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小鼠B淋巴细胞系中具有多个电导水平的阴离子通道。

Anion channels with multiple conductance levels in a mouse B lymphocyte cell line.

作者信息

Bosma M M

机构信息

Department of Physiology, University of California, Los Angeles 90024.

出版信息

J Physiol. 1989 Mar;410:67-90. doi: 10.1113/jphysiol.1989.sp017521.

Abstract
  1. Multiple conductance level ion channels were recorded in excised and cell-attached patches from cells of a mouse B lymphocyte hybridoma line. The reversal potential for the single-channel current was unaffected by the species of cation on the cytoplasmic face of the patch, but changed as the Cl- concentration was altered, indicating that the channel is anion selective. 2. The permeability sequence determined from reversal potentials was F- greater than I- greater than SCN- greater than Br- greater than Cl- greater than glucuronate greater than NO3- greater than aspartate. This was different from the conductance sequence (Cl- greater than SCN- = F- greater than Br- greater than NO3- greater than I- greater than glucuronate greater than aspartate), indicating interaction of ions within the pore of the channel. Consistent with this was the observation of anomalous mole fraction dependence with a mixed solution of thiocyanate and chloride. 3. In addition to the main open level (about 400 pS; excised patch, symmetrical 165 mM-Cl-), three subconductance levels and one supraconductance level were observed. These were concluded to be integral components of the same channel based on coincidence of appearance and identical permeabilities. 4. The channel is voltage dependent, with open probability in excised patches increasing with more positive potentials. The channel was reversibly blocked in a voltage-dependent manner by SITS (4-acetamido-4'-isothiocyanostilbene-2,2'-disulphonic acid), a stilbene derivative, on the cytoplasmic face. 5. Several differences were noted between cell-attached and excised-patch recordings. The multiple conductance level channel was less frequently seen in cell-attached patches but could often be induced to appear by prolonged application of positive voltages. This induced channel in attached patches showed an altered voltage dependence which could be partially mimicked in excised patches by including cyclic AMP and ATP in the solution on the cytoplasmic side of the membrane.
摘要
  1. 在从小鼠B淋巴细胞杂交瘤系细胞上切除的膜片和细胞贴附膜片中记录到了多电导水平离子通道。单通道电流的反转电位不受膜片胞质面阳离子种类的影响,但随着Cl⁻浓度的改变而变化,这表明该通道对阴离子具有选择性。2. 根据反转电位确定的通透序列为F⁻>I⁻>SCN⁻>Br⁻>Cl⁻>葡萄糖醛酸根>NO₃⁻>天冬氨酸。这与电导序列(Cl⁻>SCN⁻ = F⁻>Br⁻>NO₃⁻>I⁻>葡萄糖醛酸根>天冬氨酸)不同,表明离子在通道孔内存在相互作用。与此一致的是,观察到硫氰酸盐和氯化物混合溶液存在异常摩尔分数依赖性。3. 除了主要的开放水平(约400 pS;切除的膜片,对称的165 mM-Cl⁻)外,还观察到了三个亚电导水平和一个超电导水平。基于出现的一致性和相同的通透性,这些被认为是同一通道的组成部分。4. 该通道具有电压依赖性,在切除的膜片中,开放概率随着电位更正而增加。该通道在胞质面被一种芪衍生物SITS(4-乙酰氨基-4'-异硫氰酸芪-2,2'-二磺酸)以电压依赖性方式可逆性阻断。5. 在细胞贴附记录和切除膜片记录之间发现了几个差异。多电导水平通道在细胞贴附膜片中较少见,但通过长时间施加正电压常常可诱导其出现。这种在贴附膜片中诱导出的通道显示出改变的电压依赖性,在切除的膜片中,通过在膜细胞质侧的溶液中加入环磷酸腺苷和三磷酸腺苷可部分模拟这种改变。

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