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The existence of NADPH oxidase in fish neutrophils

Research Project

Project/Area Number 08660236
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General fisheries
Research InstitutionMiyazaki University

Principal Investigator

IIDA Takaji  Miyazaki University, Associate Professor Faculty of Agriculture, 農学部, 助教授 (70159557)

Co-Investigator(Kenkyū-buntansha) ENDO Makoto  Miyazaki University, Associate Professor Faculty of Agriculture, 農学部, 助教授 (80128355)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1996: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsJapanese eel / neutrophil / NADPH oxidase / active oxygen / cytochrome b558 / respiratory burst / NADPH酸化酸素
Research Abstract

During respiratory burst, eel neutrophils consumed oxygen, and produced active oxigens. Superoxide production was equivalent to oxygen consumption, and hydrogen peroxide production was approximately half of oxygen consumption or superoxide production. This correlation is stoichiometrically identical to that of oxygen metabolism caused by NADPH oxidase during the respiratory burst in mammalian phagocytes, demonstrating the existence of the similar mechanism of oxygen radicals in fish neutrophils to mammals. The amounts of ocygen consumed and oxygen radicals produced in irritant-elicited neutrophils were significantly higher than those in unelicited neutrophils. This indicates that elicited neutrophils which possess high bactericidal activity were released from hematopoietic tissue to inactivate an exclude foreign irritants.
Cytochrome b558 is a membrance-bound component of NADPH oxidase present in mammalian phagocytes, and consisted of large and small subunits. Anti-Li antigocy against the synthetic peptide corresponding to the carboxyl terminal of human cytochrome b558 large subunit was produced. When western blotting was performed against Japanese ell neutrophil lysate, a broad band was specifically detected with the antibody at 80-110 kDa as well as human neutrophil smaple. Fluorescent antibody technique shows that the antibody recognized the epitope existed in eel neutrophils only after the cells were permeabilized with detergent. These results demonstrate tha the large subunit of cytochrome b558 exists in fish neutrophils and the caboxyl terminal is exposed to cytoplasmic sede, and suggest that NADPH oxidase is conserved in neutrophils across species.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] Takuya Itou: "Kinetics of oxygen metabolism during respiratory burst in Japanese eel neutrophils" Dev.Comp.Immunol.20(5). 323-330 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takuya Itou: "The importance of hydrogen peroxide in phagocytic bactericidal activity of Japanese eel neutrophils" Fish Pathol.32(2). 121-125 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takuya Itou: "Evidence for the existence of cytochrome b558 in fish neutrophils by polyclonal anti-peptide antibody" Dev.Comp.Immunol.(印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takuya Itou: "Kinetics of oxygen metabolism during respiratory burst in Japanese eel neutrophils" Dev.Comp.Immunol.20(5). 323-330 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takuya Itou: "The importance of hydrogen peroxide in phagocytic bactericidal activity of Japanese eel neutrophils" Fish Pathol.32(2). 121-125 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takuya Itou: "Evidence for the existence of cytochrome b558 in fish neutrophils by polyclonal anti-peptide antibody" Dev.Comp.Immunol.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takuya Itou: "Evidence for the existence of cytochrome b558 in fish neutrophils by polyclonal anti-peptide antibody" Dev.Comp.Immunol.,. (印刷中).

    • Related Report
      1997 Annual Research Report
  • [Publications] Takuya Itou: "The importance of hyfrogen peroxide in phagocytic bactericidal activity by Japanese eel neutrophils (印刷中)" Fish Pathol.32(2). (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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