兔颈上神经节神经元的急性分离及其通道电流记录

Acute isolation of superior cervical ganglion neurons and recording of their ion channel currents in rabbits

目的探讨兔颈上神经节神经元分离及电生理研究的方法。方法为了获得适用于膜片钳实验技术的单个颈上神经节神经细胞，应用酶和机械分离相结合的方法，急性分离20～30d幼兔的单个颈上神经节神经元。通过倒置显微镜直接观察以及全细胞膜片钳技术的电压钳和电流钳分别对分离得钊的颈上神经节神经元的形态学和电生理学特性进行研究。结果急性分离所得活性较好的颈上神经节神经院胞体具有圆形和顶树突特征，立体感强，光晕明显。在全细胞膜片钳记录模式下可记录到全细胞电流、钠电流、钾电流及动作电位。结论该方法可以得到形态和生理特性良好的单个颈上神经节神经元；该方法适用于膜片钳技术的研究，对深入探讨药物、物理因子等对神经节神经元离子通道的影响具有重要的应用价值。

Objective To explore the method of acute isolation of superior cervical ganglion neurons for electrophysiological study. Methods Single superior cervical ganglion neurons suitable for patch-clamp technique were acutely isolated by enzyme digestion and mechanical separation from rabbits of 20-30 days. The morphology and electrophysiologieal characteristics of superior cervical ganglion neurons were investigated through direct observation by inverted microscope and whole-cell patch-clamp technique. Results The superior cervical ganglion neurons obtained by acute isolation were active. Their round cell bodies showed apical dendrite feature. They were three-dimensional and showed obvious halation. Their whole-cell current, sodium current, potassium current and action potential were recorded in whole-cell patch- clamp mode. Conclusions Because single superior cervical ganglion neurons with good morphological and physiological characteristics can be obtained by this method, it is suitable for researches using patch-clamp technique. It is therefore of great value in study of effects of drugs and physical factors on the ion channels of superior cervical ganglion neurons.