小鼠海马神经元急性分离和膜片钳技术的应用

为了获得适用于膜片钳实验技术的单个海马神经元，应用酶消化及机械分离法，急性分离出生10～13d的昆明小鼠得到单个海马神经元通过倒置显微镜直接观察和全细胞膜片钳技术分别对分离得到的海马细胞的形态学和电生理学特征进行研究．通过实验可得，急性分离所得活性较好的海马神经元胞体具有锥形胞体和顸树突特征，立体感强．在全细胞膜片钳记录模式下可记录到全细胞电流及钠、钾通道电流．由实验结果可知，该方法可以得到形态和生理特征良好的单个海马神经元，证实该方法适用于膜片钳技术的研究，对深入探讨药物、物理因子等对海马离子通道及信号转导机制的作用有重要的应用价值．

Acute Isolation of Hippocampal Neurons of Neonate Mice and Application of Patch-Clamp Technique

To gain a single hippocampal neuron suitable for patch-clamp technique, acute isolation of hippocampal neurons of 10-13-d-old Kunming mouse was obtained by enzyme digestion and mechanical separation tech＇hique. With the direct observation by inverted microscope and whole-cell patch-clamp technique, the morphology and electrophysiology characteristics of hippocampal cells were investigated respectively. The experimental results show that hippocampal neurons with better activity obtained by acute isolation are of cone-cell body and apical dendrite, three-dimensional sense. The whole-cell current and sodium current as well as potassium current can be recorded under whole-cell patch-clamp mode. The single hippocampal neuron with good characteristics in both morphology and physiology can be obtained with this method, which is suitable for the research of patch-clamp technique and provides an important way to further study the effects of drugs and physical factors on the ion channels of hippocampal neurons and the mechanism of signal transduction.