Tohse N, Sperelakis N
Department of Physiology and Biophysics, University of Cincinnati College of Medicine, Ohio 45267-0576.
Am J Physiol. 1990 Aug;259(2 Pt 2):H639-42. doi: 10.1152/ajpheart.1990.259.2.H639.
Single-channel currents were recorded in cultured embryonic chick (3-day-old) cardiomyocytes in cell-attached patch-clamp experiments. The patch electrode contained 50 mM Ba2+. The cell was bathed in an external solution containing 150 mM K+ (pH 7.4) at room temperature. Depolarizing pulses above -30 mV, from a holding potential of -80 mV, elicited inward unitary currents. The conductance of the channel for this unitary current was 26 pS. The activity of these channels was completely blocked by nifedipine (3 microM). These results indicate that the channel is a slow (L-type) Ca2+ channel. The channels exhibited long-lasting openings, in addition to conventional brief openings. These long openings resembled the long openings produced by the dihydropyridine Ca2+ agonist BAY K 8644 and resultant mode 2 behavior (Hess et al., Nature Lond. 311: 538-544, 1984). The long-lasting openings were observed in 25 patches out of a total of 29 patches in which single-channel activity was present. High open-state probability (Po) sweeps (with Po greater than 0.65), which mainly contain long-lasting openings, accounted for 20.7% of all sweeps. The open-time histogram for the Ca2+ channels was fitted by two exponential components. The time constants of the two components were 0.45 ms (fast) and 6.30 ms (slow). These kinetic properties were similar to those of the previous reports using BAY K 8644. Thus the slow (L-type) Ca2+ channels in young embryonic chick heart cells naturally produce many long-lasting openings in the absence of any added dihydropyridine Ca2+ agonist.
在细胞贴附式膜片钳实验中,对培养的胚胎鸡(3日龄)心肌细胞记录单通道电流。膜片电极含有50 mM Ba2+。细胞在室温下浸浴于含有150 mM K+(pH 7.4)的外部溶液中。从 -80 mV的 holding 电位施加高于 -30 mV的去极化脉冲,引发内向单位电流。该单位电流通道的电导为26 pS。这些通道的活性被硝苯地平(3 microM)完全阻断。这些结果表明该通道是一种慢(L型)Ca2+通道。除了传统的短暂开放外,这些通道还表现出持久开放。这些长开放类似于二氢吡啶Ca2+激动剂BAY K 8644产生的长开放以及由此产生的模式2行为(Hess等人,《自然》伦敦311: 538 - 544, 1984)。在总共29个存在单通道活性的膜片中,有25个膜片观察到了持久开放。主要包含持久开放的高开放态概率(Po)扫描(Po大于0.65)占所有扫描的20.7%。Ca2+通道的开放时间直方图由两个指数成分拟合。这两个成分的时间常数分别为0.45 ms(快)和6.30 ms(慢)。这些动力学特性与之前使用BAY K 8644的报道相似。因此,在未添加任何二氢吡啶Ca2+激动剂的情况下,幼龄胚胎鸡心脏细胞中的慢(L型)Ca2+通道自然会产生许多持久开放。