Martin R J
Br J Pharmacol. 1985 Feb;84(2):445-61. doi: 10.1111/j.1476-5381.1985.tb12929.x.
gamma-Aminobutyric acid (GABA)- and piperazine-activated single-channel currents were recorded from the bag region of the somatic muscle of the nematode parasite Ascaris suum. Cell-attached and outside-out patch-clamp techniques were used. Clean membranes were routinely prepared using collagenase. GABA (concentrations greater than 1 microM) or piperazine (concentrations greater than 200 microM) applied to the extracellular surface of the patches brought about the opening of channels producing rectangular shaped current pulses of varying duration but essentially constant amplitude. The I/V relationships of the single-channel currents for both agonists were linear and had conductances in the region of 22 pS (in symmetrical 170 mM Cl-). The reversal potential was near 0 mV when Cl- was equally distributed on both sides of the membrane. Occasionally two subconductance states were seen. The mainstate single-channel permeability was estimated to be 4 X 10(-14) cm3 s-1. At low concentrations of GABA (3-4 microM), the effective mean channel open time was in the region of 32 ms (-75 mV, 22 degrees C, cell-attached patches). At low concentrations of piperazine (500 microM) the effective mean open channel lifetime was shorter, in the region of 14 ms (-75 mV, 22 degrees C cell-attached patches). For each agonist the channel open lifetime distributions were best described by the sum of two exponentials suggesting two open mainstates. Channel openings occurred as single events and in bursts with brief closed periods within bursts. The channel closed time histograms at these concentrations were best described by the sum of up to three exponentials, suggesting the presence of three closed states. Channel open times showed no appreciable voltage sensitivity. Before desensitization, increases in agonist concentration produced an increase in the probability of the channel being open. The increased probability was associated with an increase in the frequency of channel opening, an increase in the effective mean channel open time, an increase in burst duration, an increase in the number of openings per burst, together with a reduction in the proportion of brief openings. Desensitization was seen as a decline in the probability of the channel being opened during prolonged applications of agonist. It was associated with the appearance of very long (seconds) closed periods. The distributions of the closed channel times were then best described by up to four exponentials.
从线虫寄生虫猪蛔虫体壁肌肉的袋状区域记录了γ-氨基丁酸(GABA)和哌嗪激活的单通道电流。使用了细胞贴附式和外向式膜片钳技术。常规使用胶原酶制备清洁的细胞膜。将GABA(浓度大于1μM)或哌嗪(浓度大于200μM)施加到膜片的细胞外表面会导致通道开放,产生持续时间不同但幅度基本恒定的矩形电流脉冲。两种激动剂的单通道电流的I/V关系均为线性,在对称的170 mM Cl-中电导约为22 pS。当Cl-在膜两侧均匀分布时,反转电位接近0 mV。偶尔会观察到两个亚电导状态。估计主状态单通道渗透率为4×10(-14) cm3 s-1。在低浓度GABA(3 - 4μM)时,有效平均通道开放时间约为32 ms(-75 mV,22℃,细胞贴附式膜片)。在低浓度哌嗪(500μM)时,有效平均开放通道寿命较短,约为14 ms(-75 mV,22℃,细胞贴附式膜片)。对于每种激动剂,通道开放寿命分布最好用两个指数之和来描述,表明存在两个开放主状态。通道开放以单个事件和爆发形式出现,爆发内有短暂的关闭期。这些浓度下的通道关闭时间直方图最好用多达三个指数之和来描述,表明存在三个关闭状态。通道开放时间没有明显的电压敏感性。在脱敏之前,激动剂浓度增加会导致通道开放概率增加。增加的概率与通道开放频率增加、有效平均通道开放时间增加、爆发持续时间增加、每次爆发的开放次数增加以及短暂开放比例降低有关。脱敏表现为在长时间应用激动剂期间通道开放概率下降。这与出现非常长(数秒)的关闭期有关。此时关闭通道时间的分布最好用多达四个指数来描述。