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The molecular cloning in brain of El mice as refractory epileptic model.

Research Project

Project/Area Number 61570528
Research Category

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

Allocation TypeSingle-year Grants
Research Field Psychiatric science
Research InstitutionOsaka City University

Principal Investigator

KAWAKITA YUKIO  Osaka City University Medical School, 医学部, 教授 (80046820)

Co-Investigator(Kenkyū-buntansha) CHIKAME TAKASHI  Osaka City University Medical School, 医学部, 助手 (80163723)
ONISHI HIROSHI  Osaka City University Medical School, 医学部, 講師 (70094464)
大野 京介  大阪市立大学, 医学部, 助手 (90145780)
Project Period (FY) 1986 – 1987
Project Status Completed (Fiscal Year 1987)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1987: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1986: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsEl mouse / Epilepsy / Cloning / cloning / cDNA probe / hybridization / Elマウス脳 / poly(A)mRNA
Research Abstract

A single-stranded cDNA synthesized from El mouse brain poly(A^+)mRNA, was 350 to 400 nucleotide length and 38S of the sedimentation coefficient. The hybridized ratios of cDNA and poly(A^+)mRNAs from El(+), El(o) which did not convulse at all, and non-susceptible strain of ddY mice were 78, 75 and 55%, respectively, at Rot 10^5. The sequence complexity of nucleotides showed that there are three classes of abundant, intermediate and rare in mRNA, which values were 1.49 X 10^5, 4.64 X 10^6 and 1.87 X 10^7 nucleotides, respectively. These results imply that a significant difference in sequence complexity of abundant class was observed in genome between El and ddY mouse brains.
A cDNA library was prepared from El mouse brain poly(A^+)mRNA by the method of Okayama and Berg. Thousand colonies from this library were screened with excess of poly(A^+)mRNA in ddY or El mouse brains. In situe hybridization, the 500 positive clone showed a higher level of hybridization in El than ddY mouse brain. Plasmid DNA samples were digested with restriction endonucleases and the vector separated from insert fragments was carried out by gel electrophoresis. The Southern blot analysis revealed that the 200 selected brain cDNA clones ranged in abundance. Plasmid cDNA from each the 500 clones was nicked-translated and used to probe a Northern-blot analysis with poly(A^+)mRNA of El or ddY mouse brain. Two hundred clones showed a larger in hybridization intensity at El than ddY mouse brains. These results suggest that the difference of gene expression between El and ddY mice was induced during the process of posttranscription.

Report

(2 results)
  • 1987 Final Research Report Summary
  • 1986 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] 川北幸男ら: 神経化学. 25. 490-492 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 川北幸男ら: Jpn.J.Psychiat.Neurol.40. 361-365 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 川北幸男ら: Exp.Neurol.95. 167-177 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 川北幸男ら: Jpn.J.Psychiat.Neurol.41. 407-410 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 川北幸男ら: Jpn.J.Psychiat.Neurol.41. 588-589 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Y. Kawakita, et al.: "Sequence complexity of polyadenylate ribonucleic acid in El mouse brain" Jpn J. Psychiat. Neurol.40. 361-365 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Y. Kawakita, et al.: "Alteration of RNA synthesis in vitro in intact cerebral cortex nuclei induced by convulsions in seizure-susceptible El mice." Exp. Neurol.95. 167-177 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Y. Kawakita, et al.: "Someproperties of protein in synaptosomal fractions from El mouse cerebral cortices" Jpn J. Psychiat. Neurol.41. 407-410 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Y. Kawakita, et al.: "Change in synaptic proteins after seizures of El mice" Jpn J. Psychiat. Neurol.41. 588-589 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 川北幸男,他: Exptl.Neurol.83. 98-107 (1984)

    • Related Report
      1986 Annual Research Report
  • [Publications] 川北幸男,他: Folia Psychiat.Neurol.Jpn.39. 433-434 (1985)

    • Related Report
      1986 Annual Research Report
  • [Publications] 川北幸男,他: 神経化学. 24. 538-540 (1985)

    • Related Report
      1986 Annual Research Report
  • [Publications] 川北幸男,他: Jpn.J.Psychiat.Neurol.40. 357-361 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] 川北幸男,他: Exptl.Neurol.95. 167-177 (1987)

    • Related Report
      1986 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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