Abstract
Pheromone receptors (PRs) of moths are expressed on the dendritic membrane of odorant receptor neurons (ORNs) housed in the long trichoid sensilla (TS) of antennae and are essential to sex pheromone reception. The function of peripheral neurons of Mythimna separata in recognizing sex pheromones is still unclear. In this study, electroantennogram recordings were performed from male and female antennae of M. separata, and showed that the major component of sex pheromones, (Z)-11-hexadecenal (Z11–16:Ald), evoked the strongest response of male antennae with significant differences between sexes. Single sensillum recording was used to record responses of neurons housed in TS of male M. separata. The results revealed four types of TS with three neurons housed in each type, based on profiles of responses to sex pheromone components and pheromone analogs. ORN-B of type-I TS was specifically tuned to the major sex pheromone component Z11–16:Ald; ORN-Bs in type-III and type-IV TSs were, respectively, activated by minor components (Z)-11-hexadecen-1-yl acetate (Z11–16:OAc) and hexadecenal (16:Ald); and ORNs in type-II TS were mainly activated by the sex pheromone analogs. We further cloned full-length sequences of six putative PR genes and an Orco gene. Functional characterization of PRs in the Xenopus oocyte system demonstrated that male antennae-biased MsepPR1 responded strongly to (Z)-9-tetradecenal (Z9-14:Ald), suggesting that MsepPR1 may be expressed in type-II TS. MsepPR6 was exclusively tuned to (Z)-9-tetradecen-1-yl acetate (Z9–14:OAc). MsepPR2 and MsepPR4 showed no responses to any tested components. Female antennae-biased MespPR5 was broadly tuned to Z9–14:Ald, Z9–14:OAc, Z11–16:Ald, and (Z)-11-hexadecen-1-ol (Z11–16:OH). Our results further enriched the sex pheromone recognition mechanism in the peripheral nervous system of moth M. separata.
Highlights
Pheromone-based sexual communication in moths has become an excellent model system for investigating the molecular mechanism of sensory perception because of the surprisingly high specificity in insect olfaction (Gould et al, 2010; Leal, 2013; Liu et al, 2013)
The results showed that all tested compounds elicited EAG responses of male antennae at the dose of 100 μg (Figure 1)
The responses of male antennae were significantly evoked by analog Z9–14:Ald
Summary
Pheromone-based sexual communication in moths has become an excellent model system for investigating the molecular mechanism of sensory perception because of the surprisingly high specificity in insect olfaction (Gould et al, 2010; Leal, 2013; Liu et al, 2013). Peripheral reception of pheromones in moths involves multiple proteins in male antennae, including pheromone binding proteins (PBPs), pheromone receptors (PRs), pheromone degrading enzymes, and sensory neuron membrane proteins (SNMPs; Leal, 2005; Rutzler and Zwiebel, 2005; Touhara and Vosshall, 2009). Moth sex pheromones are generally a blend of pheromone components detected by odorant receptor neurons (ORNs) housed in trichoid sensilla (TS) of male antennae (Kaissling, 1986; Heinbockel and Kaissling, 1996; Hallem et al, 2004; Hansson and Stensmyr, 2011). PRs expressed on the dendrite membrane of ORNs in the peripheral olfactory system of male antennae play a significant role in detecting conspecific sex pheromones (Vogt, 2005; Tanaka et al, 2009)
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