Brinkmeier H, Zachar E, Rüdel R
Abteilung für Allgemeine Physiologie, Universität Ulm, Federal Republic of Germany.
Pflugers Arch. 1991 Nov;419(5):486-91. doi: 10.1007/BF00370793.
The voltage-dependent K+ channels of the mammalian sarcolemma were studied with the patch-clamp technique in intact, enzymatically dissociated fibres from the toe muscle of the mouse. With a physiological solution (containing 2.5 mM K+) in the pipette, depolarizing pulses imposed on a cell-attached membrane patch activated K+ channels with a conductance of about 17 pS. No channel activity was observed when the pipette solution contained 2 mM tetraethylammonium (TEA), or 2 mM 4-aminopyridine (4-AP). Whole cell recordings from these very small muscle fibres showed the well-known delayed rectifier K+ outward current with a threshold of about -40 mV. The whole-cell current was completely blocked by 2 mM TEA in the bath, suggesting that the TEA-sensitive channels in the patch were also delayed rectifier channels. The inactivation properties of the channels were studied in the cell-attached mode. Averaged single-channel traces showed at least two types of channels discernible by their inactivation time course at a test potential of 60 mV. The fast type inactivated with a time constant of about 150 ms, the slow type with a time constant of about 400 ms. A little channel activity always remained during pulses lasting several minutes, indicating either the presence of a very slowly inactivating third type of K+ channel, or the tendency of the fast inactivating channels to re-open at constant voltage. No difference was seen in the single-channel amplitudes of the different types of K+ channels. The well characterized adenosine-5'-triphosphate-(ATP)-sensitive and Ca(2+)-dependent K+ channels, although present, were not active under the conditions used.(ABSTRACT TRUNCATED AT 250 WORDS)
利用膜片钳技术,在完整的、酶解分离的小鼠趾肌纤维中研究了哺乳动物肌膜上的电压依赖性钾通道。当移液管中加入生理溶液(含2.5 mM钾离子)时,施加于细胞贴附膜片上的去极化脉冲可激活电导约为17 pS的钾通道。当移液管溶液中含有2 mM四乙铵(TEA)或2 mM 4-氨基吡啶(4-AP)时,未观察到通道活性。对这些非常小的肌纤维进行全细胞记录,显示出众所周知的延迟整流钾外向电流,阈值约为 -40 mV。浴液中加入2 mM TEA可完全阻断全细胞电流,这表明膜片中对TEA敏感的通道也是延迟整流通道。在细胞贴附模式下研究了通道的失活特性。在60 mV的测试电位下,平均单通道记录显示至少有两种类型的通道,可根据其失活时间进程区分。快速型通道的失活时间常数约为150 ms,慢速型通道的失活时间常数约为400 ms。在持续数分钟的脉冲期间,总会有少量通道活性存在,这表明要么存在一种失活非常缓慢的第三种钾通道类型,要么快速失活通道有在恒定电压下重新开放的趋势。不同类型钾通道的单通道幅度未见差异。尽管存在特征明确的腺苷-5'-三磷酸(ATP)敏感性和钙依赖性钾通道,但在所使用的条件下它们并不活跃。(摘要截短于250字)