Brehm P, Kullberg R, Moody-Corbett F
J Physiol. 1984 May;350:631-48. doi: 10.1113/jphysiol.1984.sp015222.
Patch-clamp recordings of current through acetylcholine-activated channels were made from non-junctional membrane of innervated myotomal muscle from Xenopus laevis. Two classes of acetylcholine (ACh) receptor channels were identified on the basis of current amplitudes. Both amplitude classes exhibited current-voltage relations which deviated from linearity as the extrapolated reversal potential was approached (-5 to -12 mV). Over the range of greatest linearity the conductances of the two classes were 64 and 44 pS. Both event classes were blocked by alpha-bungarotoxin. At the normal resting membrane potential (approximately -95 mV) the larger conductance channel (gamma) exhibited an apparent mean channel open time of less than 1 ms, compared to approximately 2 ms for the smaller gamma class. The apparent open time was voltage-dependent, changing e-fold with a 63 mV hyperpolarization for the high gamma channel and 93 mV hyperpolarization for the low gamma channel. At low ACh concentrations (0.1-0.3 microM) both amplitude classes exhibited bursts of successive openings separated by brief closures of less than 0.5 ms. Bursts were separated by longer closed intervals of 1 to greater than 100 ms. Closed interval histograms revealed corresponding populations of brief and long closures, indicating that at least two kinetic processes are required to describe the distribution of closed intervals. In the absence of exogenous ACh, channels were observed in an occasional patch which showed a conductance and extrapolated reversal potential similar to ACh-activated channels. In such patches the event frequency could occasionally be altered by adjusting the negative pressure applied to the patch. The two main conductance classes of ACh activated channels were observed to coexist in most patches. However, the most frequent event observed in non-junctional membrane of innervated muscle corresponded to the high gamma class. In this respect, the non-junctional ACh receptors bore a greater similarity to junctional ACh receptors than to non-junctional receptors reported for denervated muscle.
利用膜片钳技术记录了非洲爪蟾受神经支配的肌节肌肉非连接膜上乙酰胆碱激活通道的电流。根据电流幅度鉴定出两类乙酰胆碱(ACh)受体通道。两类通道的电流 - 电压关系在接近外推反转电位(-5至-12 mV)时均偏离线性。在最大线性范围内,两类通道的电导分别为64和44 pS。两类事件均被α-银环蛇毒素阻断。在正常静息膜电位(约-95 mV)下,较大电导通道(γ)的表观平均通道开放时间小于1 ms,而较小的γ类通道约为2 ms。表观开放时间与电压有关,高γ通道超极化63 mV、低γ通道超极化93 mV时开放时间呈指数变化。在低ACh浓度(0.1 - 0.3 μM)下,两类幅度的通道均表现出连续开放的簇状活动,中间间隔短暂关闭,关闭时间小于0.5 ms。簇状活动之间被1至大于100 ms的较长关闭间隔分开。关闭间隔直方图显示了相应的短暂和长时间关闭群体,表明至少需要两个动力学过程来描述关闭间隔的分布。在没有外源性ACh的情况下,偶尔在膜片中观察到的通道显示出与ACh激活通道相似的电导和外推反转电位。在这种膜片中,事件频率偶尔可通过调整施加在膜片上的负压来改变。在大多数膜片中观察到ACh激活通道的两种主要电导类共存。然而,在受神经支配肌肉的非连接膜中观察到的最常见事件对应于高γ类。在这方面,非连接ACh受体与连接ACh受体的相似性大于去神经肌肉报道的非连接受体。