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Mechanisms of recognition of non-self and initiation of defense reactions in insects

Research Project

Project/Area Number 02454019
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 動物発生・生理学
Research InstitutionHokkaido University

Principal Investigator

ASHIDA Masaaki  The Institute of Low Temperature Science ; Professor, 低温科学研究所, 教授 (50012422)

Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1990: ¥4,800,000 (Direct Cost: ¥4,800,000)
KeywordsINSECT / HEMOLYMPH / PHENOLOXIDASE / CUTICLE / CECROPIN / DEFENSE / PROTEASE / PEPTIDOGLYCAN / 異物認識 / フェノ-ル酸化酵素 / ペプチドグリカン / カビ / バクテリア / フェノ-ル酸化酵素前駆体 / 蚊 / ゴキブリ / バッタ
Research Abstract

The ultimate end of this research project is to study the mechanism by which foreign materials such as bacteria, fungi and parasites invading insects are recognized as non-self and how the recognition mechanism of non-self is linked to the initiation of defense reactions of insects. To reach the end the present research was carried out using two insect tissues, hemolymph and integument of the silkworm, Bombyx mori.
The prophenoloxidase cascade (ProPO cascade) is present in hemolymph. The cascade is triggered by minute amounts of beta-1,3-glucan (beta G) or peptidoglycan (PG). We have shown previously that PG and betaG trigger the cascade when they bind specifically to peptidoglycan recognition protein (PGRP) and beta-1,3-glucan recognition protein (beta GRP), respectively. Although it is speculated that the recognition proteins may play a central role in the recognition of bacteria or fungi as non-self in insect hemocoel, our knowledge of the chemical structures and the mode of action o … More f the recognition proteins is very limited. We investigated the localization of the recognition proteins in hemocoel by immunochemical and immunocytochemical methods and put forward a working hypothesis on the role of the two recognition proteins in the process of recognition of bacteria or fungi as non-self in hemocoel.
The components of proPO cascade include more than two serine enzyme zymogens. Experimental results so har obtained suggest that active forms of the zymogens or fragments derived from the zymogens during their activation modulate phagocytotic activity and hemokinetic activity of hemocytes. To prove this possibility unambiguously we are currently trying to purify all proPO cascade components. We recently have succeeded in purifying proBAEEase and its active form to homogeneity. It is expected that the active enzyme will be studied whether it has the effect on the phagocytotic or hemokinetic activity of hemocytes.
Cuticle of insect integument is often considered to be a protective armor and chemically inert (dead) part of the body. Using axenically raised larvae of silkworm, Bombyx mori we could demonstrate that the cuticle transmit a signal to epidermal cells of integument to direct them to synthesize antibacterial peptide (cecropin) when the cuticle has bacterial cell wall component (lipopolysaccharide or peptidoglycan) on its abraded part. The antibacterial peptide is secreted to cuticle. The presence of bacterial cell wall component on abraded cuticle was shown to cause the synthesis of cecropin mRNA in fat body, but in this case no cecropin activity was detected in fat body and hemolymph. Seemingly this mRNA synthesis without immediate translation of the message indicate that cuticular abrasion with bacterial cell wall component make the the entire insect body alert to bacterial infection. Currently we are investigating the nature of signal sent to epidermal cells and fat body from the abraded part of cuticle with bacterial cell wall component. Less

Report

(4 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • 1990 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Ashida,M.,Kinoshita,K.and Brey.P.T.: "Studies on prophenoloxidase activation in the mosquito Aedes aegypti" L.Eur.J.Biochem.188. 507-515 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ashida,M.: "The prophenoloxidase cascade in insect immunity." Res.lmmunol.141. 908-910 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Brey P.T.,Stoltz,D.B.,Cook,D.I.,Ashida,M.: "Use of nitrocellulose membrane to activate and measure insect prophenoloxidase." Anal.Biochem.194. 359-364 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ochiai,M.,Niki,T.,Ashida,M.: "Immunocytochemical localization of β-1,3-glucan recognition protein in the silkworm,Bombyx mori." Cell & Tissue Res.268. 431-437 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Bery,P.T.,Lee W-J.,Yamakawa,M.,Koizumi,Y.,Perrot,S.,Frangoi,M.and Ashida,M.: "Role of integument in insect immunity:Epicuticular abraision and induction of cecropin synthesis in cuticular epithelial cell." Proc.Natn.Acad.Sci USA.(1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ashida,M.,Sasaki,T.: "A target protease of serpins in insect hemolymph is an endogenous protease of prophenoloxidase cascade." Insect Biochem.and Molec.Biol. (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 芦田 正明: "昆虫の血液" 遺伝. 45. 40-45 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 芦田 正明,土屋 正和: "昆虫の特異な生体防御反応系と利用技術" ブレインテクノニュース. 31. 1-5 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 芦田 正明: "昆虫の特異な生体防御反応系と利用技術(フェノール酸化酵素カスケード)ー昆虫の生体防御反応における役割および医療機器の汚染検査薬としての可能性ー" 蚕系・昆虫農業技術研究所研究資料. 11. 19-24 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ashida,M.,Yamazaki,H.I.: ""Biochemistry of the phenoloxidase system in insects:with special reference to its activation"in Molting and Metamorphosis (Ohnish,E.and Ishizaki,K.Eds.)" Japan Sci Soc.Press,Tokyo/Springer Verlag,Berlin, PP239-265 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 芦田 正明: "学会出版センター(名取俊二,野本亀久雄,古田恵美子,松村繁 編)" "フェノール酸化酵素カスケードー昆虫の生体防御反応における役割", PP111-126 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ashida, M., Kinoshita, K. and Brey, P.T.: "Stidies on prophenoloxidase activation in the mosquito Aedes aegypti" Eur. J. Biochem. 188. 507-515 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ashida, M.: "The prophenoloxidase cascade in insect immunity." Research in Immunol. 141. 908-910 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Brey, P.T., Stoltz, D.B., Cook, D.I. and Ashida, M.: "Use of nitrocellulose membrane to activate and measure insect phenoloxidase." Anal. Biochem.194. 359-364 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ochiai, M., Niki, T. and Ashida, M.: "Immunocytochemical localization of beta-1.3-glucan recognition protein in the silkworm, Bombyx mori." Cell & Tissue Res. 268. 431-437 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Brey, P.T., Lee, U-J., Yamakawa, M., Koizumi, Y., Perrot, S., Francoi, M. and Ashida, M.: "Role of integument in insect immunity: Epicuticular abrasion and induction of cecropin synthesis in cuticular epithelial cells." Proc. Natn. Acad. Sci. U.S.A.(1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ashida, M. and Sasaki, T.: "A target protease of serpins in insect hemolymph is an endogenous protease of prophenoloxidase cascade." Insect Biochem. & Molec. Biol.(1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ashida, M. and Yamazaki, H.I.: "Biochemistry of the phenoloxidase system in insects with special reference to its activation" In Molting and Metamorphosis (Ohnishi, E. and Ishizaki, K. eds.). Japan Sci.Soc. Press, Tokyo/Springer Verlag, Berlin. 239-265 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ochiai,M.and Ashida,M.: "lmmunocytochemical localization of β-1,3-glucan recognition protein in the silkworm,Bombyx mori." Cell & Tissue Research. 268. 431-437 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Brey.P.T.et al.: "Role of The integument in insect immunity:Epicuticrlar abrasion and induction of cecuopin synthesis in cuticular epithirial cells." Proc.Nath.Acad.Sci.U.S.A.(1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Asida,N.and Sasaki,T.: "A target protease of sepins in insect hemolymoh is an endogenous protease of proohenoloxidase cascade." Insect Biochem.and Molec.Biol.(1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] 芦田 正明,土谷 正和: "昆虫の特異な生体防御反応系と利用技術" ブレインテクノニュース. 31. 1-5 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] 名取 俊二 他: "無脊椎動物の生体防御" 学会出版センター, 287 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] P.T.Brey,D.B.Stoltz,D.I.Cook&M.Ashida: "Use of nitrocellulose membrane to activate and measure insect phenoloxidase" Anal.Biochem.194. 359-364 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] M.Ochiai,T.Niki&M.Ashida: "Immunoーcytochemical localization of βー1,3ーglucan recognition protein in the silkworm,Bombyx mori." Cell and Tissue Res.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Ashida,M: "The prophenoloxidase cascade in insect immunity" Res.Immunol.141. 908-910 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Brey,P.T.et al: "Use of nitrocellulose membrane to activate and measure insect phenoloxidase." Anal.Biochem.(1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Yoshida,H.et al: "Purification of peptidoglycan recognition protein of the prophenoloxidase cascade in hemolymph of the silkworm,<Bombyx>___ー <Mori>___ー." J.Biol.Chem.

    • Related Report
      1990 Annual Research Report
  • [Publications] Kinoshita,K.et al: "Immunoーlocalization of peptidoglycan recognition protein in hemolymph of the silkworm,<Bombyx>___ー <Mori>___ー." Tissue and Cell Res.

    • Related Report
      1990 Annual Research Report
  • [Publications] Ashida,M.and Yamazaki,H.I.: "Biochemistry of the Phenoloxidase system in Insects:with Special Referense to its Activation." Molting and Metamorphosis (Ed.by E.Ohnish and H.Ishizaki) Japan Sci,Soc.Press/SPringerーVerlag Berlin.239-265 (1990)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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