Effects of RDL GABA Receptor Point Mutants on Susceptibility to Meta-Diamide and Isoxazoline Insecticides in Drosophila melanogaster.

Simple Summary: Insects have special receptors called RDL that regulate the effects of a neurotransmitter GABA in their nervous system. Interestingly, these receptors are also the main targets of many insecticides used to control pests. We used fruit flies with different variants of the Rdl gene and tested four new types of meta-diamide and isoxazoline insecticides. The results showed that some mutations made the flies less sensitive to certain insecticides, while another mutation made the flies super resistant to all of the insecticides tested. Using computer simulations, we explored how these mutations might be affecting the insecticides' ability to attach. We found that specific areas between parts of the receptor seem to be important for these new insecticides to work. This helps us understand how insects can resist these insecticides and might lead to better ways to control pests in the future. Ionotropic γ-aminobutyric acid (GABA) receptors in insects, specifically those composed of the RDL (resistant to dieldrin) subunit, serve as important targets for commonly used synthetic insecticides. These insecticides belong to various chemical classes, such as phenylpyrazoles, cyclodienes, meta-diamides, and isoxazolines, with the latter two potentially binding to the transmembrane inter-subunit pocket. However, the specific amino acid residues that contribute to the high sensitivity of insect RDL receptors to these novel insecticides remain elusive. In this study, we investigated the susceptibility of seven distinct Drosophila melanogaster Rdl point mutants against four meta-diamide and isoxazoline insecticides: isocycloseram, fluxametamide, fluralaner, and broflanilide. Our findings indicate that, despite exhibiting increased sensitivity to fluralaner in vitro, the RdlI276C mutant showed resistance to isocycloseram and fluxametamide. Similarly, the double-points mutant RdlI276F+G279S also showed decreased sensitivity to the tested isoxazolines. On the other hand, the RdlG335M mutant displayed high levels of resistance to all tested insecticides. Molecular modeling and docking simulations further supported these findings, highlighting similar binding poses for these insecticides. In summary, our research provides robust in vivo evidence supporting the idea that the inter-subunit amino acids within transmembrane M1 and M3 domains form the binding site crucial for meta-diamide and isoxazoline insecticide interactions. This study highlights the complex interplay between mutations and insecticide susceptibility, paving the way for more targeted pest control strategies. [ABSTRACT FROM AUTHOR]