Department of Neurobiology, Harvard Medical School, Boston, Massachusetts 02115.
Department of Neurobiology, Harvard Medical School, Boston, Massachusetts 02115
J Neurosci. 2014 Jul 9;34(28):9182-9. doi: 10.1523/JNEUROSCI.0339-14.2014.
SNX-482, a peptide toxin isolated from tarantula venom, has become widely used as an inhibitor of Cav2.3 voltage-gated calcium channels. Unexpectedly, we found that SNX-482 dramatically reduced the A-type potassium current in acutely dissociated dopamine neurons from mouse substantia nigra pars compacta. The inhibition persisted when calcium was replaced by cobalt, showing that it was not secondary to a reduction of calcium influx. Currents from cloned Kv4.3 channels expressed in HEK-293 cells were inhibited by SNX-482 with an IC50 of <3 nM, revealing substantially greater potency than for SNX-482 inhibition of Cav2.3 channels (IC50 20-60 nM). At sub-saturating concentrations, SNX-482 produced a depolarizing shift in the voltage dependence of activation of Kv4.3 channels and slowed activation kinetics. Similar effects were seen on gating of cloned Kv4.2 channels, but the inhibition was less pronounced and required higher toxin concentrations. These results reveal SNX-482 as the most potent inhibitor of Kv4.3 channels yet identified. Because of the effects on both Kv4.3 and Kv4.2 channels, caution is needed when interpreting the effects of SNX-482 on cells and circuits where these channels are present.
从狼蛛毒液中分离出的肽毒素 SNX-482 已被广泛用作 Cav2.3 电压门控钙通道的抑制剂。出乎意料的是,我们发现 SNX-482 可显著降低从小鼠黑质致密部急性分离的多巴胺神经元中的 A 型钾电流。当用钴取代钙时,抑制作用仍然存在,表明这不是钙内流减少的结果。在表达于 HEK-293 细胞的克隆 Kv4.3 通道中的电流被 SNX-482 抑制,IC50<3 nM,表明其对 SNX-482 抑制 Cav2.3 通道的作用(IC50 为 20-60 nM)具有显著更高的效力。在亚饱和浓度下,SNX-482 产生 Kv4.3 通道激活的电压依赖性去极化偏移,并减慢激活动力学。在克隆 Kv4.2 通道的门控中也观察到类似的作用,但抑制作用不那么明显,需要更高的毒素浓度。这些结果表明 SNX-482 是迄今为止鉴定出的最有效的 Kv4.3 通道抑制剂。由于对 Kv4.3 和 Kv4.2 通道的影响,当在存在这些通道的细胞和电路中解释 SNX-482 的作用时需要谨慎。