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NG108-15神经母细胞瘤×胶质瘤细胞内向整流钾电流的特性分析

Characterization of an inward-rectifying potassium current in NG108-15 neuroblastoma x glioma cells.

作者信息

Hu Q, Shi Y L

机构信息

Shanghai Institute of Physiology, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai 200031, Peoples's Republic of China.

出版信息

Pflugers Arch. 1997 Mar;433(5):617-25. doi: 10.1007/s004240050322.

Abstract

By using the whole-cell patch-clamp technique, an inwardly rectifying potassium current, which resembled the "classic" inward-rectifying potassium current (IKIR) of other cells in terms of electrophysiological and pharmacological properties, was identified in db-cAMP-differentiated NG108-15 cells. First, the current was dependent on voltage and time. It could be elicited by applying an initial depolarizing prepulse and a subsequent hyperpolarizing command pulse to the cell. The amplitude of the current depended on both the prepulse and the command pulse and increased with the hyperpolarization of the command pulse as well as the depolarization and the prolongation of the prepulse. The activation and inactivation of the current could be fitted well by single-exponential functions and increased with the hyperpolarization of the membrane. Second, the current was dependent on the extracellular potassium concentration ([K+]o). Elevation of [K+]o resulted in a marked increase in the current amplitude and a positive shift of the peak-current/voltage curve as well as the reversal potential. A tenfold increase of [K+]o introduced an approximately 43-mV shift of the reversal potential, indicating that the current was carried mainly by K+. The conductance (g/gMax) of the current was also dependent on the [K+]o and increased with increases in [K+]o in a manner approximately proportional to the square-root of [K+]o. Finally, the current was sensitive to Cs+ (1 mmol/l), Ba2+ (1 mmol/l) and quinidine (0.2 mmol/l); whereas, two typical potassium channel inhibitors, tetraethylammonium (TEA) and 4-aminopyridine (4-AP), were weak blockers and reduced the current at high concentration (>10 mmol/l). It was also observed that the current was depressed by Cd2+ (1 mmol/l) and Co2+ (1 mmol/l) and increased by perfusing the cell with Ca2+-free solution. Thus, except for the sensitivity to Cd2+, Co2+ and Ca2+, the current displayed most of the hallmarks described for the "classic" IKIR. In conclusion, there appears to be a voltage-dependent IKIR-type inward rectifier in db-cAMP-differentiated NG108-15 cells.

摘要

通过使用全细胞膜片钳技术,在经二丁酰环磷腺苷(db-cAMP)分化的NG108-15细胞中鉴定出一种内向整流钾电流,其在电生理和药理特性方面类似于其他细胞的“经典”内向整流钾电流(IKIR)。首先,该电流依赖于电压和时间。通过向细胞施加初始去极化预脉冲和随后的超极化指令脉冲可引发该电流。电流幅度取决于预脉冲和指令脉冲,并且随着指令脉冲的超极化以及预脉冲的去极化和延长而增加。该电流的激活和失活可用单指数函数很好地拟合,并且随着膜的超极化而增加。其次,该电流依赖于细胞外钾浓度([K+]o)。[K+]o的升高导致电流幅度显著增加,以及峰值电流/电压曲线和反转电位的正向偏移。[K+]o增加十倍会使反转电位产生约43 mV的偏移,表明该电流主要由K+携带。该电流的电导(g/gMax)也依赖于[K+]o,并随着[K+]o的增加而增加,其增加方式与[K+]o的平方根大致成比例。最后,该电流对Cs+(1 mmol/l)、Ba2+(1 mmol/l)和奎尼丁(0.2 mmol/l)敏感;然而,两种典型的钾通道抑制剂,四乙铵(TEA)和4-氨基吡啶(4-AP),是弱阻滞剂,在高浓度(>10 mmol/l)时会降低电流。还观察到该电流受到Cd2+(1 mmol/l)和Co2+(1 mmol/l)的抑制,并且通过用无钙溶液灌注细胞而增加。因此,除了对Cd2+、Co2+和Ca2+的敏感性外,该电流表现出了“经典”IKIR所描述的大多数特征。总之,在经db-cAMP分化的NG108-15细胞中似乎存在一种电压依赖性的IKIR型内向整流器。

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