无镁诱导仓鼠原代皮质神经元电生理学特性  

The electrophysiological characteristics of primary hamster cortical neurons with Mg^(2+)-free external

作　　者：沈丘月 刘娜娜 刘黎黎[1] 姜玉武[1,2] 侯新琳[1] SHEN Qiuyue;LIU Nana;LIU Lili;JIANG Yuwu;HOU Xinlin(Department of Pediatrics,Peking University First Hospital,Beijing 100034,China)

机构地区：[1]北京大学第一医院儿科,北京100034 [2]北京大学第一医院儿科遗传性疾病分子诊断与研究北京市重点实验室,北京100034

出　　处：《中国神经精神疾病杂志》2024年第3期140-145,共6页Chinese Journal of Nervous and Mental Diseases

基　　金：国家自然科学基金项目(编号:82101806);国家重点研发计划生育健康及妇女儿童健康保障重点专项(编号:2021YFC2700700)。

摘　　要：目的无镁细胞外液建立癫痫放电模型,利用全细胞膜片钳技术,检测仓鼠原代皮质神经元的电生理学特性。方法采用生后1~2 d新生叙利亚仓鼠,分离大脑皮质培养原代神经元培养至第12天,分别予有镁细胞外液(有镁组)和无镁细胞外液(无镁组)孵育3 h,3 h后均更换为正常孵育液继续培养24 h。利用全细胞膜片钳技术,电压钳模式下记录神经元兴奋性突触后电流(excitatory postsynaptic currents,EPSC),电流钳模式下记录神经元兴奋性突触后电位(excitatory postsynaptic potentials,EPSP)。结果与有镁组相比,无镁组仓鼠原代皮质神经元EPSC[(124.38±75.15)Hz vs.(33.93±22.32)Hz,P<0.001]及EPSP[(37.05±38.37)Hz vs.(5.63±9.52)Hz,P<0.01]的频率升高,且差异有统计学意义;而两组之间EPSC及EPSP的振幅、曲线下面积和半宽度的差异无统计学意义(P>0.05)。结论无镁处理后仓鼠原代皮质神经元兴奋性升高,仓鼠原代皮质神经元可用于构建癫痫细胞模型。Objective To investigate the electrophysiological characteristics of hamster primary cortical neurons by using the whole-cell patch-clamp recordings in a Mg^(2+)-free cell model of epilepsy.Methods The cerebral cortex were isolated for primary neuron culture from neonatal Syrian hamsters 1-2 days after birth.After 12 days,primary neurons were exposed to normal external(Mg^(2+)group)or Mg^(2+)-free external(Mg^(2+)-free group)respectively for 3 hours,followed by a 24-hour incubation in normal culture medium.Excitatory postsynaptic currents(EPSC)and excitatory postsynaptic potentials(EPSP)were recorded in voltage clamp mode and current clamp mode using whole-cell patch-clamp recordings,respectively.Results Compared to Mg^(2+)group,the frequency of EPSC[(124.38±75.15)Hz vs.(33.93±22.32)Hz,P<0.001](P<0.001)and EPSP[(37.05±38.37)Hz vs.(5.63±9.52)Hz,P<0.01]in primary cortical neurons from the Mg^(2+)-free group increased significantly.Meanwhile,there were no statistically significant differences in the amplitude,area under the curve and half-width of EPSC and EPSP between the two groups(P>0.05).Conclusion The present study demonstrates an increase in excitability of primary hamsters cortical neurons after Mg^(2+)depletion,suggesting that these neurons can be utilized to use to create cell models of epilepsy.

关 键 词：癫痫 叙利亚仓鼠 大脑皮质 皮质兴奋性 神经元 膜片钳技术 兴奋性突触后电流 兴奋性突触后电位 

分 类 号：R742.1[医药卫生—神经病学与精神病学] R338[医药卫生—临床医学]