培养的大脑皮层、海马及交感神经细胞钠、钾和钙离子通道的全细胞记录技术  

WHOLE-CELL RECORDING OF POTASSIUM AND SODIUM CURRENTS IN CORTICAL,HIPPOCAMPAl,AND SYMPATHETIC GANGLIA NEURONS

作　　者：崔文玉[1] 张颖丽[1] 殷晓峰[1] 汪海[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《中国应用生理学杂志》2005年第1期105-110,共6页Chinese Journal of Applied Physiology

基　　金：军队医药卫生"十五"规划重点课题 (0 2Z0 2 5 )

摘　　要：目的 :建立新生大鼠大脑皮层、海马细胞及交感神经元细胞的培养方法及其钠、钾和钙通道的膜片钳全细胞记录技术。方法 :取出生 1～ 3d的大鼠大脑皮层、海马及交感神经节 ,用胰蛋白酶 (0 .12 5 % )消化组织并分离出神经细胞 ,种植在涂有多聚赖氨酸的 35mm培养皿中 ,用高糖的DMEM培养液培养 ,一周后 ,镜下可见神经细胞壁光滑、完整 ,周围有明亮的光润 ,在高倍倒置显微镜下可见到完整的细胞核及均匀的胞浆 ,细胞间形成良好的突触连接 ,可用于细胞膜片钳记录。结果 :用胰蛋白酶消化分离培养的神经细胞 ,功能状态良好 ,在膜片钳全细胞记录中 ,易形成细胞与记录电极间的高阻抗封接 ,可分别记录到INa、IA、IK 和ICa。结论 :在神经系统电生理学研究中 ,此方法可应用于中枢神经系统不同脑区培养以及钠、钾和钙通道电流的记录。Aim: To establish the culturing of cortical, hippocampal and sympathetic ganglia neurons derived from neonatal rats and the whole-cell recording techniques of potassium and sodium currents in cultured neurons. Methods: Cortex, hippocampus and sympathetic ganglion were isolated from neonatal rats(1～3 days) and digested by trypsin(0.125%). The neurons were cultured in polysine covered petri dish(35 mm) with DEME medium(high glucose) for seven days for electrophysiological experiments. Results: The neurons cultured by this method were in good condiditons and could be sealed easily in whole-cell recording. I Na , I A, and I K could be recorded respectively in cultured neurons by different recording paradigms in Ca 2+ -free extracellular solution. Conclusion: It is a practicable and economical method for electrophysiological study of nervous system.

关 键 词：膜片钳 皮层 海马 交感神经元 钾电流 钠电流 钙电流 

分 类 号：Q421[生物学—神经生物学]