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Odorant-Degrading Enzymes (ODEs) are supposed to be involved in the signal inactivation step within the olfactory sensilla of insects by quickly removing odorant molecules from the vicinity of the olfactory receptors. Only three ODEs have been both identified at the molecular level and functionally characterized: two were specialized in the degradation of pheromone compounds and the last one was shown to degrade a plant odorant. Previous work has shown that the antennae of the cotton leafworm Spodoptera littoralis, a worldwide pest of agricultural crops, express numerous candidate ODEs. We focused on an esterase overexpressed in males antennae, namely SlCXE7. We studied its expression patterns and tested its catalytic properties towards three odorants, i.e. the two female sex pheromone components and a green leaf volatile emitted by host plants. SlCXE7 expression was concomitant during development with male responsiveness to odorants and during adult scotophase with the period of male most active sexual behaviour. Furthermore, SlCXE7 transcription could be induced by male exposure to the main pheromone component, suggesting a role of Pheromone-Degrading Enzyme. Interestingly, recombinant SlCXE7 was able to efficiently hydrolyze the pheromone compounds but also the plant volatile, with a higher affinity for the pheromone than for the plant compound. In male antennae, SlCXE7 expression was associated with both long and short sensilla, tuned to sex pheromones or plant odours, respectively. Our results thus suggested that a same ODE could have a dual function depending of it sensillar localisation. Within the pheromone-sensitive sensilla, SlCXE7 may play a role in pheromone signal termination and in reduction of odorant background noise, whereas it could be involved in plant odorant inactivation within the short sensilla.

Background Insects respond to the spatial and temporal dynamics of a pheromone plume, which implies not only a strong response to 'odor on', but also to 'odor off'. This requires mechanisms geared toward a fast signal termination. Several mechanisms may contribute to signal termination, among which odorant-degrading enzymes. These enzymes putatively play a role in signal dynamics by a rapid inactivation of odorants in the vicinity of the sensory receptors, although direct in vivo experimental evidences are lacking. Here we verified the role of an extracellular carboxylesterase, esterase-6 (Est-6), in the sensory physiological and behavioral dynamics of Drosophila melanogaster response to its pheromone, cis -vaccenyl acetate (cVA). Est-6 was previously linked to post-mating effects in the reproductive system of females. As Est-6 is also known to hydrolyze cVA in vitro and is expressed in the main olfactory organ, the antenna, we tested here its role in olfaction as a putative odorant-degrading enzyme. Results We first confirm that Est-6 is highly expressed in olfactory sensilla, including cVA-sensitive sensilla, and we show that expression is likely associated with non-neuronal cells. Our electrophysiological approaches show that the dynamics of olfactory receptor neuron (ORN) responses is strongly influenced by Est-6, as in Est-6° null mutants (lacking the Est-6 gene) cVA-sensitive ORN showed increased firing rate and prolonged activity in response to cVA. Est-6° mutant males had a lower threshold of behavioral response to cVA, as revealed by the analysis of two cVA-induced behaviors. In particular, mutant males exhibited a strong decrease of male-male courtship, in association with a delay in courtship initiation. Conclusions Our study presents evidence that Est-6 plays a role in the physiological and behavioral dynamics of sex pheromone response in Drosophila males and supports a role of Est-6 as an odorant-degrading enzyme (ODE) in male antennae. Our results also expand the role of Est-6 in Drosophila biology, from reproduction to olfaction, and highlight the role of ODEs in insect olfaction.

Castniid moths (Lepidoptera: Castniidae) display a butterfly-like reproductive behavior, i.e., they use visual stimuli for mate location and females have apparently lost their pheromone glands in an evolutionary context. In this paper we report for the first time the identification of three new compounds, namely n-octadecyl acetate, (Z)-9-octadecenyl acetate and (E,Z)-2,13-octadecadienyl acetate, in males of the Castniid Palm Borer, Paysandisia archon, which could be involved in its short-range courtship behavior, and also shed light on recent controversies on the sexual behavior of the species. The compounds are produced in a ring-shaped gland of the male terminalia and have occasionally been detected in very minor amounts (ng) in ovipositor extracts of females, but only while mating or just after copulation. We also report that males use the already known (E,Z)-2,13-octadecadienol to mark their territory by rubbing their midlegs against the upper side of nearby leaves, especially palm leaves. This compound, produced in large amounts, is mostly concentrated in the midleg basitarsi and its maximum production is detected on the sexually mature 1-day-old specimens. In addition, analysis of male wings extracts confirms the presence of Z,E and E,E-farnesals, which are mostly produced in the median band of hindwings of 48-53 h-old insects. The biological significance of farnesals in this species is unknown. Our results point out that the chemical communication of P. archon relies mostly on males, which appear to bear all chemical burden in this respect.

Del extracto en diclorometano de las glándulas de veneno de las hormigas cortadoras: Atta laevigata, Atta bisphaerica, Acromyrmex subterraneus molestans. Fueron analizados por CG y CG-EM e identificados el 4-metilpirrol-2-carboxilato de metilo y el 3-etil-2,5-dimetilpirazina como los compuestos principales de la feromona de trilla (marcación de camino).