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Dominant-negative subunits reveal potassium channel families that contribute to M-like potassium currents.

机译:显性负亚基揭示了钾通道家族,这些通道家族有助于M样钾电流。

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M-currents are K+ currents generated by members of the KCNQ family of K+ channels (Wang et al., 1998). However, in some cells, M-like currents may be contaminated by members of other K+ channel gene families, such as the erg family (Meves et al., 1999; Selyanko et al., 1999). In the present experiments, we have used the acute expression of pore-defective mutants of KCNQ3 (DN-KCNQ3) and Merg1a (DN-Merg1a) as dominant negatives to separate the contributions of these two families to M-like currents in NG108-15 neuroblastoma hybrid cells and rat sympathetic neurons. Two kinetically and pharmacologically separable components of M-like current could be recorded from NG108-15 cells that were individually suppressed by DN-Merg1a and DN-KCNQ3, respectively. In contrast, only DN-KCNQ3, and not DN-Merg1a, reduced currents recorded from sympathetic neurons. Pharmacological tests suggested that the residual current in DN-KCNQ3-treated sympathetic neurons was carried by residual KCNQ channels. Ineffectiveness of DN-Merg1a in sympathetic neurons was not caused by lack of expression, as judged by confocal microscopy of Flag-tagged DN-Merg1a. These results accord with previous inferences regarding the roles of erg and KCNQ channels in generating M-like currents. This experimental approach should therefore be useful in delineating the contributions of members of these two gene families to K+ currents in other cells.
机译:M电流是由K +通道的KCNQ家族成员生成的K +电流(Wang等,1998)。但是,在某些细胞中,M样电流可能会被其他K +通道基因家族(例如erg家族)的成员污染(Meves等,1999; Selyanko等,1999)。在本实验中,我们使用了KCNQ3(DN-KCNQ3)和Merg1a(DN-Merg1a)的孔缺陷突变体的急性表达作为显性阴性,以分离这两个家族对NG108-15中M样电流的贡献神经母细胞瘤杂交细胞和大鼠交感神经元。可以从分别被DN-Merg1a和DN-KCNQ3分别抑制的NG108-15细胞中记录M样电流的两个动力学和药理上可分离的成分。相比之下,只有DN-KCNQ3减少了从交感神经元记录的电流,而没有DN-Merg1a减少了。药理学测试表明,DN-KCNQ3处理的交感神经元中的残留电流是由残留的KCNQ通道携带的。 DN-Merg1a在交感神经元中的无效性不是由于缺乏表达所致,这是通过共聚焦显微镜对带有Flag标签的DN-Merg1a进行判断的。这些结果与先前关于erg和KCNQ通道在产生类M电流中的作用的推论一致。因此,这种实验方法在描述这两个基因家族的成员对其他细胞中K +电流的贡献时应该是有用的。

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