Lotshaw David P
Department of Biological Sciences, Northern Illinois University, DeKalb, 60115, USA.
J Membr Biol. 2006 Mar;210(1):51-70. doi: 10.1007/s00232-005-7012-x. Epub 2006 Jun 22.
Multiple genes of the TASK subfamily of two-pore domain K(+) channels are reported to be expressed in rat glomerulosa cells. To determine which TASK isoforms contribute to native leak channels controlling resting membrane potential, patch-clamp studies were performed to identify biophysical and pharmacological characteristics of macroscopic and unitary K(+) currents diagnostic of recombinant TASK channel isoforms. Results indicate K(+) conductance (gK(+)) is mediated almost exclusively by a weakly voltage-dependent (leak) K(+) channel closely resembling TASK-3. Leak channels exhibited a unitary conductance approximating that expected for TASK-3 under the recording conditions employed, brief mean open times and a voltage-dependent open probability. Extracellular H(+) induced voltage-independent inhibition of gK(+), exhibiting an IC(50) of 56 nM: (pH 7.25) and a Hill coefficient of 0.75. Protons inhibited leak channel open probability (P(o)) by promoting a long-lived closed state (tau > 500 ms). Extracellular Zn(2+) mimicked the effects of H(+); inhibition of gK(+) exhibited an IC(50) of 41 microM: with a Hill coefficient of 1.26, inhibiting channel gating by promoting a long-lived closed state. Ruthenium red (5 microM: ) inhibited gK(+) by 75.6% at 0 mV. Extracellular Mg(2+) induced voltage-dependent block of gK(+), inhibiting unitary current amplitude without affecting mean open time. Bupivacaine induced voltage-dependent block of gK(+), exhibiting IC(50) values of 116 microM: at -100 mV and 28 microM: at 40 mV with Hill coefficients of 1 at both potentials. Halothane induced a voltage-independent stimulation of gK(+) primarily by decreasing the leak channel closed-state dwell time.
据报道,双孔域钾离子通道(K(+)通道)的TASK亚家族的多个基因在大鼠肾小球细胞中表达。为了确定哪些TASK亚型对控制静息膜电位的天然渗漏通道有贡献,进行了膜片钳研究,以鉴定重组TASK通道亚型的宏观和单通道钾离子电流的生物物理和药理学特征。结果表明,钾离子电导(gK(+))几乎完全由一种与TASK-3非常相似的弱电压依赖性(渗漏)钾离子通道介导。渗漏通道的单通道电导在所用记录条件下接近TASK-3预期值,平均开放时间短暂且开放概率呈电压依赖性。细胞外氢离子(H(+))引起gK(+)的电压非依赖性抑制,半数抑制浓度(IC(50))为56 nM(pH 7.25),希尔系数为0.75。质子通过促进长寿命关闭状态(τ>500毫秒)来抑制渗漏通道的开放概率(P(o))。细胞外锌离子(Zn(2+))模拟了氢离子的作用;对gK(+)的抑制表现出IC(50)为41 μM,希尔系数为1.26,通过促进长寿命关闭状态来抑制通道门控。钌红(5 μM)在0 mV时使gK(+)抑制75.6%。细胞外镁离子(Mg(2+))引起gK(+)的电压依赖性阻断,抑制单通道电流幅度而不影响平均开放时间。布比卡因引起gK(+)的电压依赖性阻断,在-100 mV时IC(50)值为116 μM,在40 mV时为28 μM,在两个电位下希尔系数均为1。氟烷主要通过减少渗漏通道关闭状态的停留时间引起gK(+)的电压非依赖性刺激。