Ypey D L, Clapham D E
Proc Natl Acad Sci U S A. 1984 May;81(10):3083-7. doi: 10.1073/pnas.81.10.3083.
Patch clamp techniques were used to study ionic currents in cultured mouse peritoneal macrophages. Whole-cell voltage clamp studies of cells 1-5 hr after isolation showed only a high-resistance linear membrane. After 1 day in culture, 82 of 85 cells studied had developed a voltage- and time-dependent potassium (K+) conductance similar to the delayed outward rectifier in nerve and muscle cells. The current activated when the membrane was depolarized above -50 mV. The sigmoidally rising current rose to a peak at a rate that increased with depolarization. Inactivation proceeded exponentially with a time constant of approximately equal to 450 ms. Recovery from inactivation was slow (tau = 12 s). The reversal potentials for varying extracellular K+ concentrations followed the Nernst predictions for a K+ -specific channel. The conductance was blocked by extracellular 4-aminopyridine and by intracellular tetraethylammonium chloride, barium, and cesium. Single-channel K+ currents comprising this net current had a conductance of 16 pS, exhibited bursting behavior, and inactivated with time. No inward currents were ever detected in macrophages cultivated for up to 4 days. Short-term exposure to chemoattractant and transmitter agents failed to activate an inward current. Macrophages may change their membrane electrophysiological properties depending on their state of functional activation. We postulate that the K+ conductance develops prior to depolarizing conductances involved in the macrophage's immunological functions.
采用膜片钳技术研究培养的小鼠腹腔巨噬细胞中的离子电流。对分离后1 - 5小时的细胞进行全细胞膜片钳研究,结果显示仅存在高电阻线性膜。培养1天后,所研究的85个细胞中有82个细胞出现了电压和时间依赖性钾(K +)电导,类似于神经和肌肉细胞中的延迟外向整流器。当膜电位去极化至 - 50 mV以上时,电流被激活。呈S形上升的电流以随去极化增加的速率升至峰值。失活呈指数形式进行,时间常数约为450毫秒。失活后的恢复缓慢(时间常数τ = 12秒)。不同细胞外K +浓度下的反转电位符合针对K +特异性通道的能斯特预测。该电导被细胞外4 -氨基吡啶以及细胞内氯化四乙铵、钡和铯阻断。构成该净电流的单通道K +电流电导为16 pS,表现出爆发行为,并随时间失活。在培养长达4天的巨噬细胞中从未检测到内向电流。短期暴露于趋化因子和递质未能激活内向电流。巨噬细胞可能会根据其功能激活状态改变其膜电生理特性。我们推测K +电导在参与巨噬细胞免疫功能的去极化电导之前就已形成。