| Project/Area Number |
16591052
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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 | Saitama Medical University |
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Principal Investigator |
OHTAKE Akira Saitama Medical University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (00203810)
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| Co-Investigator(Kenkyū-buntansha) |
SASAKI Nozomu Saitama Medical University, Faculty of Medicine, Professor, 医学部, 教授 (00009672)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Congenital lactic acidosis / Blue native polyacrylamide gel electrophoresis / Skin fibroblast / Mitochondrial respiratory chain / Immunoblotting / In gel enzyme staining / Blood lactate / pyruvate ratio / ミトコンドリア呼吸鎖異常症 / 血中Lactate / Pyruvate比 / Blue Native電気泳動(BN PAGE) |
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| Research Abstract |
Mitochondrial respiratory chain disorders are not only the most common but the most clinically diverse group of inborn errors of metabolism. It is often difficult to establish an accurate diagnosis and then to determine the molecular basis of respiratory chain disorders. Blue native polyacrylamide gel electrophoresis (BN-PAGE) is a form of native electrophoresis for resolving membrane protein complexes. We used BN-PAGE combined either with in gel enzyme staining or with immunoblotting to analyze 25 patients with possible, probable or definite mitochondrial respiratory chain disorders as defined by the criteria of Bernier et al. A potential complex I abnormality was detected in 13 out of 25 patients, but no abnormalities were found either in the amounts or activities of other complexes. Complex I abnormalities were categorized into three groups based on amount and size of assembled complex I. Combined with conventional enzyme assay, 4 out of 13 patients having complex I abnormality in BN-PAGE analyses were diagnosed as definite complex I deficiency. We conclude that BN-PAGE is a powerful tool both in the correct diagnosis and as a useful guide to future molecular analysis for respiratory chain disorders.
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