Molecular characterization and functional expression of voltage-gated sodium channel variants in Apolygus lucorum...

Apolygus lucorum (Meyer-Dür) is a serious worldwide agricultural pest, especially for Bt cotton in China. Pyrethroids, neonicotinoids and organophosphates are the most effective insecticides to control piercing and sucking insects, including A. lucorum. The voltage-gated sodium channel (Na_v ) is major target site of pyrethroids. Extensive alternative splicing and RNA editing, two major post-transcriptional mechanisms, contribute to generate different functional sodium channel variants. In our research, we characterized the sodium channel variants of A. lucorum.

In this study, we isolated numerous sodium channel variants that cover the entire coding region of the VGSC gene from A. lucorum. All clones could be grouped into 47 splice types based on the presence of nine alternative exons (exons j, n, o, a, p, b, s, q and t). Exons j, b and t were located independently, while exons n, o, a and p were located adjacently, as were exons s and q. We also found 35 nucleotide changes in different positions in individual variants, of which 18 nucleotide changes were A-to-I RNA editing, 11 nucleotide changes were likely due to U-to-C or C-to-U editing, and the others were likely natural sequence polymorphisms in the population. Furthermore, we expressed all of the variants in Xenopus oocytes. Eighteen of them were expressed in oocytes and sensitive to tetrodotoxin.

Our results provide a functional basis for understanding how A. lucorum sodium channel variants work in regulating channel expression, pharmacology and gating properties for agricultural insects. Apolygus lucorum is widely distributed in cotton production. Our results suggest how AlNa_v (the sodium channel of A.lucorum) variants work in regulating channel expression, pharmacology and sodium channel gating for agricultural insects in the future. © 2020 Society of Chemical Industry.