-biotin-peroxidase complex or catalyzed signal amplification method gives erroneous antigen content in sections."Acta Histochem. Cytochem.. 32. 407-413 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kanamura S.: "Cell biology of cytochrome P-450 in the liver"Int. Rev. Cytol.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe J.: "Effect of phenobarbital on intralobular expression of CYP2B1/2 in livers of rats. Difference in the expression between single and repetitive administrations."Biochem. Pharmacol.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takeda T.: "Rifampicin suppresses hepatic CYP2E1 expression and minimizes DNA injury caused by carbon tetrachloride in perivenular hepatocytes of mice."Alchol. Clin. Exp. Res.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Watanabe, J.: "Changes in the density of cytochrome P-450 2B1/2 molecules in the membrane of endoplasmic reticulum in perivenular, midzonal and periportal hepatocytes of rats after administration of phenobarbital."Acta Histochem. Cytochem.. 32. 321-326 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Watanabe, J.: "Amplification of immunostaining intensity by the peroxidase-antiperoxidase,avidin-biotin-peroxidase complex or catalyzed signal amplification method gives erroneous antigen content in sections."Acta Histochem. Cytochem.. 32. 407-413 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kanamura, S.: "Cell Biology of cytochrome P-450 in the liver."Int. Rev. Cytol.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Watanabe, J.: "Effect of phenobarbital on intralobular expression of CYP2B1/2 in livers of rats. Difference in the expression between signal and repetitive administrations."Biochem. Pharmacol.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Takeda, T.: "Rifampicin suppresses hepatic CYP2E1 expression and minimizes DNA injury caused by carbon tetrachloride in perivenular hepatocytes of mice."Alchol. Clin. Exp. Res.. (in press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Mino K: "Effect of 3-methylchoalnthrene administration on expression of cytochrome P-450 isoforms induced by phenobarbital in rat hepatocytes" J Histochem Cytochem. 46. 1151-1160 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Watanabe J: "Determination of accuracy of quantiative immunohistochemical results with an antigen-immobilized filter model system." Acta Anat Nippon. 73. 25-31 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Asaka Y: "Localization of xenobiotic responsive element-binding protein in rat hepatocyte nuclei after methylcholanthrene administration as revealed by in situ Southwestern hybridization." J Histochem Cytochem. 46. 825-832 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shibano K: "Culture of stromal cells derived from medullary cavity of human long bone in the presence of 1,25-dihydroxyvitamin D3,recombinant human bone morphogenetic protein-2 or ipriflavone." Bone. 22. 251-258 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 田中富弥: "Quantitative analysis of enduplasimc reticulum and cytochrome P-450 in hepatocytes from rats injected with methylcholanthrene" European Jourral of Cell Bialogy. 74. 20-30 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 中村 誠也: "Immunohistochemical localization of type II and type I collagens in articular cartilage of the femoral head of dexamethasone-treated rats" Histochemical Journal. 29. 645-654 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 柴野 恵介: "Culture of stromol cells derived from medullary cavity of human long bcne in the presence of 1.25-d hydroxyuitamin D_3,recombinant human bcne morphogenic protein-2 or ipriflavone" Bone. 22. (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 渡辺 淳: "Examination of velationship between immunostaining intensity and antigen amount with an antigen-immobilized filter model system and sections" Acta Histochemica E+ Cytochemica. 30・506. (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 渡辺 淳: "Determinalion of accuracy of guantitative immunohistochemical results with an antigen-immobilized filter model system" Acta Anatomica Nipponica. 73・1. (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 金村 秦輔: "細織細胞化学 1997" 学際企画, 239 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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