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Control of sexual differentiation and behavior by the doublesex gene in Drosophila melanogaster

Nature Neuroscience volume 13, pages 458–466 (2010)Cite this article

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Abstract

Doublesex proteins, which are part of the structurally and functionally conserved Dmrt gene family, are important for sex determination throughout the animal kingdom. We inserted Gal4 into the doublesex (dsx) locus of Drosophila melanogaster, allowing us to visualize and manipulate cells expressing dsx in various tissues. In the nervous system, we detected differences between the sexes in dsx-positive neuronal numbers, axonal projections and synaptic density. We found that dsx was required for the development of male-specific neurons that coexpressed fruitless (fru), a regulator of male sexual behavior. We propose that dsx and fru act together to form the neuronal framework necessary for male sexual behavior. We found that disrupting dsx neuronal function had profound effects on male sexual behavior. Furthermore, our results suggest that dsx-positive neurons are involved in pre- to post-copulatory female reproductive behaviors.

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Figure 1: Expression of GAL4 from the dsxGal4 locus recapitulates endogenous dsx expression.
Figure 2: Sexually dimorphic expression of dsxGal4-neurons and associated projections in 5-d adult CNS.
Figure 3: Sex-specific dsxGal4 expression in the foreleg and effect of basitarsal amputations on axonal projections.
Figure 4: Colocalization of FruM neuronal cells and dsxGal4-expressing cells (expressing UAS-nGFP) in 3-d-old adult male flies.
Figure 5: dsxGal4 expression in CNS of FruM-null males and females expressing FruM, DsxM or the anti-apoptotic transgene p35.
Figure 6: dsxGal4 neurons control male sexual behavior.
Figure 7: dsxGal4 neurons control female sexual behavior.

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Acknowledgements

We thank T. Aigaki, P. Breslin, B. Dickson, J.-F. Ferveur, F. Hirth, G. Lee, R. Renkawitz-Pohl, Y. Rong and S. Sweeney for providing valuable reagents and protocols, and J.-C. Billeter, J. Hall, J. Levine, B. Oliver and S. Waddell for comments on the manuscript. We also thank members of the Goodwin laboratory for helpful discussions. This work was supported by grants from the Wellcome Trust.

Author information

Author notes

  1. Elizabeth J Rideout

    Present address: Present address: Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada.,

  2. Elizabeth J Rideout, Anthony J Dornan and Megan C Neville: These authors contributed equally to this work.

Authors and Affiliations

  1. Faculty of Biomedical and Life Sciences, Integrative and Systems Biology, University of Glasgow, Glasgow, UK

    Elizabeth J Rideout, Anthony J Dornan, Megan C Neville, Suzanne Eadie & Stephen F Goodwin

  2. Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK

    Megan C Neville & Stephen F Goodwin

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  1. Elizabeth J Rideout
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Contributions

E.J.R., A.J.D., M.C.N. and S.F.G. designed experiments and wrote the paper. E.J.R., A.J.D. and M.C.N. all contributed equally to performing the experiments. S.E. provided technical assistance.

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Correspondence to Stephen F Goodwin.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6 and Supplementary Tables 1 and 2 (PDF 11079 kb)

Supplementary Video 1

Sexually dimorphic dsxGAL4 neural circuitry in 5 d adult brains. Movies generated from confocal optical slice stacks, moving ventrally to dorsally (anterior top), of dsxGAL4 driving UAS-mCD8::GFP (membrane-bound GFP) male (top) and female (bottom) whole sample brains. Presenting comparative representations of the sexually dimorphic axonal projection patterns associated with the dsxGAL4-expressing neuronal cell bodies with respect to surrounding anatomical landmarks. These stacks were used to generate the maximal Z projections represented in figures 2b and 2e. Neuronal projections expressing UAS-mCD8::GFP (membrane-bound GFP) stained with anti-mCD8 antibody (green). Neuropil counterstained with anti-nC82 Mab (magenta). (MOV 3821 kb)

Supplementary Video 2

Sexually dimorphic dsxGAL4 neural circuitry in 5 day adult ventral nerve cords. Movies generated from confocal optical slice stacks, moving ventrally to dorsally (anterior top), of dsxGAL4 driving UAS-mCD8::GFP (membrane-bound GFP) male (left) and female (right) whole sample ventral nerve cords. Presenting comparative representations of the sexually dimorphic axonal projection patterns associated with the dsxGAL4-expressing neuronal cell bodies with respect to surrounding anatomical landmarks. These stacks were used to generate the maximal Z projections represented in figures 2i and 2k. Neuronal projections expressing UAS-mCD8::GFP (membrane-bound GFP) stained with anti-mCD8 antibody (green). Neuropil counterstained with anti-nC82 Mab (magenta). (MOV 3663 kb)

Supplementary Video 3

Wild-type male and UAS-TNTG;dsxGAL4 female. A wild-type male is shown courting and then copulating with a UAS-TNTG;dsxGAL4 female. (MOV 15634 kb)

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Rideout, E., Dornan, A., Neville, M. et al. Control of sexual differentiation and behavior by the doublesex gene in Drosophila melanogaster. Nat Neurosci 13, 458–466 (2010). https://doi.org/10.1038/nn.2515

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