Genetic characterization of ABC pesticide transporters establishes Drosophila as an in vivo model for toxicogenetics

Pesticides remain one of the most effective ways of controlling agricultural and public health insects, but much is still unknown regarding how these compounds reach their targets. Specifically, the role of ABC transporters in pesticide absorption and excretion is poorly understood, especially compared to the detailed knowledge in mammalian systems. Here, we present a comprehensive characterization of pesticide transporters in the model insect D. melanogaster . An RNAi screen was performed, which knocked down individual ABCs in targeted epithelial tissue, examining the subsequent changes in sensitivity to the pesticides spinosad and fipronil. This simultaneously implicated a novel ABC drug transporter CG4562 but also highlighted a predominant role for the P-glycoprotein orthologue Mdr65 . Further characterization of the P-glycoprotein family was performed via transgenic overexpression and immunolocalization, finding that Mdr49 and Mdr50 play enigmatic roles in pesticide toxicology perhaps determined by their different subcellular localizations within the midgut. Lastly, heterologous expression of the Mdr65 orthologue from the major malaria vector Anopheles gambiae was used to establish an in vivo characterization system for the characterization of P-glycoproteins from non-model insects in D. melanogaster . This study provides the basis for establishing Drosophila as a model for toxicology research regarding drug transporters. ### Competing Interest Statement I have read the journal's policy and the authors of this manuscript have the following competing interests: SD, NHBL recieved funding from collaboration between Bayer Crop Sciences (https://www.bayer.com/en/agriculture) and the Institute of Molecular Biology and Biotechnology (Greece). KV, RN, and SV are employees at Bayer Crop Sciences.