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Development Diagnostic Method of Brain Disorders Using DNZ Probe and Monoclonal Antribody.

Research Project

Project/Area Number 60870032
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field Neurology
Research InstitutionKEIO UNIVERSITY SCHOOL OF MEDICINE

Principal Investigator

TSUKADA Yasuzo  KEIO UNIV. SCHOOL OF MED., PROFESSOR, 医学部, 教授 (00050956)

Co-Investigator(Kenkyū-buntansha) TOYA Shigeo  keio univ. school of med., rpfessor, 医学部, 教授 (40051205)
FUJISHIRO Masatoshi  KEIO UNIV.SCHOOL OF MED., INSTRUCTOR, 医学部, 助手 (40173421)
TAKAMATSU Ken  KEIO UNIV. SCHOOL OF MED., ASSISTANT PROFESSOR, 医学部, 講師 (90154898)
KOHSAKA Shin-ichi  KEIO UNIV. SCHOOL OF MED., ASSOCIATE PROFESSOR, 医学部, 助教授 (50112686)
MIKOSHIBA Katsuhiko  OOSAKA UNIV. PROTEIN INST., PROFESSOR, 蛋白質研究所, 教授 (30051840)
長池 一博  三菱化成総合研究所, 主任研究員
Project Period (FY) 1985 – 1987
Project Status Completed (Fiscal Year 1987)
Budget Amount *help
¥11,400,000 (Direct Cost: ¥11,400,000)
Fiscal Year 1987: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1986: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1985: ¥7,500,000 (Direct Cost: ¥7,500,000)
KeywordsMonoclonal antibody / DNA probe / Fluoroimmuno assay / Immunohistochemistry / Experimental model of brain injury / Experimental model of brain tumor / モノクローン抗体 / CNP / ミエリン / PLP / MBP / 遺伝子診断
Research Abstract

Diagnosis with monocional antibodies.
i) Production of the monoclonal antibody.
Monoclonal antibody (TFK-2) against CNP which is a biochemical marker of myelin sheath was produced using SP-2 as a parental cell. TFK-2 recognized CNP molecules in CNS but did not cross-react with that in PNS.
ii) Method of the Fluoroimmunoassay for CNP in CSF.
Sanswich method using two different type of antibodies to CNP were used in the Fluoro- immunoassay. And add another type of antibody. After washing the plate. Eropium labeled seceond antibody were added. And the fluorescence activity trapped in the platge were measured. Coreration between the degree of the brain damage and the CNP level in CSF were obsereved.
iii) Immunohistochemical studies on the myelin deficient mutant (mld) mice were performed using monoclonal antibodies against myelin specific proteins. In mld mice, myelin sheath in CNS were stained in some places, and it was suggested that MBP was expressed efficiently in mld mice.
iv) The participation of the oncogenes in neoplastic process of the brain tumors were studied immunohistochemically using monoclonal antibodies against oncogene products. In some human malignant astrocytoma, src gene product or-EGF-R were expressed.
2) Diagnosis with DNA probes.
i) Myelin deficient mutant mice (mld, jimpy).
in mld mice, abnormal duplication of the MBP gene was obsreved and it was the cause of inefficient transcription of the MBP gene caused hypomyelination. In jimpy mice, there might be miner mutation in the intron of the PLP gene.
ii) Brain tumors.
In some human malignant astrocytoma, gene amplification and over expression of EGF-R might be implicated in the neoplastic process. ErbB-2 might be associated with the tumorigenesis of malignant astrocytoma and malignant neurinoma induced by ENU in rats.

Report

(3 results)
  • 1987 Final Research Report Summary
  • 1986 Annual Research Report
  • 1985 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Kohsaka S.; Mita K.; Tsukada Y.; et al.: Journal of Neurchemistry. 44. 406-410 (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kimura M.; Mikoshiba K.; Tsukada Y.; et al.: Journal of Neurochemistry. 44. 692-696 (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Fujishiro M.; Kohsaka S. Tsukada Y.; et al.: Journal of Neurochemistry. 47. 191-195 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Inoue Y.; Mikoshida K.; Tsukada Y.; et al.: Developmental Brain Research. 26. 239-247 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takamiya Y.; Kohasaka S.; Toya S.; et al.: Brain Rsearch. 383. 305-309 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kohsaka S.; Yoshida K.; Inoue Y.; et al.: Brain rsearch. 372. 137-142 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kohsaka S.,Mita K., Tsukada Y., et al.: "Impaired development of rat cerebellum induced by neonatal injection of the glycoprotein synthesis inhibitor, tunicamycin." Journal of Neurochemistry. 44. 406-410 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kimura M., Mikoshiba K., Tsukada Y., et al.: "Molecular genetic analysis of myelin-deficient mice: Shiverer mutant mice show deletion in gene(s) coding for myelin basic protein." Journal of Neurochemistry. 44. 692-696 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Fujishiro M., Kohsaka S., Tsukada Y., et al.: "Production of monoclonal antibody to 2',3'-cyclic nucleotide 3'-phosphodiesterase from bovine cerebral white matter." Journal of Neurochemistry. 47. 191-195 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Takamiya Y., Kohsaka S., Toya S., et al.: "Possible association of platelet-derive dgrowth factor (PDGF) with the apperrance of reactive astrocytes following bran injury in situ." Brain Research. 383. 305-309 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kohsaka S., Yoshida K., Inoue Y., et al.: "Transplantation of bulk-separated oligodendrocytes into the brains of shiverer mutant mice: Immunohistochemical and electron microscopic studies on the myelination." Brain Research. 372. 137-142 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Fujishiro et.al: J.of Newrocheuistry. 47. 191-195 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Takamiya et.al.: Blain Research. 383. 305-309 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] 高坂,他: 神経研究の進歩. (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] 神経化学. 24. (1985)

    • Related Report
      1985 Annual Research Report
  • [Publications] Journal of Neuro chemistry. (1986)

    • Related Report
      1985 Annual Research Report
  • [Publications] Journal of Neuro chemistry. 44-2. (1985)

    • Related Report
      1985 Annual Research Report

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

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