Steele J A
Department of Physiology, University of Alberta, Edmonton, Canada.
Pflugers Arch. 1989 Jul;414(3):265-72. doi: 10.1007/BF00584625.
Membrane chloride currents in chick skeletal muscle cells grown in tissue culture were studied by use of the whole cell variation of the patch electrode voltage clamp technique. Small diameter myoballs were obtained by adding colchicine to the growth media. To isolate the currents through the chloride channels, the currents through the sodium, calcium and potassium channels were minimized. With symmetrical chloride concentrations bathing the membrane, inward currents were activated by depolarizations above -45 mV. Above 0 mV, the currents became outward. The reversal potential for the currents shifted with the chloride concentration gradient in a manner consistent with the Nernst relation, indicating that the currents were predominantly carried by chloride ions. The instantaneous current-voltage relation obtained from tail current data was linear. The relationship between conductance and membrane potential was sigmoid. The conductance activated above -45 mV, increased steeply between -45 and -10 mV and saturated above +20 mV. Over the range of potentials where the conductance was just beginning to activate, the conductance increased e-fold for a 7 mV depolarization. The currents activated with an exponential time course and did not decline during step depolarizations. Tail currents declined slowly as the sum of two exponential components. The currents were reversibly suppressed by 100 microM SITS and were irreversibly suppressed by 10 microM DIDS.
运用膜片钳电极电压钳技术的全细胞变体,对组织培养中生长的鸡骨骼肌细胞的膜氯电流进行了研究。通过向生长培养基中添加秋水仙碱获得小直径肌球。为了分离通过氯通道的电流,使通过钠、钙和钾通道的电流最小化。当膜两侧氯浓度对称时,高于 -45 mV 的去极化激活内向电流。高于 0 mV 时,电流变为外向。电流的反转电位随氯浓度梯度变化,其方式与能斯特关系一致,表明电流主要由氯离子携带。从尾电流数据获得的瞬时电流 - 电压关系是线性的。电导与膜电位之间的关系呈 S 形。电导在高于 -45 mV 时激活,在 -45 至 -10 mV 之间急剧增加,在高于 +20 mV 时饱和。在电导刚刚开始激活的电位范围内,电导每去极化 7 mV 增加 e 倍。电流以指数时间进程激活,在阶跃去极化期间不下降。尾电流以两个指数成分之和的形式缓慢下降。电流被 100 microM SITS 可逆抑制,并被 10 microM DIDS 不可逆抑制。