Goldman L
Department of Physiology, School of Medicine, University of Maryland, Baltimore 21201, USA.
Biophys J. 1995 Dec;69(6):2364-8. doi: 10.1016/S0006-3495(95)80105-0.
Na channel gating parameters in a number of preparations are translated along the voltage axis in excised patches compared to cell attached or whole cell recording. The aim of this study is to determine whether these changes in gating behavior continue over an extended period or, rather, develop rapidly on excision with stationary kinetics thereafter. Average currents were constructed from single-channel records from neuroblastoma N1E 115 at various times after excision, excluding the first 5 min, in eight inside-out excised patches. Single exponentials were fitted to the current decay of the average records, and the mean time constant for each patch was determined. Values were plotted as the percentage difference from these means for each patch against time from excision. Collected results show no obvious trend in values from 5 min to 2 h. Kinetics are stationary, and shifts in Na channel gating parameters along the voltage axis seen in excised as compared to whole cell configuration in neuroblastoma must be complete by the first few minutes after excision. Raising the internal Na concentration reduced the single channel current amplitude, confirming that these are Na channels.
与细胞贴附式或全细胞记录相比,在许多实验制剂中,切除膜片时钠通道门控参数会沿电压轴发生平移。本研究的目的是确定这些门控行为的变化是会在较长时间内持续,还是在切除后迅速发生并随后保持稳定的动力学。在八个内向外膜片切除后的不同时间(不包括最初5分钟),从神经母细胞瘤N1E - 115的单通道记录构建平均电流。将单指数函数拟合到平均记录的电流衰减上,并确定每个膜片的平均时间常数。将每个膜片的值绘制为相对于这些平均值的百分比差异与切除后的时间的关系图。收集的结果显示,从5分钟到2小时的值没有明显趋势。动力学是稳定的,与神经母细胞瘤全细胞配置相比,在切除膜片中观察到的钠通道门控参数沿电压轴的变化在切除后的最初几分钟内必定已经完成。提高内部钠浓度会降低单通道电流幅度,证实这些是钠通道。