Specialized cells tag sexual and species identity in Drosophila melanogaster

Abstract

Social interactions depend on individuals recognizing each other, and in this context many organisms use chemical signals to indicate species and sex1. Cuticular hydrocarbon signals are used by insects, including Drosophila melanogaster, to distinguish conspecific individuals from others1,2,3. These chemicals also contribute to intraspecific courtship and mating interactions1,2,3. However, the possibility that sex and species identification are linked by common chemical signalling mechanisms has not been formally tested. Here we provide direct evidence that a single compound is used to communicate female identity among D. melanogaster, and to define a reproductive isolation barrier between D. melanogaster and sibling species. A transgenic manipulation eliminated cuticular hydrocarbons by ablating the oenocytes, specialized cells required for the expression of these chemical signals. The resulting oenocyte-less (oe-) females elicited the normal repertoire of courtship behaviours from males, but were actually preferred over wild-type females by courting males. In addition, wild-type males attempted to copulate with oe- males. Thus, flies lacking hydrocarbons are a sexual hyperstimulus. Treatment of virgin females with the aversive male pheromone cis-vaccenyl acetate (cVA) significantly delayed mating of oe- females compared to wild-type females. This difference was eliminated when oe- females were treated with a blend of cVA and the female aphrodisiac (7Z,11Z)-heptacosadiene (7,11-HD), showing that female aphrodisiac compounds can attenuate the effects of male aversive pheromones. 7,11-HD also was shown to have a crucial role in heterospecific encounters. Specifically, the species barrier was lost because males of other Drosophila species courted oe- D. melanogaster females, and D. simulans males consistently mated with them. Treatment of oe- females with 7,11-HD restored the species barrier, showing that a single compound can confer species identity. These results identify a common mechanism for sexual and species recognition regulated by cuticular hydrocarbons.

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Figure 1: Oenocytes produce cuticular hydrocarbons.
Figure 2: Hydrocarbons regulate sexual attractiveness.
Figure 3: 7,11-HD regulates courtship between Drosophila species.
Figure 4: Mating between D. melanogaster females and D. simulans males is prevented by 7,11-HD.

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Acknowledgements

We thank H. Pavlou for help with RNA collection; N. Gompel for sharing fly strains; and A. Dornan, S. Douglas, J. C. Hall, I. Dover, C. Kent, C. Kyriacou, K. Luoto, S. Goodwin, M. Ritchie, J. Schneider and M. Sokolowski for critical comments on the manuscript. J.-C.B. was supported by a fellowship for advanced researcher from the Swiss National Science Foundation. This work was supported by the Canadian Institutes of Health Research and Canada Research Chair grants awarded to J.D.L.

Author Contributions J.-C.B. and J.D.L designed and interpreted the study. J.-C.B., J.G.M. and J.D.L. wrote the paper. J.-C. B. generated the Gal4 transgenic line. J.-C.B. and J.A. performed the behavioural experiments and GC-FID analyses, and analysed the behavioural and analytical chemical data. J.J.K. performed the anatomical analyses of the oenocyte-less flies. J.G.M. synthesized and characterized 7-T and 7,11-HD. All authors read and commented on the paper.

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Correspondence to Joel D. Levine.

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Supplementary Information

The file contains Supplementary Figures 1-3, Supplementary Tables 1-5, Supplementary Methods and additional references. (PDF 1229 kb)

Supplementary Movie

Supplementary Movie 1 shows two Oenocyte-less males engaged in head-to-head interaction. (MOV 10057 kb)

Supplementary Movie

Supplementary Movie 2 shows two Oenocyte-less males with one male attempting copulation towards the head of the other. (MOV 19817 kb)

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Billeter, J., Atallah, J., Krupp, J. et al. Specialized cells tag sexual and species identity in Drosophila melanogaster. Nature 461, 987–991 (2009). https://doi.org/10.1038/nature08495

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