Department of Neurology and Center for Neuroscience, University of California, Davis, CA 95616, USA.
Neuromolecular Med. 2012 Dec;14(4):328-37. doi: 10.1007/s12017-012-8190-1. Epub 2012 Jul 12.
Myotonia congenita-inducing mutations in the muscle chloride channel CLC-1 normally result in reduced open probability (P (o)) of this channel. One well-accepted mechanism of the dominant inheritance of this disease involves a dominant-negative effect of the mutation on the function of the common-gate of this homodimeric, double-barreled molecule. We report here a family with myotonia congenita characterized by muscle stiffness and clinical and electrophysiologic myotonic phenomena transmitted in an autosomal dominant pattern. DNA sequencing of DMPK and ZNF9 genes for myotonic muscular dystrophy types I and II was normal, whereas sequencing of CLC-1 encoding gene, CLCN1, identified a single heterozygous missense mutation, G233S. Patch-clamp analyses of this mutant CLC-1 channel in Xenopus oocytes revealed an increased P (o) of the channel's fast-gate, from ~0.4 in the wild type to >0.9 in the mutant at -90 mV. In contrast, the mutant exhibits a minimal effect on the P (o) of the common-gate. These results are consistent with the structural prediction that the mutation site is adjacent to the fast-gate of the channel. Overall, the mutant could lead to a significantly reduced dynamic response of CLC-1 to membrane depolarization, from a fivefold increase in chloride conductance in the wild type to a twofold increase in the mutant-this might result in slower membrane repolarization during an action potential. Since expression levels of the mutant and wild-type subunits in artificial model cell systems were unable to explain the disease symptoms, the mechanism leading to dominant inheritance in this family remains to be determined.
先天性肌强直诱导突变通常会导致肌肉氯离子通道 CLC-1 的开放概率(P(o))降低。这种疾病的显性遗传的一种公认机制涉及突变对这种同二聚体、双桶分子的公共门控的功能的显性负效应。我们在这里报告一个先天性肌强直家族,其特征为肌肉僵硬和临床及电生理肌强直性现象呈常染色体显性遗传模式。肌强直性肌营养不良 I 型和 II 型的 DMPK 和 ZNF9 基因的 DNA 测序正常,而 CLC-1 编码基因 CLCN1 的测序则鉴定出单个杂合错义突变 G233S。在爪蟾卵母细胞中对这种突变 CLC-1 通道进行的膜片钳分析显示,通道的快速门控的 P(o)增加,从野生型的约 0.4 增加到突变型的 -90 mV 时的>0.9。相比之下,突变对公共门控的 P(o)几乎没有影响。这些结果与结构预测一致,即突变位点紧邻通道的快速门控。总的来说,突变可能导致 CLC-1 对膜去极化的动态反应显著降低,从野生型中氯离子电导的五倍增加到突变型中的两倍增加 - 这可能导致动作电位期间膜复极化变慢。由于人工模型细胞系统中突变型和野生型亚基的表达水平无法解释疾病症状,因此该家族中导致显性遗传的机制仍有待确定。