Schofield G G, Ikeda S R
Section of Electrophysiology, National Institute on Alcohol Abuse and Alcoholism, Rockville, MD 20852.
Pflugers Arch. 1988 May;411(5):481-90. doi: 10.1007/BF00582368.
Neurons enzymatically isolated from the adult rat superior cervical ganglion (SCG) were investigated using the whole-cell variant of the patch-clamp technique. Current-clamp studies revealed the following mean passive and active membrane properties: resting membrane potential, -54.9 mV; input resistance, 349 M omega; action potential (AP) threshold, -29.8 mV; AP overshoot, 53.3 mV; AP maximum rate of rise, 166.4 V/s; and AP duration, 3.2 ms. Chemosensitivity to acetylcholine remained intact following enzymatic dispersion. Voltage-clamp studies of a transient tetrodotoxin-sensitive Na+ current revealed activation and inactivation processes which could be fit to modified Boltzmann equations. Na+ current activation parameters for the half activation potential (Vh) and slope factor (K) were -23.3 mV and 5.3 mV, respectively. Inactivation parameters for Vh and K were -59.3 mV and 7.6 mV, respectively. Voltage-clamp studies also revealed a high voltage-activated sustained inward current which was eliminated upon removal of external Ca2+, greatly reduced by 500 microM Cd2+, and supported by Ba2+ or Sr2+. Tail current analysis of this Ca2+ current revealed a sigmoidal activation. A low voltage-activated transient Ca2+ current was not observed. We conclude that isolated SCG neurons retain the properties of neurons in intact ganglia and provide several advantages over conventional preparations for the study of voltage-gated membrane currents.
采用膜片钳技术的全细胞变体,对从成年大鼠颈上神经节(SCG)酶解分离出的神经元进行了研究。电流钳研究揭示了以下平均被动和主动膜特性:静息膜电位为-54.9 mV;输入电阻为349 MΩ;动作电位(AP)阈值为-29.8 mV;AP超射为53.3 mV;AP最大上升速率为166.4 V/s;AP持续时间为3.2 ms。酶解分散后,对乙酰胆碱的化学敏感性保持完整。对瞬时河豚毒素敏感的Na+电流的电压钳研究揭示了可拟合修正玻尔兹曼方程的激活和失活过程。Na+电流激活的半激活电位(Vh)和斜率因子(K)参数分别为-23.3 mV和5.3 mV。Vh和K的失活参数分别为-59.3 mV和7.6 mV。电压钳研究还揭示了一种高电压激活的持续内向电流,去除细胞外Ca2+后该电流消失,500 μM Cd2+可使其大幅降低,而Ba2+或Sr2+可支持该电流。对该Ca2+电流的尾电流分析显示出S形激活。未观察到低电压激活的瞬时Ca2+电流。我们得出结论,分离的SCG神经元保留了完整神经节中神经元的特性,并且在研究电压门控膜电流方面比传统制剂具有几个优势。