Vaughan P, Trotter M
Can J Physiol Pharmacol. 1982 May;60(5):604-9. doi: 10.1139/y82-081.
Sartorius muscle fibres of Xenopus laevis were depolarized in solutions of high K+ content and then voltage clamped in solutions in which K+ was replaced by Rb+ and(or) tetraethylammonium ion (TEA+) at pH 9. A three-microelectrode clamp system was used in which the bath was held at virtual ground using an operational amplifier in a current meter configuration. The holding potential was set at zero membrane current potential (resting potential), close to -25 mV. A two-pulse paradigm was used to test the effects of conditioning the membrane at voltages away from the resting potential on initial currents at positive test potentials. In the absence of TEA+ rapidly rising outward currents were generated at positive test potentials, following hyperpolarizing conditioning. These currents inactivated in time and obscured predicted chloride currents. When TEA+ was added to the solution (60 mequiv./L) the currents at positive potentials rose more slowly and declined either very slowly or not at all. Projection of these current waveforms, by curve fitting, to the instant of potential change gave a sigmoid dependence of test current on conditioning voltage that was predicted from earlier results. Predictably there is a test voltage at which the initial current is independent of conditioning potential: from the data it appears that this is not necessarily the resting potential, but the cause of the shift is not clear. The results also indicated that there is a component of outward current that is very small, apparently carried by cations ("delayed rectifier current"), that does not inactivate, even at potentials more positive than the resting potential.
非洲爪蟾的缝匠肌纤维在高钾溶液中去极化,然后在pH值为9的溶液中进行电压钳制,其中钾离子被铷离子和(或)四乙铵离子(TEA⁺)取代。使用三微电极钳制系统,其中浴槽通过电流计配置中的运算放大器保持在虚拟地电位。钳制电位设置为零膜电流电位(静息电位),接近 -25 mV。使用双脉冲范式来测试在远离静息电位的电压下对膜进行预处理对正测试电位下的初始电流的影响。在没有TEA⁺的情况下,在超极化预处理后,正测试电位会产生快速上升的外向电流。这些电流会随时间失活并掩盖预测的氯离子电流。当向溶液中加入TEA⁺(60 mequiv./L)时,正电位下的电流上升得更慢,下降得非常缓慢或根本不下降。通过曲线拟合将这些电流波形投影到电位变化瞬间,得到测试电流对预处理电压的S形依赖性,这与早期结果预测的一致。可以预见,存在一个测试电压,在此电压下初始电流与预处理电位无关:从数据来看,这不一定是静息电位,但其偏移的原因尚不清楚。结果还表明,存在一个非常小的外向电流成分,显然由阳离子携带(“延迟整流电流”),即使在比静息电位更正的电位下也不会失活。