大鼠海马CAⅠ区锥体细胞的急性分离及全细胞记录方法

Methods of acute isolation of CAⅠ pyramidal neurons in hippocampus of neonate rats and whole-cell recording

目的寻求一种适用于膜片钳技术(PCT)的大鼠海马CAⅠ区锥体细胞的急性分离及全细胞记录方法。方法采用酶加机械分离的方法制备10～16d鼠龄的海马CAⅠ区锥体神经元。其电生理学特性测定用全细胞PCT。结果所分离的85%神经元不仅形态学特征保存完好,而且保存了主要的离子通道活性。结论成功建立了一种简单、快速、可靠的适用于PCT的大鼠皮层神经元急性分离方法。

Objective To establish a method of acute isolation of neonate rat hippocampal CA Ⅰ pyramidal neurons suitable for patch clamp technique （PCT）.Methods Neonate rat hippocampal CA Ⅰ pyramidal neurons were obtained by a combination of mechanical and enzymatic means.The electrophysiologic properties were detected by the whole-cell patch clamp technique.Results 85% of isolated neurons exhibited the well-kept proximal dendritic structures with clean surface, and preserved activities of typical ion channels. Conclusions A simple, fast and credible method of acute isolation of neonate rat hippocampal CA Ⅰ pyramidal neurons suitable for whole-cell recording is successfully founded.