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小鼠巨噬细胞系J774.1中的内向整流全细胞和单通道钾电流

Inwardly rectifying whole-cell and single-channel K currents in the murine macrophage cell line J774.1.

作者信息

McKinney L C, Gallin E K

机构信息

Department of Physiology, Armed Forces Radiobiology Research Institute, Bethesda, Maryland 20814-5145.

出版信息

J Membr Biol. 1988 Jul;103(1):41-53. doi: 10.1007/BF01871931.

Abstract

Inward currents in the murine macrophage-like cell line J774.1 were studied using the whole-cell and cell-attached variations of the patch-clamp technique. When cells were bathed in Na Hanks' (KCl = 4.5 mM, NaCl = 145 mM), and the electrode contained Na-free K Hanks' (KCl = 145 mM) single-channel currents were observed at potentials below -40 mV which showed inward rectification, were K-selective, and were blocked by 2.5 mM Ba in the pipette. Single-channel conductance was 29 pS, and was proportional to the square root of [K]o. Channels manifested complex kinetics, with multiple open and closed states. The steady-state open probability of the channel was voltage dependent, and declined from 0.9 to 0.45 between -40 and -140 mV. When hyperpolarizing voltage pulses were repetitively applied in the cell-attached patch mode, averaged single-channel currents showed inactivation. Inactivation of inwardly rectifying whole-cell current was measured in Na Hanks' and in two types of Na-free Hanks': one with a normal K concentration (4.5 mM) and the other containing 145 mM K. Inactivation was shown to have Na-dependent and Na-independent components. Properties of single-channel current were found to be sufficient to account for the behavior of the macroscopic current, except that single-channel current showed a greater degree of Na-independent inactivation than whole-cell current.

摘要

利用膜片钳技术的全细胞和细胞贴附模式,研究了小鼠巨噬细胞样细胞系J774.1中的内向电流。当细胞浸浴在钠汉克斯液(KCl = 4.5 mM,NaCl = 145 mM)中,且电极内含有无钠钾汉克斯液(KCl = 145 mM)时,在低于-40 mV的电位下观察到单通道电流,该电流表现出内向整流、对钾离子有选择性,并且被移液管中的2.5 mM Ba阻断。单通道电导为29 pS,并且与[K]o的平方根成正比。通道表现出复杂的动力学,具有多个开放和关闭状态。通道的稳态开放概率依赖于电压,在-40至-140 mV之间从0.9降至0.45。当在细胞贴附膜片模式下重复施加超极化电压脉冲时,平均单通道电流显示出失活。在钠汉克斯液以及两种无钠汉克斯液中测量了内向整流全细胞电流的失活:一种具有正常钾浓度(4.5 mM),另一种含有145 mM K。结果表明失活具有钠依赖性和非钠依赖性成分。发现单通道电流的特性足以解释宏观电流的行为,只是单通道电流显示出比全细胞电流更大程度的非钠依赖性失活。

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