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Neural circuit mechanisms of sexual receptivity in Drosophila females


Choosing a mate is one of the most consequential decisions a female will make during her lifetime. A female fly signals her willingness to mate by opening her vaginal plates, allowing a courting male to copulate1,2. Vaginal plate opening (VPO) occurs in response to the male courtship song and is dependent on the mating status of the female. How these exteroceptive (song) and interoceptive (mating status) inputs are integrated to regulate VPO remains unknown. Here we characterize the neural circuitry that implements mating decisions in the brain of female Drosophila melanogaster. We show that VPO is controlled by a pair of female-specific descending neurons (vpoDNs). The vpoDNs receive excitatory input from auditory neurons (vpoENs), which are tuned to specific features of the D. melanogaster song, and from pC1 neurons, which encode the mating status of the female3,4. The song responses of vpoDNs, but not vpoENs, are attenuated upon mating, accounting for the reduced receptivity of mated females. This modulation is mediated by pC1 neurons. The vpoDNs thus directly integrate the external and internal signals that control the mating decisions of Drosophila females.

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Fig. 1: Female-specific vpoDNs control vaginal plate opening (VPO).
Fig. 2: Auditory inputs to vpoDNs.
Fig. 3: vpoENs and vpoDNs are tuned to conspecific courtship song.
Fig. 4: vpoDNs integrate mating status and song.

Data availability

Confocal images of the central nervous systems of split-GAL4 lines used in this study are available at Other datasets generated during the current study are available from the corresponding author on reasonable request.


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We thank the Janelia FlyLight, Fly Facility, Project Technical Resources, Molecular Biology, Functional Connectome, and Experimental Technology teams and N. Chen for technical assistance; E. Lillvis, E. Behrman and D. Stern for providing courtship songs of various Drosophila species; and K. Svoboda, K. Feng and K. Keleman for comments on the manuscript. This work was funded by the Howard Hughes Medical Institute.

Author information




B.J.D., K.W. and F.W. conceived the study and wrote the manuscript. K.W. and F.W. performed all experiments and analysed the data. N.F., T.Y., C.P. and F.W. reconstructed selected neurons and synapses in the FAFB EM volume. R.P. managed the tracing team.

Corresponding author

Correspondence to Barry J. Dickson.

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Competing interests

The authors declare no competing interests.

Additional information

Peer review information Nature thanks the anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data figures and tables

Extended Data Fig. 1 Genetic screen for neuronal activities required for female receptivity.

a, Difference in the copulation frequencies of split-GAL4 UAS-TNTe (tetanus toxin) and control UAS-TNTe virgin females within a 10-min observation period, relative to the control group. b, Stable-split lines (SS ID) and sample size (n) for the 234 split-GAL4 lines shown in a.

Extended Data Fig. 2 Anatomical characterization of vpoDNs.

a, Confocal images of brains and ventral nerve cords from female and male flies carrying vpoDN-SS1, vpoDN-SS2, or vpoDN-SS3, and UAS-myrFLAG, stained with anti-FLAG to reveal membranes of targeted neurons (green) and mAb nc82 to reveal all synapses (magenta). One pair of vpoDNs are labelled in females but not in males. Scale bar: 100 μm. b, Confocal images of female brains showing the co-labelling of vpoDNs with dsx-LexA but not fru-LexA. Scale bar: 20 μm. c, Confocal images of female brains showing the expression of ChAT, GAD1, and vGluT in vpoDN (labelled by Halo tag, arrows), as revealed by FISH. Scale bar: 10 μm. Representative images are shown from at least 5 independent samples examined in each case.

Extended Data Fig. 3 Functional characterization of vpoDNs.

a, b, Percentage of pairs copulating (a) and frequency of VPO (b) during 10 min of courtship between a virgin female of the indicated genotype and a wild-type male. c, Percentage of time wild-type males chased or extended their wings towards the virgin female during a 10-min observation period. d, Snapshots of female VPO induced upon photoactivation of vpoDNs (Supplementary Videos 23). e, Percentage of isolated virgins performing VPO upon photoactivation of vpoDNs (5 s, 635 nm, 57 μW mm−2). Each female was tested three times as follows: first, while intact, then with the cuticle over the posterior part of ventral nerve cord removed to expose the abdominal ganglion (sham), and finally, after the abdominal nerve trunk was severed (cut). Data in b and c shown as scatter plots with mean ± s.e.m. P values in italics, two-sided Fisher’s exact test in a, two-sided Wilcoxon test in b and c. See Supplementary Table 3 for details of statistical analyses.

Extended Data Fig. 4 vpoDNs are sensitive to courtship song and are postsynaptic to auditory neurons.

a, Frequency of male courtship behaviours around the onset of female VPO. n = 124 VPOs from 12 pairs of flies. b, GCaMP6s signal changes in vpoDNs of virgins with intact aristae in response to courtship song and white noise. Lines connect data points from the same fly. Error bars show mean ± s.e.m. P values in italics, paired two-sided Wilcoxon test. See also Supplementary Table 3. c, Neurons presynaptic to a single vpoDN in the FAFB EM volume, showing the number of input synapses identified (thresholded at 10).

Extended Data Fig. 5 Split-GAL4 driver lines targeting vpoENs and vpoINs and neurotransmitter types revealed by FISH.

a, Confocal images of female central nervous system carrying indicated split-GAL4 driver lines and UAS-myrFLAG or UAS-Chrimson-mVenus. Samples were stained with anti-FLAG or anti-GFP (green) to reveal membranes of targeted neurons and mAb nc82 to reveal all synapses (magenta). Arrows indicate soma. Scale bar: 100 μm. b, Confocal images showing the expression of ChAT, GAD1, and vGluT in vpoEN and vpoIN neurons (labelled by Halo tag, arrows) in female brains, as revealed by FISH. Representative images are shown from at least 5 independent samples examined in each case.

Extended Data Fig. 6 Responses of vpoDN, vpoEN, and vpoIN towards natural and synthetic courtship songs.

a, c, e, Traces of natural and artificial songs used as auditory stimuli. b, d, f, Sound-evoked GCaMP6s responses in vpoDNs, vpoENs and vpoINs of virgin melanogaster females. Darker traces indicate mean response; grey shading indicates s.e.m. Grey bars indicate stimuli (5 s). ‘1’ and ‘2’ in a and b indicate different audio clips from the same species. Selected data for Drosophila elegans are reproduced in Fig. 3. Sample sizes were as indicated, except for responses to simulans songs, for which n = 7, 6 and 6 for vpoDN, vpoEN, and vpoIN, respectively.

Extended Data Table 1 vpoDN inputs identified by EM reconstruction
Extended Data Table 2 Synaptic connections identified by EM reconstruction

Supplementary information

Supplementary Information

This file contains Supplementary Notes, Supplementary Figure 1, Supplementary Tables 1-5 and Supplementary References.

Reporting Summary

Supplementary Video 1

| Vaginal plate opening A virgin female fly performs VPO while being courted by a male, shown at half speed (15 fps) and replayed once. The full arena is shown on the left; a close-up of the female's abdomen on the right.

Supplementary Video 2

| Optogenetic activation of vpoDNs elicits vaginal plate opening A montage of video clips of 4 virgin vpoDN-SS1 UAS-Chrimson females upon 3 s of continuous 625 nm illumination at 200 µW/mm2), shown at normal speed (30 fps).

Supplementary Video 3

| Comparison of vaginal plate opening and ovipositor extrusion Video clips of vaginal plate opening by vpoDN-SS1 UAS-Chrimson virgins (left) and ovipositor extrusion31 by DNp13-SS1 UAS-Chrimson virgins (right), induced by photoactivation with 5 s of continuous 635 nm illumination at 57 µW/mm2). Shown at normal speed (30 fps) in lateral (top) and ventral (bottom) views.

Supplementary Video 4

| vpoDN reconstructed in female brain EM volume A single vpoDN cells (red) reconstructed in the right hemisphere of the FAFB EM volume.

Supplementary Video 5

| pC1a, vpoENs and vpoINs are presynaptic to vpoDN Single pC1a (orange), vpoEN (blue) and vpoIN (purple) cells partially reconstructed in the right hemisphere of the FAFB EM volume.

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Wang, K., Wang, F., Forknall, N. et al. Neural circuit mechanisms of sexual receptivity in Drosophila females. Nature 589, 577–581 (2021).

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