Noxious chemical discrimination by Tribolium castaneum TRPA1 channel in the HEK293 cell expression system

IF 2.2 Q1 ENTOMOLOGY Current Research in Insect Science Pub Date : 2023-01-01 DOI:10.1016/j.cris.2023.100066

Kenji Shimomura , Hinoki Oikawa , Kosuke Yamamoto , Takehito Terajima , Shunsuke Yajima , Motohiro Tomizawa

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Abstract

Nociception is the sensory perception of noxious chemical stimuli. Repellent behavior to avoid noxious stimuli is indispensable for survival, and this mechanism has been evolutionarily conserved across a wide range of species, from mammals to insects. The transient receptor potential ankyrin 1 (TRPA1) channel is one of the most conserved noxious chemical sensors. Here, we describe the heterologous stable expression of Tribolium castaneum TRPA1 (TcTRPA1) in human embryonic kidney (HEK293) cells. The intracellular Ca²⁺ influx was measured when two compounds, citronellal and l-menthol, derived from plant essential oils, were applied in vitro using a fluorescence assay. The analysis revealed that citronellal evoked Ca²⁺ influx dose-dependently for TcTRPA1, whereas l-menthol did not. In combination with our present and previous results of the avoidance-behavioral assay at the organism level, we suggest that TcTRPA1 discriminates between these two toxic compounds, and diversification in the chemical nociception selectivity has occurred in TRPA1 channel among insect taxa.

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在HEK293细胞表达系统中，蓖麻TRPA1通道对有害化学物质的鉴别

伤害感受是对有害化学刺激的感觉。躲避有害刺激的排斥行为对生存是必不可少的，从哺乳动物到昆虫，这种机制在进化上已经在广泛的物种中得到了保护。瞬时受体电位锚蛋白1（TRPA1）通道是最保守的有毒化学传感器之一。在这里，我们描述了谷草Tribolium castaneum TRPA1（TcTRPA1）在人类胚胎肾（HEK293）细胞中的异源稳定表达。当两种来源于植物精油的化合物香茅醛和l-薄荷醇在体外使用荧光测定法时，测量细胞内Ca2+内流。分析表明，香茅醛对TcTRPA1的Ca2+内流具有剂量依赖性，而l-薄荷醇则没有。结合我们目前和以前在生物体水平上的回避行为测定结果，我们认为TcTRPA1区分这两种有毒化合物，并且昆虫分类群中TRPA1通道的化学伤害选择性发生了多样化。

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