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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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.
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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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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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DOI: https://doi.org/10.1038/nn.2515
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