| Project/Area Number |
07670891
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Jichi Medical School |
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Principal Investigator |
KOBAYASHI Yoko Jichi Medical School, Department of Pediatrics Assistant, 医学部, 助手 (20245046)
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| Co-Investigator(Kenkyū-buntansha) |
SAITO Shigeko Jichi Medical School, Department of Pediatrics Assistant, 医学部, 助手 (00260836)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1995: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | LHON / MELAS / DNA mitochondirial / Mitochondrial genetics / Polymerase chain reaction / mitochondrial DNA / mitochondrial myopathy / heteroplasmy / PCR / DNA, -Mitochondrial-genetics / Point-Mutation / Mitochondrial-Myopathies-genetic / MELAS-Syndrome-genetics |
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| Research Abstract |
We have examined the distribution of mutant mtDNA molecules in single cells from a patient with Leber hereditary optic neuropathy (LHON) and MELAS (mitochondrial encephalopathy, myopathy, lactic acidosis and stroke-like episodes). The patient was a 46-year-old unaffected male in a large pedigree with LHON.Previous analysis of DNA from the total blood buffy coat had shown that -80 % of the mtDNA molecules carried the 11778 LHON mutation and that 20 % had the normal sequence at this point. Single lymphocytes were isolated using a micromanipulation method. Each isolated cell was used for two step PCR amplification and restriction digestion. Twenty-five samples each containing a single cell were analyzed. Sixteen cells had completely mutant mtDNA,and five cells had completely normal mtDNA (intracellular homoplasmy). Four cells contained a mixture of both. Twenty one of the 25 cells had segregated to either homoplasmic mutant or homoplasmic normal mtDNA.mtDNA heteroplasmy in peripheral lymphocytes in this patient therefore was intercellular to a large extent. However, almost all cells which showed 95 % normal mitochondrial genes in MELAS rapidly changed to the cells involved mutant genes in the homoplasmic fashion after cell culture.
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