Symptomatic dysferlin gene mutation carriers -: Characterization of two cases
Symptomatic dysferlin gene mutation carriers -: Characterization of two cases
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DOI：
10.1212/01.wnl.0000256768.79353.60
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发表时间：
2007-04-17
期刊：
影响因子：
9.9
通讯作者：
Gallardo, E.
中科院分区：
文献类型：
--
作者：
Illa, I.;De Luna, N.;Gallardo, E.
目的：描述两名有症状的 Dysferlin 基因突变携带者。方法：1例患者出现肢带无力。他的兄弟被诊断患有肢带型肌营养不良症 2B，其中 Dysferlin 基因有两个突变（D625Y 和 E1734G）。第二名患者有远端无力。他有两个儿子患有三好肌病，并伴有纯合突变（G519R）。我们使用骨骼肌样本进行了免疫荧光（dysferlin、DAG 蛋白、dysferlin、caveolin-3）、蛋白质印迹（dysferlin、caveolin-3、calpain-3）和实时 PCR（dysferlin）。我们还通过蛋白质印迹研究了外周血单核细胞 (PBM) 中的 Dysferlin。结果：除了肌肉无力外，两名患者还表现出肌酸激酶升高和肌肉 MRI 异常。在筛选整个基因（骨骼肌和单核细胞 mRNA 和基因组 DNA）后，他们仅发现一个等位基因发生突变。肌肉活检标本显示肌膜中度营养不良变化和斑片状dysferlin表达。 PBM 和骨骼肌的蛋白质印迹显示 Dysferlin 显着减少。所有其他蛋白质，包括 Caveolin-3 和 calpain-3 均正常。实时 PCR 显示，与患者受影响的亲属相比，dysferlin mRNA 水平正常。结论：当观察到 Dysferlin 蛋白的病理模式时，应考虑诊断 Dysferlin 突变症状携带者。
Objective: To describe two symptomatic dysferlin gene mutation carriers. Methods: One patient had limb girdle weakness. His brother was diagnosed with limb girdle muscular dystrophy 2B with two mutations in the dysferlin gene (D625Y and E1734G). The second patient had distal weakness. He had two sons with Miyoshi myopathy with a homozygous mutation (G519R). We performed immunofluorescence (dystrophin, DAG proteins, dysferlin, caveolin-3), Western blot (dysferlin, caveolin-3, calpain-3), and real-time PCR (dysferlin) using skeletal muscle samples. We also studied dysferlin in peripheral blood monocytes (PBMs) by Western blot. Results: In addition to the muscle weakness, both patients showed elevated creatine kinase and abnormal muscle MRI. They presented a mutation in only one allele after screening of the whole gene (skeletal muscle and monocyte mRNA and genomic DNA). A muscle biopsy specimen showed moderate dystrophic changes and patchy dysferlin expression in the sarcolemma. Western blot of both PBMs and skeletal muscle demonstrated a significant reduction in dysferlin. All the other proteins including caveolin-3 and calpain-3 were normal. Real-time PCR showed normal levels of dysferlin mRNA vs the patients' affected relatives. Conclusions: The diagnosis of symptomatic carriers of dysferlin mutations should be considered when a pathologic pattern of dysferlin protein is observed.