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目的研究儿童失神癫痫(childhoodabsenceepilepsy,CAE)患儿CACNA1H基因G773D突变对钙通道功能的影响。方法用定点突变重叠延伸聚合酶链反应(polymerasechainreaction,PCR)方法构建G773D突变体,脂质体法将突变体和野生型人Cav3.2acDNA分别转染HEK-293细胞,获得稳定表达细胞株,全细胞膜片钳法研究其电生理变化。结果突变体和野生型细胞钙通道激活和失活动力学差异无统计学意义,但突变体G773D钙电流密度明显高于野生型。结论CACNA1H基因G773D突变可使其编码通道电流增加,并可能引起神经元兴奋性增加。
Objective To investigate the effect of G773D mutation in CACNA1H gene on calcium channel function in children with childhood absent epilepsy (CAE). Methods The G773D mutant was constructed by polymerase chain reaction (PCR) with site-directed mutagenesis. The mutant and wild-type human Cav3.2 cDNA were transfected into HEK-293 cells by lipofectamine. The stable cell lines were obtained. Cell patch clamp study of its electrophysiological changes. Results The activation and inactivation kinetics of calcium channels in mutant and wild-type cells were not statistically different. However, the G773D calcium current density was significantly higher than that in wild type. Conclusion G773D mutation of CACNA1H gene can increase the current of its encoding pathway and may result in the increase of excitability of neurons.