Department of Molecular and Integrative Physiology, University of Michigan Medical School, Ann Arbor, MI 48109-5622, USA.
Am J Physiol Heart Circ Physiol. 2011 Nov;301(5):H1984-95. doi: 10.1152/ajpheart.00304.2011. Epub 2011 Sep 2.
Cardiac ventricular myocytes possess an extensive t-tubular system that facilitates the propagation of membrane potential across the cell body. It is well established that ionic currents at the restricted t-tubular space may lead to significant changes in ion concentrations, which, in turn, may affect t-tubular membrane potential. In this study, we used the whole cell patch-clamp technique to study accumulation and depletion of t-tubular potassium by measuring inward rectifier potassium tail currents (I(K1,tail)), and inward rectifier potassium current (I(K1)) "inactivation". At room temperatures and in the absence of Mg(2+) ions in pipette solution, the amplitude of I(K1,tail) measured ~10 min after the establishment of whole cell configuration was reduced by ~18%, but declined nearly twofold in the presence of 1 mM cyanide. At ~35°C I(K1,tail) was essentially preserved in intact cells, but its amplitude declined by ~85% within 5 min of cell dialysis, even in the absence of cyanide. Intracellular Mg(2+) ions played protective role at all temperatures. Decline of I(K1,tail) was accompanied by characteristic changes in its kinetics, as well as by changes in the kinetics of I(K1) inactivation, a marker of depletion of t-tubular K(+). The data point to remodeling of t tubules as the primary reason for the observed effects. Consistent with this, detubulation of myocytes using formamide-induced osmotic stress significantly reduced I(K1,tail), as well as the inactivation of inward I(K1). Overall, the data provide strong evidence that changes in t tubule volume/structure may occur on a short time scale in response to various types of stress.
心肌细胞具有广泛的 T 管系统,有助于跨细胞体传播膜电位。离子电流在受限的 T 管空间中可能导致离子浓度发生显著变化,这反过来又可能影响 T 管膜电位,这一点已得到充分证实。在这项研究中,我们使用全细胞膜片钳技术通过测量内向整流钾尾电流(I(K1,tail))和内向整流钾电流(I(K1))“失活”来研究 T 管钾的积累和耗竭。在室温下,在没有 Mg(2+)离子的情况下,在建立全细胞构型约 10 分钟后测量到的 I(K1,tail)幅度减少了约 18%,但在存在 1mM 氰化物的情况下,其幅度下降了近两倍。在~35°C 时,完整细胞中 I(K1,tail)基本保持不变,但在细胞透析后 5 分钟内,其幅度下降了约 85%,即使没有氰化物也是如此。细胞内 Mg(2+)离子在所有温度下都起到保护作用。I(K1,tail)的下降伴随着其动力学的特征变化,以及 I(K1)失活动力学的变化,这是 T 管钾耗竭的标志。这些数据表明 T 管重塑是观察到的效应的主要原因。与这一观点一致的是,使用甲酰胺诱导的渗透压力使肌细胞去 T 管化显著降低了 I(K1,tail)以及内向 I(K1)的失活。总体而言,这些数据提供了有力的证据,表明 T 管体积/结构的变化可能在短时间内发生,以应对各种类型的应激。