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离体美西螈肠上皮细胞的钾电流:一项全细胞膜片钳研究。

Potassium currents of isolated Necturus enterocytes: a whole-cell patch-clamp study.

作者信息

Sheppard D N, Valverde M A, Giraldez F, Sepúlveda F V

机构信息

AFRC Institute of Animal Physiology and Genetics Research, Babraham, Cambridge.

出版信息

J Physiol. 1991 Feb;433:663-76. doi: 10.1113/jphysiol.1991.sp018449.

Abstract
  1. The whole-cell recording mode of the patch-clamp technique was used to investigate the ionic currents of enterocytes isolated from the small intestine epithelium of Necturus maculosus. 2. When enterocytes were bathed in a physiological Ringer solution and dialysed with a K(+)-rich intracellular solution containing 1.5 x 10(-7) M intracellular Ca2+, strongly outwardly rectifying currents were observed. These currents were absent from enterocytes internally dialysed with K(+)-free solutions. 3. The kinetics of activation of the outwardly rectifying current was monoexponential with rate constants decreasing with depolarization from 160 ms at 20 mV to 40 ms at 60 mV. Similar voltage dependence of the relaxations after activation were observed. 4. Strongly buffering intracellular Ca2+ with EGTA inhibited outward currents, while increasing Ca2+ increased both their magnitude and rate of activation. 5. Bath application of the K+ channel inhibitors Ba2+ and TEA greatly attenuated outwardly rectifying currents. This observation plus the fact that tail currents reverse near EK points to K+ as the charge carrier in these currents. 6. Outside-out patches showed maxi K+ channels and lower conductance K+ channels. Averaging fluctuations of the maxi K+ channels gave a kinetic behaviour similar to the whole-cell currents.
摘要
  1. 采用膜片钳技术的全细胞记录模式,研究了从斑泥螈小肠上皮分离的肠上皮细胞的离子电流。2. 当肠上皮细胞浸浴在生理林格氏溶液中,并用含有1.5×10⁻⁷M细胞内Ca²⁺的富含K⁺的细胞内溶液进行透析时,观察到强烈的外向整流电流。在用无K⁺溶液进行内部透析的肠上皮细胞中未观察到这些电流。3. 外向整流电流的激活动力学呈单指数形式,速率常数随着去极化而降低,从20mV时的160ms降至60mV时的40ms。激活后弛豫的电压依赖性也类似。4. 用乙二醇双四乙酸(EGTA)强烈缓冲细胞内Ca²⁺会抑制外向电流,而增加Ca²⁺则会增加其幅度和激活速率。5. 浴加K⁺通道抑制剂Ba²⁺和TEA可大大减弱外向整流电流。这一观察结果以及尾电流在EK附近反转的事实表明K⁺是这些电流中的电荷载体。6. 外翻膜片显示有大电导K⁺通道和低电导K⁺通道。对大电导K⁺通道的波动进行平均得到的动力学行为与全细胞电流相似。

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