Singh S, Wu C F
Department of Biology, University of Iowa, Iowa City 52242.
J Exp Biol. 1990 Sep;152:59-76. doi: 10.1242/jeb.152.1.59.
The larval muscle fibers of Drosophila show four outward K+ currents in addition to the inward Ca2+ current in voltage-clamp recordings. The Shaker (Sh) and the slowpoke (slo) mutations, respectively, eliminate the voltage-activated fast K+ current (IA) and the Ca2(+)-activated fast K+ current (ICF). Quinidine specifically blocks the voltage-activated delayed K+ current (IK) at micromolar concentrations. We used Sh, slo and quinidine to remove specifically one or more K+ currents, so as to study physiological properties of these currents not previously characterized, and to examine their role in membrane excitability. A linear relationship was observed between the peak ICF and the peak ICa at different membrane potentials. ICF inactivated considerably during a 140 ms pulse to +20 mV. Recovery from inactivation was not complete for up to 2 s at the holding potential of -50 mV, which is much slower than the recovery of Ca2+ current from inactivation. In addition to IA and ICF, two delayed K+ currents are also observed in these fibers, the voltage-activated IK and the Ca2(+)-activated ICS. Near the end of a 500 ms depolarizing pulse, both IA and ICF are inactivated. Ca2(+)-free and 20 mmol l-1 Ca2+ saline were used to examine the tail currents of the remaining IK and ICS. The tail currents of ICS were slower than those of IK and reversed between -30 and -50 mV in different fibers. We further studied the dose-dependence of the blockade of IK by quinidine, which did not indicate a simple one-to-one binding mechanism. Current-clamp recordings from normal, Sh, slo and the double-mutant Sh;slo fibers suggested that ICF plays a stronger role than IA in repolarization of the larval muscle membrane. Elimination of ICF facilitates the occurrence of action potentials. Further elimination of IK prolonged the action potentials to several hundred milliseconds.
在电压钳记录中,果蝇幼虫肌纤维除了内向Ca²⁺电流外,还显示出四种外向K⁺电流。分别是Shaker(Sh)和slowpoke(slo)突变,消除了电压激活的快速K⁺电流(IA)和Ca²⁺激活的快速K⁺电流(ICF)。奎尼丁在微摩尔浓度下特异性阻断电压激活的延迟K⁺电流(IK)。我们使用Sh、slo和奎尼丁特异性去除一种或多种K⁺电流,以研究这些先前未表征的电流的生理特性,并研究它们在膜兴奋性中的作用。在不同膜电位下,观察到ICF峰值与ICa峰值之间存在线性关系。在+20 mV的140 ms脉冲期间,ICF显著失活。在-50 mV的保持电位下,失活恢复在长达2 s内不完全,这比Ca²⁺电流从失活中恢复要慢得多。除了IA和ICF外,在这些纤维中还观察到两种延迟K⁺电流,电压激活的IK和Ca²⁺激活的ICS。在500 ms去极化脉冲接近结束时,IA和ICF都失活。使用无Ca²⁺和20 mmol·l⁻¹ Ca²⁺盐水来检查剩余IK和ICS的尾电流。ICS的尾电流比IK的尾电流慢,并且在不同纤维中在-30至-50 mV之间反转。我们进一步研究了奎尼丁对IK阻断的剂量依赖性,这并未表明存在简单的一对一结合机制。来自正常、Sh、slo和双突变Sh;slo纤维的电流钳记录表明,ICF在幼虫肌膜复极化中比IA发挥更强的作用。消除ICF促进动作电位的发生。进一步消除IK将动作电位延长至数百毫秒。