Shuyun Huang , Qing Cai , Weitian Liu , Xiaoling Wang , Tao Wang
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引用次数: 2
摘要
目的记录急性离体海马锥体神经元钙、钾、钠电流。方法海马CA3神经元用1 mg蛋白酶/3 ml SES分离，并用尖端直径逐渐变小的抛光移液管进行机械培养。采用全细胞模式膜片钳技术记录电压门控通道电流。结果该方法分离了海马神经元，保留了顶端树突和几个基底树突，但未损害神经元的电特性。全细胞膜片钳结构被成功地用于记录电压门控Ca2+电流、延迟整流K+电流和电压门控Na+电流。结论蛋白酶联合机械磨蚀可用于大鼠海马神经元的解离。电压门控通道电流可以使用膜片钳技术进行记录。
Whole-cell recordings of voltage-gated Calcium, Potassium and Sodium currents in acutely isolated hippocampal pyramidal neurons
Objective
To record Calcium, Potassium and Sodium currents in acutely isolated hippocampal pyramidal neurons.
Methods
Hippocampal CA3 neurons were freshly isolated by 1 mg protease/3 ml SES and mechanical trituration with polished pipettes of progressively smaller tip diameters. Patch clamp technique in whole-cell mode was employed to record voltage-gated channel currents.
Results
The procedure dissociated hippocampal neurons, preserving apical dendrites and several basal dendrites, without impairing the electrical characteristics of the neurons. Whole-cell patch clamp configuration was successfully used to record voltage-gated Ca2+ currents, delayed rectifier K+ current and voltage-gated Na+ currents.
Conclusion
Protease combined with mechanical trituration may be used for the dissociation of neurons from rat hippocampus. Voltage-gated channels currents could be recorded using a patch clamp technique.
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