DeCoursey T E
Department of Physiology, Rush Presbyterian St. Luke's Medical Center, Chicago, Illinois 60612.
Biophys J. 1991 Nov;60(5):1243-53. doi: 10.1016/S0006-3495(91)82158-0.
Alveolar epithelial cells isolated from rats and maintained in primary culture were studied using the whole-cell configuration of the "patch-clamp" technique. After other ionic conductances were eliminated by replacing permeant ions with N-methyl-D-glucamine methanesulfonate, large voltage-activated hydrogen-selective currents were observed. Like H+ currents in snail neurons and axolotl oocytes, those in alveolar epithelium are activated by depolarization, deactivate upon repolarization, and are inhibited by Cd2+ and Zn2+. Activation of H+ currents is slower in alveolar epithelium than in other tissues, and often has a sigmoid time course. Activation occurs at more positive potentials when external pH is decreased. Saturation of the currents suggests that diffusion limitation may occur; increasing the pipette buffer concentration from 5 to 120 mM at a constant pH of 5.5 increased the maximum current density from 8.7 to 27.3 pA/pF, indicating that the current amplitude can be limited in 5 mM buffer solutions by the rate at which buffer molecules can supply H+ to the membrane. These data indicate that voltage-dependent H+ currents exist in mammalian cells.
使用“膜片钳”技术的全细胞模式,对从大鼠中分离并维持在原代培养中的肺泡上皮细胞进行了研究。在用甲磺酸N-甲基-D-葡糖胺替代通透离子消除其他离子电导后,观察到了大的电压激活的氢选择性电流。与蜗牛神经元和蝾螈卵母细胞中的H⁺电流一样,肺泡上皮中的H⁺电流由去极化激活,复极化时失活,并受Cd²⁺和Zn²⁺抑制。肺泡上皮中H⁺电流的激活比其他组织慢,且通常具有S形的时间进程。当外部pH降低时,在更正的电位下发生激活。电流的饱和表明可能发生扩散限制;在pH 5.5恒定的情况下,将移液管缓冲液浓度从5 mM增加到120 mM,最大电流密度从8.7 pA/pF增加到27.3 pA/pF,这表明在5 mM缓冲液中,电流幅度可能受缓冲分子向膜供应H⁺的速率限制。这些数据表明哺乳动物细胞中存在电压依赖性H⁺电流。