Oliveira Eugênio E, Du Yuzhe, Nomura Yoshiko, Dong Ke
Department of Entomology, Genetics and Neuroscience Programs, Michigan State University, East Lansing, MI 48824, USA.
Neurotoxicology. 2013 Sep;38:42-50. doi: 10.1016/j.neuro.2013.06.001. Epub 2013 Jun 10.
Voltage-gated sodium channels are critical for electrical signaling in the nervous system. Pyrethroid insecticides exert their toxic action by modifying the gating of sodium channels. A valine to methionine mutation in the transmembrane segment 6 of domain I (IS6) of sodium channels from tobacco budworms (Heliothis virescens) has been shown to alter channel gating and reduce insect sodium channel sensitivity to pyrethroids. A valine to leucine substitution was subsequently reported in pyrethroid-resistant bedbug populations. Intriguingly, pyrethroid-resistant mammalian sodium channels possess an isoleucine at the corresponding position. To determine whether different substitutions at this position alter channel gating and confer pyrethroid resistance, we made valine to methionine, isoleucine or leucine substitutions at the corresponding position, V409, in a cockroach sodium channel and examined the gating properties and pyrethroid sensitivity of the three mutants in Xenopus oocytes. All three mutations reduced the channel sensitivity to three pyrethroids (permethrin, cismethrin and deltamethrin). V409M, but not V409I or V409L, caused 6-7mV depolarizing shifts in the voltage dependences of both activation and inactivation. V409M and V409L slowed channel activation kinetics and accelerated open-state deactivation kinetics, but V409I did not. Furthermore, the substitution of isoleucine with valine, but not with methionine nor leucine, at the corresponding position in a rat skeletal muscle sodium channel, rNav1.4, enhanced channel sensitivity to deltamethrin. Collectively, our study highlights an important role of residues at 409 in regulating not only sodium channel gating, but also the differential sensitivities of insect and mammalian sodium channels to pyrethroids.
电压门控钠通道对神经系统中的电信号传导至关重要。拟除虫菊酯类杀虫剂通过改变钠通道的门控来发挥其毒性作用。烟草夜蛾(烟芽夜蛾)钠通道结构域I的跨膜片段6(IS6)中的缬氨酸到甲硫氨酸突变已被证明会改变通道门控并降低昆虫钠通道对拟除虫菊酯的敏感性。随后在抗拟除虫菊酯的臭虫种群中报道了缬氨酸到亮氨酸的替换。有趣的是,抗拟除虫菊酯的哺乳动物钠通道在相应位置具有异亮氨酸。为了确定该位置的不同替换是否会改变通道门控并赋予拟除虫菊酯抗性,我们在蟑螂钠通道的相应位置V409处进行了缬氨酸到甲硫氨酸、异亮氨酸或亮氨酸的替换,并在非洲爪蟾卵母细胞中检测了这三个突变体的门控特性和拟除虫菊酯敏感性。所有这三个突变都降低了通道对三种拟除虫菊酯(氯菊酯、顺式氯氰菊酯和溴氰菊酯)的敏感性。V409M,但不是V409I或V409L,在激活和失活的电压依赖性上引起了6 - 7mV的去极化偏移。V409M和V409L减慢了通道激活动力学并加速了开放状态失活动力学,但V409I没有。此外,在大鼠骨骼肌钠通道rNav1.4的相应位置用缬氨酸替换异亮氨酸,而不是用甲硫氨酸或亮氨酸,增强了通道对溴氰菊酯的敏感性。总的来说,我们的研究强调了409位残基不仅在调节钠通道门控方面,而且在昆虫和哺乳动物钠通道对拟除虫菊酯的不同敏感性方面都起着重要作用。