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Studies on outgrowth of pseudopodial cables and spicule rod formation in cultured derived from micromeres of sea urchin embryos.

Research Project

Project/Area Number 03454026
Research Category

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

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

Principal Investigator

YASUMASU Ikuo  Waseda University, School of Education, Department of Biology, Professor, 教育学部, 教授 (10063421)

Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1993: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1992: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1991: ¥2,700,000 (Direct Cost: ¥2,700,000)
KeywordsSea Urchin Embryos / Micromere / Growth of Pseudopodial Cable / Spicule Rod Formation / Protein Kinase / Insulin / RNA Synthesis / Protein Phosphorylation / プロティンチロシンキナーゼ / GTP結合タンパク質 / 百日咳毒素 / プロテインチロシンキナ-ゼ
Research Abstract

Micromere-derived cells of sea urchin embryos are known to produce pseudopodial cables and spicule rods during culture with horse serum or blastocoelic fluid isolated from sea urchin embryos. In the present study, insulin was found to induce the cable growth without spicule rod formation. Insulin binding protein was found by SDS-PAGE of insulin treated cells. Insulin bound to the cells with Kd of about 0.5 pM and bound insulin was replaced by unknown compounds in horse serum and blastocoelic fluid. This indicates that horse serum and blastocoelic fluid contain insulin-like compounds. Insulin binding in the cells increased in 6 hr period due to translocation of the receptor protein from microsome to plasma membrane and was maintained high up to 40 hr of culture in the absence of horse serum or insulin. In the cells with high insulin binding capacity, insulin, as well as horse serum, augmentation of RNA synthesis occurred following activation of protein phosphorylation and finally resulted in the cable growth. Inhibition of protein phosphorylation reduced the rate of RNA synthesis and inhibition of RNA synthesis by actinomycin D after insulin-induced augmentation of protein phosphorylation resulted in blockage of pseudopodial cable grwoth. In the cells kept with horse serum, the rates of protein phosphorylation and RNA synthesis remained high even after the initiation of cable growth. Actinomycin D introduced after the cable growth initiation allowed the cells treated with horse serum or insulin to cause pseudopodial cables but blocked the spicule formation in horse serum-treated cells. Horse serum and blastocoelic fluid seems to contain signalling compounds other than insulin.

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] S.Kuno: "The changes in insulin-binding capacity in plasma membrane fraction during culture in vitro of the cells derived from micromeres of 16-cell-stage sea urchin embryos." Development,Growth & Differentiation. 36 (in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] S.Kuno: "Insulin-induced outgrowth of pseudopodial cables in cultured micromere-derived cells isolated from sea urchin embryos at the 16cell stage,with special reference to the insulin-receptors." Development,Growth & Differentiation. 36 (in press). (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] S.Kuno: "Several cell responces to insulin in cultured cells derived from micromeres,isolated from sea urchin embryos at the 16cell stage." Development,Growth & Differentiation. (submitted). (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kuno, Shin-ichi: "The changes in insulin-binding capacity in plasma membrane fraction during culture of the cells derived from micromeres of the 16 cell stage sea urchin embryos." Development, Growth & Differentiation. Vol.36, (in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kuno, Shin-ichi: "Insulin-induced outgrowth of pseudopodial cables in cultured micromerederived cells isolated from sea urchin at the 16 cell stage, with special reference to the insulin-receptors." Development, Growth & Differentiation. Vol.36, (in press). (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kuno, Shin-ichi: "Several cell responces to insulin in cultured cells derived from micromeres, isolated from sea urchin embryos at the 16 cell stage." Development, Growth & Differentiation. (Submitted).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kuno,Shin-ichi: "The changes in insulin-binding capacit in plasma membrane fraction during culture of the cells derived from micromeres of the 16 cell Stage sea urchin embryos" Development,Growth and Differentiation. 36(in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kuno,Shin-ichi: "Insulin-induced outgrowth of pseudopodial cables in cultured micromerederived cells isclated from Sea urchin at the 16 cell stage,with special referenc to the insulin-receptors" Development,Growth and Differentiation. 36(in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kamata,Yasuyuki: "Identification of GTP-biuding by ADP-ribosylation in the presence of Cholera tuxin,Pertursis toxin and botulimm tuxin in Plasma membrane isolated from eggs and lmbryos of sea urchin." Development,Growth and Differentiation. 34. 211-222 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Kamata: "Identification of GTP-binding Protein by ADP-ribosylation in The Prescrce of Cholere toxin,Pertussis toxin and Botulinum toxin D in Plasma membrene isolated from eggs and embryos of sea urchin" Development,Growth & Differentiation. 34. 211-222 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Kuno: "Inhibitory effects of actinowycin D on the growth of Pseudopodia cables and the formation of spiculerod in micromere-derived cells isolated from Hre 32-Cell stage cmbryos of sea urchin" Development,Growth & Differentiation.

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Kuno: "Insulin-induced growth of psendopodial cables in micromere-derived cells isolated from sea urchin embryos" Development,Growth & Differentiation.

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Kamata: "Identification of GTP-binding protein by ADP-ribosylation in the presence of cholera toxin,pertursis toxin and botulinum toxin in Plasma membrane isolated from eggs and embryos of sea urchin." Development,Growth & Differentiation. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Kuno: "Effect of actinomycin D on the growth of pseudopodial Cables and spicule formation in culture cells derival from micromeres isolated from sea urchin embryos,with special reference to cnanges in the activities of protein kinase c and H^+,K^+-ATPase." Development,Growth & Differentiation.

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Kuno: "Does phosphory lation of tyrosin residue in protein induced by insulin initiate growth of pesudopodia in cultured cells derived from micromeres isolated from sea urchin embryos."

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Kuno: "Possible participation of G-proteins in initiation of spicule formatiod in Culturen cells derived from isolated micromeres of sea urchin embryos."

    • Related Report
      1991 Annual Research Report
  • [Publications] K.Mitsunaga-Nakatsubo: "Stimulating effect of ruthenium red on pseudopodial cable Growth in micromeres derived from isolated micromeres of sea urchin embryos."

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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