Labarca P, Zapata O, Beltran C, Darszon A
Instituto de Biotechnologia, Universidad Nacional Autonoma de Mexico.
Zygote. 1995 Aug;3(3):199-206. doi: 10.1017/s0967199400002598.
Fusion of purified mouse sperm plasma membranes to planar lipid bilayers resulted in the insertion of three ion channel types. They could be discerned on the basis of their selectivity, conductance, gating and voltage-dependent properties. The presence of a previously reported large, Ca2+ -selective channel was confirmed. Here, it is reported that the Ca2+ -selective channel from mouse sperm plasma membrane displayed a PNa+/PK+ = 1.6 +/- 0.2 (n = 4) and was blocked by micromolar concentrations of ruthenium red. Fusion yielded also a cation-selective channel (PNa+/PK+ = 2.5 +/- 0.3, n = 3) with a main open conductance substate of 103 pS and a smaller open substate of 51 pS (600 mM K+ cis/100 mM Na+ trans). The channel inserted into bilayers in two stable fashions: a high-activity mode (open probability = 0.57 +/- 0.02, n = 3), and a low activity mode (open probability < 1%, n = 4). In high mode, the channel displayed bursting kinetics and burst length was voltage independent. In addition, a perfectly anion-selective channel, with a slope conductance of 83 pS (600 KCl cis/100 KCl trans), was identified. It displayed a high, nearly constant open probability (approximately 0.90) in the 0 to -80 mV range.
纯化的小鼠精子质膜与平面脂质双层融合后,导致三种离子通道类型的插入。根据它们的选择性、电导率、门控和电压依赖性特性可以区分这些通道。先前报道的一种大的、Ca2+ 选择性通道的存在得到了证实。在此报道,来自小鼠精子质膜的Ca2+ 选择性通道表现出PNa+/PK+ = 1.6 +/- 0.2(n = 4),并被微摩尔浓度的钌红阻断。融合还产生了一种阳离子选择性通道(PNa+/PK+ = 2.5 +/- 0.3,n = 3),其主要开放电导亚态为103 pS,较小的开放亚态为51 pS(600 mM K+ 顺式/100 mM Na+ 反式)。该通道以两种稳定方式插入双层膜中:一种高活性模式(开放概率 = 0.57 +/- 0.02,n = 3)和一种低活性模式(开放概率 < 1%,n = 4)。在高模式下,该通道表现出爆发动力学,爆发长度与电压无关。此外,还鉴定出一种完美的阴离子选择性通道,其斜率电导为83 pS(600 KCl顺式/100 KCl反式)。在0至 -80 mV范围内,它表现出高的、几乎恒定的开放概率(约0.90)。