Volatile allosteric antagonists of mosquito odorant receptors inhibit human-host attraction.

Odorant-dependent behaviors in insects are triggered by the binding of odorant ligands to the variable subunits of heteromeric olfactory receptors. Previous studies have shown, however, that specific odor binding to ORco, the common subunit of odorant receptor heteromers, may allosterically alter olfactory receptor function and profoundly affect subsequent behavioral responses. Using an insect cell-based screening platform, we identified and characterized several antagonists of the odorant receptor coreceptor of the African malaria vector Anopheles gambiae (AgamORco) in a small collection of natural volatile organic compounds. Because some of the identified antagonists were previously shown to strongly repel Anopheles and Culex mosquitoes, we examined the bioactivities of the identified antagonists against Aedes, the third major genus of the Culicidae family. The tested antagonists inhibited the function of Ae. aegypti ORco ex vivo and repelled adult Asian tiger mosquitoes (Ae. albopictus). Binary mixtures of specific antagonists elicited higher repellency than single antagonists, and binding competition assays suggested that this enhanced repellence is due to antagonist interaction with distinct ORco sites. Our results also suggest that the enhanced mosquito repellency by antagonist mixtures is due to additive rather than synergistic effects of the specific antagonist combinations on ORco function. Taken together, these findings provide novel insights concerning the molecular aspects of odorant receptor function. Moreover, our results demonstrate that a simple screening assay may be used for the identification of allosteric modifiers of olfactory-driven behaviors capable of providing enhanced personal protection against multiple mosquito-borne infectious diseases.

Identification and functional characterization of olfactory indolergic receptors in Drosophila melanogaster.

Indole-sensitive odorant receptors or indolORs belong to a mosquito-specific expansion as ancient as the Culicidae lineage. Brachyceran flies appeared to lack representative members of this group despite the importance of indolics in this important group of dipterans. To explore whether indolORs occur in other brachyceran species, we searched for candidate indolORs in Drosophila melanogaster. Using phylogenetic tools, we show that D. melanogaster OR30a, OR43a, and OR49b form a distinct monophyletic lineage with mosquito indolORs. To explore a potential functional orthology with indolORs, we expressed these three Drosophila ORs in Xenopus laevis oocytes and measured their responses to a panel of indolic compounds. We provide evidence that OR30a, OR43a, and OR49b exhibit high sensitivity to indoles. Along with the recent discovery of indolORs in the housefly Musca domestica, our findings suggest that indolORs are a widespread feature of the peripheral olfactory systems of Diptera.

Effects of fluctuating thermal regimes on cold survival and life history traits of the spotted wing Drosophila (Drosophila suzukii).

Drosophila suzukii is an invasive pest causing severe damages to a large panel of cultivated crops. To facilitate its biocontrol with strategies such as sterile or incompatible insect techniques, D. suzukii must be mass-produced and then stored and transported under low temperature. Prolonged cold exposure induces chill injuries that can be mitigated if the cold period is interrupted with short warming intervals, referred to as fluctuating thermal regimes (FTR). In this study, we tested how to optimally use FTR to extend the shelf life of D. suzukii under cold storage. Several FTR parameters were assessed: temperature (15, 20, 25 °C), duration (0.5, 1, 2, 3 h), and frequency (every 12, 24, 36, 48 h) of warming intervals, in two wild-type lines and in two developmental stages (pupae and adults). Generally, FTR improved cold storage tolerance with respect to constant low temperatures (CLT). Cold mortality was lower when recovery temperature was 20 °C or higher, when duration was 2 h per day or longer, and when warming interruptions occurred frequently (every 12 or 24 h). Applying an optimized FTR protocol to adults greatly reduced cold mortality over long-term storage (up to 130 d). Consequences of FTR on fitness-related traits were also investigated. For adults, poststorage survival was unaffected by FTR, as was the case for female fecundity and male mating capacity. On the other hand, when cold storage occurred at pupal stage, poststorage survival and male mating capacity were altered under CLT, but not under FTR. After storage of pupae, female fecundity was lower under FTR compared to CLT, suggesting an energy trade-off between repair of chill damages and egg production. This study provides detailed information on the application and optimization of an FTR-based protocol for cold storage of D. suzukii that could be useful for the biocontrol of this pest.

Corrigendum: Supersensitive Odorant Receptor Underscores Pleiotropic Roles of Indoles in Mosquito Ecology.

[This corrects the article DOI: 10.3389/fncel.2018.00533.].

Supersensitive Odorant Receptor Underscores Pleiotropic Roles of Indoles in Mosquito Ecology.

Mosquitoes exhibit highly diverse and fast evolving odorant receptors (ORs). The indole-sensitive OR gene clade, comprised of Or2 and Or10 is a notable exception on account of its conservation in both mosquito subfamilies. This group of paralogous genes exhibits a complex developmental expression pattern in Aedes aegypti: AaegOr2 is expressed in both adults and larvae, AaegOr10 is adult-specific and a third member named AaegOr9 is larva-specific. OR2 and OR10 have been deorphanized and are selectively activated by indole and skatole, respectively. Using the two-electrode voltage clamp of Xenopus oocytes expressing Ae. aegypti ORs, we show that AaegOR9 is supersensitive and narrowly tuned to skatole. Our findings suggest that Ae. aegypti has evolved two distinct molecular strategies to detect skatole in aquatic and terrestrial environments, highlighting the central ecological roles of indolic compounds in the evolutionary and life histories of these insects.