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斜纹夜蛾 slowpoke 通道的电生理和药理学特性。

Electrophysiological and pharmacological properties of the slowpoke channel in the diamondback moth, Plutella xylostella.

机构信息

Institute of Agricultural Biotechnology, Hunan Academy of Agricultural Sciences, Changsha 410125, China; Longping Branch, College of Biology, Hunan University, Changsha 410125, China.

Institute of Agricultural Biotechnology, Hunan Academy of Agricultural Sciences, Changsha 410125, China; Hunan Provincial Key Laboratory of Pesticide Biology and Precise Use Technology, Changsha 410125, China; Key Laboratory of Pesticide Assessment, Ministry of Agriculture and Rural Affairs, Changsha 410125, China.

出版信息

Pestic Biochem Physiol. 2024 Mar;200:105824. doi: 10.1016/j.pestbp.2024.105824. Epub 2024 Feb 9.

Abstract

The slowpoke channel responds to the intracellular calcium concentration and the depolarization of the cell membrane. It plays an important role in maintaining the resting potential and regulating the homeostasis of neurons, but it can also regulate circadian rhythm, sperm capacitation, ethanol tolerance, and other physiological processes in insects. This renders it a potentially useful target for the development of pest control strategies. There are relatively few studies on the slowpoke channels in lepidopteran pests, and their pharmacological properties are still unclear. So, in this study, the slowpoke gene of Plutella xylostella (Pxslo) was heterologous expressed in HEK293T cells, and the I-V curve of the slowpoke channel was measured by whole cell patch clamp recordings. Results showed that the slowpoke channel could be activated at -20 mV with 150 μM Ca. The subsequent comparison of the electrophysiological characteristics of the alternative splicing site E and G deletions showed that the deletion of the E site enhances the response of the slowpoke channel to depolarization, while the deletion of the G site weakens the response of the slowpoke channel to depolarization. Meanwhile, the nonspecific inhibitors TEA and 4-AP of the Kv channels, and four pesticides were tested and all showed an inhibition effect on the PxSlo channel at 10 or 100 μM, suggesting that these pesticides also target the slowpoke channel. This study enriches our understanding of the slowpoke channel in Lepidopteran insects and can aid in the development of relevant pest management strategies.

摘要

缓激肽通道响应细胞内钙离子浓度和细胞膜去极化。它在维持静息电位和调节神经元的内稳态方面起着重要作用,但也可以调节昆虫的生物钟节律、精子获能、乙醇耐受性和其他生理过程。这使得它成为开发害虫防治策略的一个潜在有用的目标。在鳞翅目害虫中,关于缓激肽通道的研究相对较少,其药理学特性尚不清楚。因此,在这项研究中,我们在 HEK293T 细胞中异源表达了小菜蛾(Plutella xylostella)的缓激肽通道基因,并通过全细胞膜片钳记录测量缓激肽通道的 I-V 曲线。结果表明,缓激肽通道可以在-20 mV 时被 150 μM Ca 激活。随后对替代剪接位点 E 和 G 缺失的电生理特性进行比较,结果表明 E 位点的缺失增强了缓激肽通道对去极化的反应,而 G 位点的缺失则减弱了缓激肽通道对去极化的反应。同时,Kv 通道的非特异性抑制剂 TEA 和 4-AP,以及四种农药在 10 或 100 μM 时都对 PxSlo 通道表现出抑制作用,这表明这些农药也靶向缓激肽通道。本研究丰富了我们对鳞翅目昆虫中缓激肽通道的认识,并有助于开发相关的害虫管理策略。

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