| Project/Area Number |
60870032
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurology
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| Research Institution | KEIO UNIVERSITY SCHOOL OF MEDICINE |
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Principal Investigator |
TSUKADA Yasuzo KEIO UNIV. SCHOOL OF MED., PROFESSOR, 医学部, 教授 (00050956)
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| 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
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| 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)
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| Keywords | Monoclonal 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 |
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| 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.
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