Lee S C, Levy D I, Deutsch C
Department of Physiology, University of Pennsylvania, Philadelphia 19104-6085.
J Gen Physiol. 1992 May;99(5):771-93. doi: 10.1085/jgp.99.5.771.
We used patch clamp techniques to identify and characterize a variety of K+ channels in primary human peripheral T lymphocytes. The most common channel observed in cell-attached configuration was voltage gated and inactivating. In ensemble averages, the kinetics of its activation and inactivation were similar to those of the whole-cell, voltage-gated K+ current described previously (Cahalan, M. D., K. G. Chandy, T. E. DeCoursey, and S. Gupta. 1985. J. Physiol. [Lond.]. 358:197-237; Deutsch, C., D. Krause, and S. C. Lee. 1986. J. Physiol. [Lond.]. 372:405-423), suggesting that this channel underlies the major portion of the outward current in lymphocytes. A small fraction of the time, this or another very similar channel was observed to inactivate significantly more slowly. Another channel type observed in cell-attached recording was seen less frequently and was transient in its appearance. This channel has a unitary conductance of approximately 10 pS, similar to the voltage-gated channel, but its voltage-independent gating, lack of inactivation, and different kinetic parameters showed it to be distinct. In whole-cell recording there is often a significant plateau current during sustained depolarization. Experiments using whole-cell and excised outside-out configurations indicate that at least part of this residual current is carried by K+ and, as opposed to the predominant voltage-gated current, is charybdotoxin insensitive. These findings are consistent with evidence that implicates charybdotoxin-sensitive and -insensitive components in T lymphocyte proliferation and volume regulation.
我们运用膜片钳技术来识别和表征原代人外周血T淋巴细胞中的多种钾离子通道。在细胞贴附模式下观察到的最常见通道是电压门控且可失活的。在整体平均值中,其激活和失活的动力学与先前描述的全细胞膜片钳电压门控钾离子电流相似(Cahalan, M. D., K. G. Chandy, T. E. DeCoursey, and S. Gupta. 1985. J. Physiol. [Lond.]. 358:197 - 237; Deutsch, C., D. Krause, and S. C. Lee. 1986. J. Physiol. [Lond.]. 372:405 - 423),这表明该通道是淋巴细胞外向电流的主要组成部分。在一小部分时间里,观察到这个或另一个非常相似的通道失活明显更慢。在细胞贴附记录中观察到的另一种通道类型出现频率较低且呈瞬时性。该通道的单位电导约为10 pS,与电压门控通道相似,但它的电压非依赖性门控、无失活以及不同的动力学参数表明它是不同的。在全细胞记录中,持续去极化期间通常会有显著的平台电流。使用全细胞和外翻式膜片钳配置的实验表明,至少部分这种残余电流是由钾离子携带的,并且与主要的电压门控电流不同,它对蝎毒素不敏感。这些发现与涉及蝎毒素敏感和不敏感成分参与T淋巴细胞增殖和体积调节的证据一致。