Specialized cells tag sexual and species identity in Drosophila melanogaster


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.


  1. 1

    Wyatt, T. D. Pheromones and Animal Behaviour: Communication by Smell and Taste (Cambridge Univ. Press, 2003)

    Google Scholar 

  2. 2

    Jallon, J. M. A few chemical words exchanged by Drosophila during courtship and mating. Behav. Genet. 14, 441–478 (1984)

    CAS  Article  Google Scholar 

  3. 3

    Ferveur, J. F. Cuticular hydrocarbons: their evolution and roles in Drosophila pheromonal communication. Behav. Genet. 35, 279–295 (2005)

    Article  Google Scholar 

  4. 4

    Coyne, J. A., Crittenden, A. P. & Mah, K. Genetics of a pheromonal difference contributing to reproductive isolation in Drosophila . Science 265, 1461–1464 (1994)

    ADS  CAS  Article  Google Scholar 

  5. 5

    Savarit, F., Sureau, G., Cobb, M. & Ferveur, J. F. Genetic elimination of known pheromones reveals the fundamental chemical bases of mating and isolation in Drosophila . Proc. Natl Acad. Sci. USA 96, 9015–9020 (1999)

    ADS  CAS  Article  Google Scholar 

  6. 6

    Grillet, M., Dartevelle, L. & Ferveur, J. F. A Drosophila male pheromone affects female sexual receptivity. Proc. Biol. Sci. 273, 315–323 (2006)

    CAS  Article  Google Scholar 

  7. 7

    Lacaille, F. et al. An inhibitory sex pheromone tastes bitter for Drosophila males. PLoS One 2, e661 (2007)

    ADS  Article  Google Scholar 

  8. 8

    Antony, C., Davis, T. L., Carlson, D. A., Pechine, J. M. & Jallon, J. M. Compared behavioral responses of male Drosophila melanogaster (Canton S) to natural and synthetic aphrodisiacs. J. Chem. Ecol. 11, 1617–1629 (1985)

    CAS  Article  Google Scholar 

  9. 9

    Ferveur, J. F. et al. Genetic feminization of pheromones and its behavioral consequences in Drosophila males. Science 276, 1555–1558 (1997)

    CAS  Article  Google Scholar 

  10. 10

    Fan, Y., Zurek, L., Dykstra, M. J. & Schal, C. Hydrocarbon synthesis by enzymatically dissociated oenocytes of the abdominal integument of the German Cockroach, Blattella germanica . Naturwissenschaften 90, 121–126 (2003)

    ADS  CAS  PubMed  Google Scholar 

  11. 11

    Marcillac, F., Bousquet, F., Alabouvette, J., Savarit, F. & Ferveur, J. F. A mutation with major effects on Drosophila melanogaster sex pheromones. Genetics 171, 1617–1628 (2005)

    CAS  Article  Google Scholar 

  12. 12

    Krupp, J. J. et al. Social experience modifies pheromone expression and mating behavior in male Drosophila melanogaster . Curr. Biol. 18, 1373–1383 (2008)

    CAS  Article  Google Scholar 

  13. 13

    Venken, K. J. & Bellen, H. Emerging technologies for gene manipulation in Drosophila melanogaster . Nature Rev. Genet. 6, 167–178 (2005)

    CAS  Article  Google Scholar 

  14. 14

    Zhou, L. et al. Cooperative functions of the reaper and head involution defective genes in the programmed cell death of Drosophila central nervous system midline cells. Proc. Natl Acad. Sci. USA 94, 5131–5136 (1997)

    ADS  CAS  Article  Google Scholar 

  15. 15

    Gutierrez, E., Wiggins, D., Fielding, B. & Gould, A. P. Specialized hepatocyte-like cells regulate Drosophila lipid metabolism. Nature 445, 275–280 (2007)

    ADS  CAS  Article  Google Scholar 

  16. 16

    McGuire, S. E., Le, P. T., Osborn, A. J., Matsumoto, K. & Davis, R. L. Spatiotemporal rescue of memory dysfunction in Drosophila . Science 302, 1765–1768 (2003)

    ADS  CAS  Article  Google Scholar 

  17. 17

    Butterworth, F. M. Lipids of Drosophila: a newly detected lipid in the male. Science 163, 1356–1357 (1969)

    ADS  CAS  Article  Google Scholar 

  18. 18

    Mane, S. D., Tompkins, L. & Richmond, R. C. Male Esterase 6 catalyzes the synthesis of a sex pheromone in Drosophila melanogaster females. Science 222, 419–421 (1983)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Kurtovic, A., Widmer, A. & Dickson, B. J. A single class of olfactory neurons mediates behavioural responses to a Drosophila sex pheromone. Nature 446, 542–546 (2007)

    ADS  CAS  Article  Google Scholar 

  20. 20

    Imhof, M., Harr, B., Brem, G. & Schlötterer, C. Multiple mating in wild Drosophila melanogaster revisited by microsatellite analysis. Mol. Ecol. 7, 915–917 (1998)

    CAS  Article  Google Scholar 

  21. 21

    Hosken, D. J., Stockley, P., Tregenza, T. & Wedell, N. Monogamy and the battle of the sexes. Annu. Rev. Entomol. 54, 361–378 (2009)

    CAS  Article  Google Scholar 

  22. 22

    Jallon, J. M. & David, J. R. Variations in cuticular hydrocarbons among the eight species of the Drosophila melanogaster subgroup. Evolution 41, 294–302 (1987)

    PubMed  Google Scholar 

  23. 23

    Cobb, M. & Jallon, J. Pheromones, mate recognition and courtship stimulation in the Drosophila melanogaster species subgroup. Anim. Behav. 39, 1058–1067 (1990)

    Article  Google Scholar 

  24. 24

    Cowling, D. E. & Burnet, B. Courtship songs and genetic control of their acoustic characteristics in sibling species of the Drosophila melanogaster subgroup. Anim. Behav. 29, 924–935 (1981)

    Article  Google Scholar 

  25. 25

    Kyriacou, C. P. & Hall, J. C. Interspecific genetic control of courtship song production and reception in Drosophila . Science 232, 494–497 (1986)

    ADS  CAS  Article  Google Scholar 

  26. 26

    Wheeler, D. A. et al. Molecular transfer of a species-specific behavior from Drosophila simulans to Drosophila melanogaster . Science 251, 1082–1085 (1991)

    ADS  CAS  Article  Google Scholar 

  27. 27

    Kent, C., Azanchi, R., Smith, B., Formosa, A. & Levine, J. D. Social context influences chemical communication in D. melanogaster males. Curr. Biol. 18, 1384–1389 (2008)

    CAS  Article  Google Scholar 

  28. 28

    Barolo, S., Carver, L. A. & Posakony, J. W. GFP and β-galactosidase transformation vectors for promoter/enhancer analysis in Drosophila . Biotechniques 29, 726–732 (2000)

    CAS  Article  Google Scholar 

  29. 29

    Russo, C. A., Takezaki, N. & Nei, M. Molecular phylogeny and divergence times of drosophilid species. Mol. Biol. Evol. 12, 391–404 (1995)

    CAS  PubMed  Google Scholar 

  30. 30

    Cobb, M., Connolly, K. & Burnet, B. Courtship behaviour in the melanogaster species sub-group of Drosophila Behaviour 95, 203–230 (1985)

    Article  Google Scholar 

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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, JC., 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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