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MOLECULAR MECHANISM FOR ALTERATION IN SIGNAL TRANSDUCTION BY OXIDATIVE STRESS -Isoform specific activation of C kinase by oxidized DAG and cell injury-

Research Project

Project/Area Number 10670216
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Experimental pathology
Research InstitutionTokai University

Principal Investigator

TAKEKOSHI Susumu  TOKAI UNIVERSITY, SCHOOL OF MEDICINE, ASSISTANT RESEARCHER, 医学部, 助手 (70216878)

Co-Investigator(Kenkyū-buntansha) WATANABE Keiichi  TOKAI UNIVERSITY, SCHOOL OF MEDICINE, PROFESSOR, 医学部, 教授 (00055865)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥2,200,000 (Direct Cost: ¥2,200,000)
KeywordsDIACYLGLYCEROL / PROTEIN KINASE C / LIPID PEROXIDATION / SIGNAL TRANSDUCTION / GLUTATHIONE PEROXIDASE / diacylglycerol / oxidative stress
Research Abstract

Protein kinase C (PKC) plays a crucial role a receptor-mediated signal transduction affecting diverse range of cellular responses such as cell proliferation, differentiation, and tumor promotion. PKC is a unique enzyme, which is activated by 1,2-diacylglycerol (DAG) produced from receptor-mediated hydrolysis of inositol phosphate. On the other hand, active oxygen species such as superoxide, hydrogen peroxide, and hydroxy radical have been suggested to play important roles in many pathological events such as inflammation, autoimmune diseases, ischemia-reperfusion injury, atherosclerosis and cardiovascular diseases, and carcinogenesis. This suggests a close relationship between the action of PKC and active oxygen species-induced dysfunction and cellular damage, however, this possibility has not been thoroughly examined. In the present study, we observed that 1-stearoyl-2-linoleoylglycerol hydroperoxide (SLG-OOH) and SLG Hydroxide (SLG-OH) stimulated the PKC activity isolated from rat bra … More in more efficiently than that of unoxidized SLG in the presences of phosphatidylserin and calcium ion. Then, the PKC activation effect of oxidized DAG has also been tried on 7 repsentative PKC isoforms, which were most pronouncedly expressed in rat brains, and alpha and delta isoforms were found to be intensely activated by the oxidized DAGs. Furthermore, we attempts to prove the neurodegenerative effects of DAG-OOH on cultured neurons overexpressing alpha and delta isoforms. As the cultured cells, neuronal cells established from 18-day rat fetus cerebral cortex (PN cells) were employed for the experiments. We used an adenovirus vector system in PN cells, which allows expression of PKC alpha and delta gene at a high level. These cultured neurons treated by streptolysin O to let the penetration of natural DAG and DAG-OOH into cells. Those treated cells were observed by 1) phase contrast microscopy, 2) cell viability, 3) electron microscopy, 4) immunoblot analysis of phoshorylated tau, 5) immunohistochemistry to localize phosphorylated tau. Within 6h of exposure to DAG-OOH, PN cells overexpressing PKC delta isoform, exhibited neuritic thinning and characteristic beading, while PN cells overexpressing alpha isoform did not shows such changes. The MEK inhibitor PD98059 prevented its neurodegeneration caused by DAG-OOH. Those lesions were observed routine EM which revealed the microtubule (MT) disassembly in the lesions. Immunoblot analysis showed that tau was partially phosphorylated on Thr 181, Ser 202 and Thr 205 in those DAG-OOH-treated cells. Phosphorylation of tau may have caused in MT disassembly and accumulated in cell body and beading lesions. Those findings indicate that the pronounced activation of PKC delta by DAG-OOH may specifically be responsible to elicit the neurodegeneration. Less

Report

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

    (18 results)

All Other

All Publications (18 results)

  • [Publications] Murakoshi M., Takekoshi T,, et al: "Immunolocalization of glutathione-peroxidase(GSH-PO) in the rat ventral prostate"J Toxicol Pathol. 10. 229-232 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuno A., Takekoshi S., et al: "Ultrastructural simultaneous identification of growth hormone and its messenger ribonucleic acid"Endocrin J. 45. 101-104 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuno A., Takekoshi S., et al: "An improved ultrastructural double-staining method for rat growth hormone and its mRNA using LR white rasin."Histochemical Journal. 30. 105-109 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Takagi T., Takekoshi S., et al: "Quercetin, a flavonol, promotes disassembly of microtubules in prostate cancer cells: Possible Mechanism of its antitumor activity"Acta Histochem. Cytochem. 31. 435-445 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nagata H., Takekoshi S., et al: "Antioxidative action of flavonids, Quercetin and Catechin mediated by the activation of glutathine peroxidase"Tokai J Exp Clin Med. 34. 1-11 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tajima A., Takekoshi S., et al: "distribution of adrenomedullin(AM), proadrenomedullin N-terminal 20 peptide, and AM mRNA in the rat gastric mucosa by immunocytochemistry and in situ hybridization"Histochem. Cell Biol. 112. 139-146 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Masanori Murakoshi, Rie Ikeda, Masashi Tagawa, Takaharu Nakayama, Susumu Takekoshi and Keiichi Watanabe: "Immunolocalization of glutathione-peroxidase(GSH-PO) in the rat ventral prostate"J Toxicol Pathol. 10. 229-232 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Akira Matsuno, Tadashi Nagashima, Susumu Takekoshi, Hirotoshi Utsunomiya, Naoko Sanno, R. Yoshiyuki Osamura, Keiichi Watanabe, Akira Tamura, Akira Teramoto: "Ultrastructural simultaneous identification of growth hormone and its messenger ribonucleic acid."Endocrin Journal. 45. 101-104 (198)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Akira Matsuno, Yoshitaka Ohsugi, Hirotoshi Utsunomiya, Susumu Takekoshi, Shigeru, Munakata, ,Koiichi Nagao, R. Yoshiyuki ,Osamura, Akira Tamura Tadashi Nagashima: "An improved ultrastructural double-staining method for rat growth hormone and its mRNA using LR white resin."Histochemical Journal. 30. 105-109 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tetsuo Takagi, Susumu Takekoshi, Takeshi Okabe, Hidetaka Nagata, Takao Honma, Keiichi Watanabe: "Quercetin, a flavonol, promotes disassembly of microtubules in postate carncer cells: Possible Mechanism of its antitumor activity."Acta Histochem. Cytochem.. 31. 435-445 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Hidetaka Nagata, Susum Takekoshi, Tetsuo Takegi, Takao Honma, Keiichi Watahabe: "Antioxidative action of flavonids, Quercetin and Catechin mediated by the activation of glutathione peroxidase,"Tokai J Exp Clin Med. 34. 1-11 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Akihiro Tajima, Yoshiyuki Osamura, Susumu Takekoshi, Yoshiko Itoh, Naoko Sanno, Tetsuya Mine, Toshiro Fujita: "Distribution of adrenomedullin (AM), proadrenomedullin N-terminal 20 peptide, and AM mRNA in the rat gastric mucosa by immunocytochemistry and in situ hybridization"Histochem Cell Biol. 112. 139-146 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nagata H., Takekoshi S., et al: "Nagata H., Takekoshi S., et al. Antioxidative action of flavonids, Quercetin and Catechin mediated by the activation of glutathione peroxidase"Tokai J Exp Clin Med. 34. 1-11 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Tajima A., Takekoshi S., et al: "Distribution of adrenomedullin (AM), proadrenomedullin N-terminal 20 peptide, and AM mRNA in the rat gastric mucosa by immunocytochemistry and in situ hybridization"Histochem Cell Biol. 112. 139-146 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Matsuno,A.,et al: "An improved ultrastructural double-staining method for rat growth hormone and its mRNA using LR white resin." Histochemical Journal. 30. 105-109 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Matsuno,A.,et al: "Ultrastructural simultaneous identification of growth hormone and its messenger ribonucleic acid." Endocrine Journal. 45. 101-104 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takagi,T.,et al: "Quercetin,a flavonol,promotes disassembly of microtubules in prostate cancer cells." Acta Histochem.Cytochem.(In press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Nagata,H.,et al: "Antioxidative action of flavonoids,quercetin and catechin,mediated by the activation of glutathione peroxidase." Tokai Experimental Clinical Medicine. (In press). (1999)

    • Related Report
      1998 Annual Research Report

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

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