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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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Citing This Article：

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).