| 刘 超,肖 乐,段现来,王爱民,曾乐平.全面性癫痫伴热性惊厥附加症SCN1A基因G302D突变的膜片钳电生理研究[J].现代生物医学进展英文版,2019,19(18):3461-3464. |
| 全面性癫痫伴热性惊厥附加症SCN1A基因G302D突变的膜片钳电生理研究 |
| Functional Analysis of a Novel SCN1A Mutation G302D Identified in GEFS+ |
| Received:June 23, 2019 Revised:July 18, 2019 |
| DOI：10.13241/j.cnki.pmb.2019.18.012 |
| 中文关键词: 全面性癫痫伴热性惊厥附加症 SCN1A基因 突变 钠离子通道 |
| 英文关键词: Generalized epilepsy with febrile seizures plus SCN1A gene Mutations Sodium ion channel |
| 基金项目:湖南省科技计划项目（2016JC2054）；长沙市科技计划项目（kq1706001） |
| Author Name | Affiliation | | LIU Chao | The first hospital of changsha, Changsha, Hunan, 410005, China | | XIAO Le | The first hospital of changsha, Changsha, Hunan, 410005, China | | DUAN Xian-lai | The third hospital of changsha, Changsha, Hunan, 410005, China | | WANG Ai-min | The first hospital of changsha, Changsha, Hunan, 410005, China | | ENG Le-ping | Xiangya School of Medicine, Central South University, Changsha, Hunan, 410013, China |
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| 中文摘要: |
| 摘要 目的：探讨全面性癫痫伴热性惊厥附加症（GEFS+）SCN1A基因G302D突变的电生理机制。方法：采用体外定点诱变法构建携带有基因突变G302D的pCMV-SCN1A的表达载体，lipo2000脂质体转染法共转染pCMV-SCN1A-G302D质粒和pCD8-IRES-SCN1B质粒到HEK-293细胞系，进行全细胞膜片钳电生理实验记录Navl.1通道电流及动力学参数，由pClamp10.0以及OriginPro8.0软件分析。结果：SCN1A-G302D 突变体与野生型相比，电流密度降低，激活速度减慢，失活后恢复时间延长。失活参数两者相比没有显著性统计学差异。结论：SCN1A基因G302D突变导致Navl.1通道功能部分丧失，可能是导致GEFS+的病因。 |
| 英文摘要: |
| ABSTRACT Objective: To elucidate the molecular and electrophysiological mechanisms of generalized epilepsy with febrile seizures plus through functional analysis of a novel SCN1A gene mutation G302D. Methods: A recombinant plasmid pCMV-SCN1A containing the mutant G302D was constructed by the PCR-based site-directed mutagenesis technique. Lipofectamine 2000 was used to co-transfect the plasmid pCMV-SCN1A-G302D and pCD8-IRES-SCN1B into HEK-293 cells line to induce stable expression of α1 and β1 subunit of Na+ channel. During patch clamp detection, the standard stimulus was given in the whole-cell voltage clamp mode. Data acquisition and analysis were accomplished with PatchMaster10.0 and OriginPro8.0. Results: SCN1A-G302D mutants displayed a decreased current density, a slower activation and a significantly delayed recovery from inactivation. There were no significantly differences in the steady-state inactivation between two groups. Conclusion: These data indicates that SCN1A-G302D mutants result in partial loss of function, which may be the cause of GEFS +. |
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