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一种改进的适用于膜片钳记录的成年大鼠海马神经元急性分离法

李晓明, 李建国, 杨建明, 胡平, 李晓文, 王颖, 覃鲁宁, 高天明

第一军医大学解剖学教研室.广东,广州 510515；第一军医大学生理学教研室.广东,广州 510515

摘要

该文建立了一种快速、可靠的急性分离成年大鼠海马神经细胞的方法。此法可将实验大鼠的年龄提高到500d以上,体重300g以上;不损伤神经细胞膜的电学特性;形态上有差异的细胞易于分辨。用膜片钳技术的单通道和全细胞模式证实,在该实验条件下,约95%左右的健康细胞均能形成高阻抗封接,并成功地记录了电压依赖性钾、钠、钙通道,外向整流氯通道和大电导的钙激活钾通道电流。

关键词： neuron; patch clamp; hippocampus; rat

An improved method for acute isolation of neurons from the hippocampus of adult rats suitable for patch-clamping study

Li Xiaoming, Li Jianguo, Yang Jianming, Hu Ping, Li Xiaowen, Wang Ying, Qin Luning, Gao Tianming

Department of Anatomy,the First Military Medical University.Guangzhou 510515,Guangdong；China

Abstract

An improved method is described for fast and reliable isolation of neurons from hippocampus of adult rats by a combination of mechanical and enzymatic means. The procedure allows the isolation of neurons from 500--600-d-old rats (over 300 g), preserving the proximal dendritic structure without impairing the electrial characteristics of the cells. Morphologically distinct neurons can be recognized. Using cell-attached, inside-out and whole-cell configurations of patch clamp technique, it was shown that the enzymatically isolated neurons in hippocampus from rats weighing more than 300 g exhibited voltage-gated calcium, sodium and potassium currents, outwardly rectifying chloride channel and large conductance Ca~(2+)-activated potassium channel currents. Approximately, 95% of healthy cells allowed the formatio of giga-ohm seals.

Key words: 神经元;膜片钳;海马;大鼠

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引用本文：

李晓明, 李建国, 杨建明, 胡平, 李晓文, 王颖, 覃鲁宁, 高天明. 一种改进的适用于膜片钳记录的成年大鼠海马神经元急性分离法[J]. 生理学报 2004; 56 (1): .

Li Xiaoming, Li Jianguo, Yang Jianming, Hu Ping, Li Xiaowen, Wang Ying, Qin Luning, Gao Tianming. An improved method for acute isolation of neurons from the hippocampus of adult rats suitable for patch-clamping study. Acta Physiol Sin 2004; 56 (1): (in Chinese with English abstract).