| Project/Area Number |
06670671
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurology
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| Research Institution | Tokai University |
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Principal Investigator |
YOSHII Fumihito Tokai University, Neurology, Assistant Prof., 医学部, 講師 (90130103)
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| Co-Investigator(Kenkyū-buntansha) |
ONOE Kyuichiro Tokai University, Neurology, Assistant, 医学部, 助手 (50276812)
YOSHITOSHI Mieko Tokai University, Neurology, Assistant, 医学部, 助手 (50183702)
OHASUGA Hitoshi Tokai University, Neurology, Assistant, 医学部, 助手 (60203775)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1994: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | myotonic dystrophy / trinucleotide repeat / PCR method / myotonin protein kinase / immunohistochemical staining / ion-channel / patch clamp technique / 3.塩基反復配列 / 筋緊張性ジストロフィー / CTGリピート / 細胞膜 / DMキナーゼ / 免疫組織化学 |
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| Research Abstract |
The purposes of the study are to discover the relationship between the variety of clinical features in myotonic dystrophy (DM) and DM gene abnormality, and between DM gene abnormality and the changes in ion-channel currents of muscular cell membrane in DM patients.
1. DNA analysis in myotonic dystrophy
Using the PCR method, we studied an expanded trinucleotide (CTG) repeat in a gene on 19 chromosome in patients with DM and healthy subjects. We found two bands on agarose gel in healthy subjects, but only one band appeared in patients with DM, probably due to enormous expansion of CTG repeat in the abnormal allele. For further evaluation we intend to use a specific probe for Southern blotting to study the exact repeat number of CTG in DM patients.
2. Localization of myotonin protein kinase (DM-PK) in muscular cells
Using rabbit anti-DM-PK polyclonal antibodies and indirect immunohistochemical staining method, we studied the localization of DM-PK in the muscular cells. Brownish reactive substances were observed along the inner side of the cell membrane in normal muscular cells, but no substances were observed in the muscular cells of the patients with DM.In the muscular cells of myositis, there were many reactive substances, especially in the cytoplasm of the regenerating muscular cells.
3. Ion-channel currents of the muscular cell membrane
Ion-channel currents of the muscular cell membrane in patients with DM were measured by patch-clamp method. However, it failed to measure exactly the currents in mononuclear myoblasts or polynuclear muscular cells. We are now doing basic studies using muscular specimens of the animals.
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