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 Mg2+-free external
沈丘月 1刘娜娜 2刘黎黎 1姜玉武 2侯新琳1
作者信息
- 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(北京 100034);儿科遗传性疾病分子诊断与研究北京市重点实验室
- 折叠
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.
Abstract
Objective To investigate the electrophysiological characteristics of hamster primary cortical neurons by using the whole-cell patch-clamp recordings in a Mg2+-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(Mg2+group)or Mg2+-free external(Mg2+-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 Mg2+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 Mg2+-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 Mg2+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Key words
Epilepsy/Syrian hamsters/Cerebral cortex/Cortical excitability/Neuron/Patch-clamp techniques/Ex-citatory postsynaptic current/Excitatory postsynaptic potential引用本文复制引用
基金项目
国家自然科学基金(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(2021YFC2700700)
出版年
2024
NETL
NSTL
万方数据