Female contact modulates male aggression via a sexually dimorphic GABAergic circuit in Drosophila

Journal name:
Nature Neuroscience
Volume:
17,
Pages:
81–88
Year published:
DOI:
doi:10.1038/nn.3581
Received
Accepted
Published online

Abstract

Intraspecific male-male aggression, which is important for sexual selection, is regulated by environment, experience and internal states through largely undefined molecular and cellular mechanisms. To understand the basic neural pathway underlying the modulation of this innate behavior, we established a behavioral assay in Drosophila melanogaster and investigated the relationship between sexual experience and aggression. In the presence of mating partners, adult male flies exhibited elevated levels of aggression, which was largely suppressed by prior exposure to females via a sexually dimorphic neural mechanism. The suppression involved the ability of male flies to detect females by contact chemosensation through the pheromone-sensing ion channel ppk29 and was mediated by male-specific GABAergic neurons acting on the GABAA receptor RDL in target cells. Silencing or activating this circuit led to dis-inhibition or elimination of sex-related aggression, respectively. We propose that the GABAergic inhibition represents a critical cellular mechanism that enables prior experience to modulate aggression.

At a glance

Figures

  1. Prior female experience inhibits sex-related male-male aggression.
    Figure 1: Prior female experience inhibits sex-related male-male aggression.

    (a) Scheme for the aggression assay, four conditions are shown with color coding as naive male only (blue), experienced male only (purple), naive male + virgin female (red), and experienced male + virgin female (green). (b) Timeline of the average aggression duration of wild-type Canton-S males within 2 h. Each data point represents average aggression duration quantified in 5-min intervals. The lack of aggression in the first 30 min reflects courtship behavior (n = 4 pairs for each condition). (c) Average aggression duration of Canton-S and w1118 males under different conditions. Both genotypes showed female experience–induced inhibition of aggression in pair- or single-housed settings (n = 11 and 6 for Canton-S and w1118 in pair-housed, 8 and 7 in single-housed). Aggression interactions between 60–90 min after the onset were quantified (***P < 0.0001, *P = 0.019, **P = 0.0084, #P = 0.0059, by Student's t test). (d) The inhibition index, defined as (AggressionN – AggressionE)/AggressionN, for pair- and single-housed settings (n = 11 and 8 for pair-housed and single-housed, respectively). P = 0.1989 by Student's t test. Error bars denote s.e.m.

  2. Prior female contact-dependent inhibition of aggression is long term and requires chemosensation through the pheromone-sensing channel ppk29.
    Figure 2: Prior female contact–dependent inhibition of aggression is long term and requires chemosensation through the pheromone-sensing channel ppk29.

    (a) The inhibitory effect of prior female experience on aggression lasted up to 2 d. The experienced (E) male flies were housed with females for 24 h and were then separated from the females and reared for 0, 2, 3 or 5 d prior to the behavior assay (n = 11, 6, 7 and 5 flies for each manipulation). (b) Memory mutants showed no defects in female experience–induced inhibition. Inhibition index was comparable among Canton-S, w1118, rut, amn and Orb2−/− flies (n = 11, 6, 4, 4 and 4 flies for each genotype). (c) Inhibition index for various conditioning assays (n = 11, 6, 5, 6, 5, 5 and 7 pairs of flies for each group). D.P., exposed to D. pseudoobscura females; D.V., exposed to D. virilis females; E24h, exposed to females for 24 h; E10h, exposed to females for 10 h; Emesh, conditioned with females, but separated by a nylon mesh; Emated, exposed to mated females; Emale, exposed to extra males. (d) Inhibition of aggression by female experience was impaired specifically in mutants lacking ppk29. Compared with genotypes with defects in pheromone sensation, vision or olfaction, only ppk29−/−, ppk29-Gal4>Kir and ppk29-Gal4,fruFLP, UAS>stop>TNTact flies showed a significant reduction of inhibition index (n = 5, 4, 4, 4, 6, 5, 5 and 5 pairs of flies for each genotype). TNTina and TNTact denote the inactive and active forms of TNT, respectively. **P < 0.01, ***P < 0.001 by one-way ANOVA followed by Bonferroni's multiple comparison test (ad), and Student's t test (d, TNTina and TNTact). Error bars denote s.e.m.

  3. fru+ d5-HT1B+ sexually dimorphic neurons mediate female experience-dependent inhibition of aggression.
    Figure 3: fru+ d5-HT1B+ sexually dimorphic neurons mediate female experience–dependent inhibition of aggression.

    (a) Temperature shift–induced silencing of d5-HT1B+ neurons resulted in dis-inhibition of aggression without affecting baseline aggression (n = 5, 5 and 6 pairs of flies for each genotype). ***P < 0.0001 by one-way ANOVA followed by Bonferroni's multiple comparison test (top) or Student's t test (bottom). N, naive; E, experienced. (b) fru+ d5-HT1B+ neurons in the adult brain labeled by mCD8double colonGFP and Dscam17.1double colonGFP, including γ-neurons of the mushroom bodies and a cluster with soma located between antennal lobes and the SOG region. The sexual dimorphism was evident with more cells and more elaborated arbors being present in male brains compared with female brains. nc82 co-staining (red) showed the neuropil. Scale bars represent 40 μm. (c) Blocking chemical transmission with a TNT transgene or electrically silencing with a Kir2.1 transgene expression in fru+ d5-HT1B+ neurons led to strong dis-inhibition of aggression (n = 6, 6, 5 and 7 pairs of flies for each genotype). *P = 0.0012, **P = 0.0099, by Student's t test. (d) Inducible activation of fru+ d5-HT1B+ neurons by the heat-activated channel dTrpA1 led to reduced baseline aggression at 29 °C as compared with the 20 °C control. This reduction of baseline aggression persisted when the mushroom body neurons were excluded using MB-Gal80. The genotype for the manipulation is w+; UAS>stop>dTrpA1; fruFLP/d5-HT1B-Gal4, without or with MB-Gal80 (n = 5 pairs of flies for each genotype). ***P < 0.0001, **P = 0.0049, by Student's t test. Error bars denote s.e.m.

  4. Ectopic activation of the serotoninergic circuit elevates baseline aggression without affecting female contact-dependent inhibition of aggression.
    Figure 4: Ectopic activation of the serotoninergic circuit elevates baseline aggression without affecting female contact–dependent inhibition of aggression.

    (a,b) 5-HTP treatment elevated baseline sex-related aggression (a) without affecting female contact–dependent inhibition of aggression (b) (n = 11 and 5 pairs of flies for each condition). *P = 0.0424 by Student's t test. Error bars denote s.e.m.

  5. Female experience acts through GABAergic neurotransmission in fru+, GABA+ and d5-HT1B+ neurons to inhibit aggression.
    Figure 5: Female experience acts through GABAergic neurotransmission in fru+, GABA+ and d5-HT1B+ neurons to inhibit aggression.

    (a) fru+ GABA+ neurons displayed sexual dimorphism in the adult brain, especially in the region above the SOG, with a difference in both the cell number and the projection pattern in female and male brains. nc82 co-staining (red) showed the neuropil. Scale bar represents 40 μm. (b) A subpopulation of the fru+ d5-HT1B+ neurons (green) above the SOG region was labeled with antibody to GAD (red), indicated by arrows, suggesting that these neurons are GABAergic. The top three panels are representative max projection images and the bottom panel is an image from a single confocal optical section. Scale bars represent 10 μm. (c) Blocking neurotransmission by TNT in GABA+ fru+ neurons led to dis-inhibition of aggression. Although inhibition of fru+ d5-HT1B+ neurons by TNT resulted in dis-inhibition (Fig. 3c), adding Gad-Gal80 abolished this effect, suggesting that the GABAergic fru+ d5-HT1B+ neurons are responsible for the dis-inhibition of aggression (n = 5 pairs of flies for each genotype). (d) Activation of GABA+ neurons or GABA+ fru+ neurons by dTrpA1 at 29 °C suppressed the baseline aggression. Although activation of fru+ d5-HT1B+ neurons by dTrpA1 suppressed the baseline aggression (Fig. 3d), adding Gad-Gal80 abolished this reduction, suggesting that the GABAergic fru+ d5-HT1B+ neurons are responsible for the reduction of baseline aggression. (n = 5, 5, 5, 6, 7 and 4 pairs of flies for each genotype). ***P < 0.001 by Student's t test. Error bars denote s.e.m.

  6. GABA neurotransmission, the GABAA receptor RDL and Rdl+ fru+ sexually dimorphic neurons mediate female experience dependent inhibition of aggression.
    Figure 6: GABA neurotransmission, the GABAA receptor RDL and Rdl+ fru+ sexually dimorphic neurons mediate female experience dependent inhibition of aggression.

    (a) Blocking GABA neurotransmission in d5-HT1B+ neurons using RNAi against GAD or VGAT led to dis-inhibition of aggression (n = 5, 4, 5 and 4 pairs of flies for each genotype). *P = 0.0359, **P = 0.0139 by Student's t test. Error bars denote s.e.m. (b) Inhibition of aggression was reduced in flies with Rdl knockdown pan-neuronally (Elav-Gal4>UAS-Rdl RNAi) or with the hypomorphic allele RdlMB08800. In addition, activation of fru+ Rdl+ neurons by dTrpA1 at 29 °C resulted in dis-inhibition of male-male aggression induced by female contact (n = 4, 4, 5, 5, 8, and 6 pairs of flies for groups from left to right). *P < 0.05, **P < 0.01, ***P < 0.001, by One-way ANOVA followed by Bonferroni's multiple comparison test and Student's t test (for Rdl-Gal4,fruFLP). Error bars denote s.e.m. (c) Neurons labeled by Rdl-Gal4 were widely distributed in the brain. Scale bar represents 40 μm. (d) Sexual dimorphisms of fru+ Rdl+ neurons in the fly brain. Representative images of the male and female brains are shown. nc82 co-staining (red) showed the neuropil. Scale bars represent 40 μm.

  7. Experience with females inhibits sex-related male-male aggression.
    Supplementary Fig. 1: Experience with females inhibits sex-related male-male aggression.

    (a) Average aggression duration of cs males was quantified in 5 min intervals for 2 hours for naïve male only (blue), experienced males only (purple), naïve males + virgin females (red), experienced males + virgin females (yellow), naïve males + mated females (green), experienced males + mated females (brown). (n = 4 pairs for each condition). (b) Courtship index measured at 70–75 min for naïve male + virgin female and naive male + mated female. The latter showed significantly higher courtship index. (n = 4 pairs for each condition). (c) Average aggression frequency of cs and w1118 males was quantified for the 60-90 min. No obvious difference in the inhibition of aggression is apparent between the two genotypes. (n = 11 and 6 pairs for cs and w1118). **: P < 0.01, ***: P < 0.001 by Student's t test. Error bars denote s.e.m.

  8. Experimental schemes for behavioral analysis.
    Supplementary Fig. 2: Experimental schemes for behavioral analysis.

    (a) Detailed experimental scheme for analyzing sex-related male-male aggression and its regulation by prior female experience. (b) The dis-inhibition phenotype was examined after temperature shift to induce silencing of specific group of neurons with enhancer-Gal4 driven UAS-Gal80ts and UAS-Kir2.1.

  9. Conditioning parameters for the prior female experience dependent inhibition of aggression.
    Supplementary Fig. 3: Conditioning parameters for the prior female experience dependent inhibition of aggression.

    Inhibition of aggression were displayed by males conditioned with virgin females for 24 hours, with mated females, with virgin females that express membrane bound sex peptide using fru-Gal4, which would therefore reject males, and with Drosophila pseudoobscura. However, conditioning with virgin females for 10 hours, removing females right after mating during conditioning, conditioning males in vials previously housed virgin females, conditioning males with virgin females that were separated by a nylon mesh, conditioning with Drosophila virilis, or with extra males all failed to elicit inhibition of aggression. On the other hand, removing females after mating during testing abolished aggression, revealing the presence of females during the aggression assay is crucial. (n = 11, 11, 5, 6, 6, 6, 6, 5, 5, 5, 6 and 8 pairs of flies for each condition). ***: P < 0.001 by One-way ANOVA followed by Bonferroni's multiple comparison test. Error bars denote s.e.m.

  10. Previous female contact does not significantly affect the courtship index and locomotion of experienced males. Social hierarchy established between males housed in pairs has little to do with the female-contacts dependent inhibition of sex-related male aggression.
    Supplementary Fig. 4: Previous female contact does not significantly affect the courtship index and locomotion of experienced males. Social hierarchy established between males housed in pairs has little to do with the female-contacts dependent inhibition of sex-related male aggression.

    (a) The courtship index and climbing index of experienced vs. naive male flies did not exhibit significant differences. (n = 5 flies for courtship index and 3 groups for climbing index). (b) Two male flies that were housed in pairs but not with each other (mixed group), so that they had social experiences but no established hierarchy with each other, exhibited as much contact-dependent inhibition of the sex-related aggression as did males that were raised together (pair). (n = 5 pairs of flies for each condition). **: P = 0.0012, by Student's t-test. . Error bars denote s.e.m.

  11. Aggression duration measurements corresponding to Fig. 2a, 2b, 2d (n = 11, 6, 7 and 5 pairs of flies for each condition).
    Supplementary Fig. 5: Aggression duration measurements corresponding to Fig. 2a, 2b, 2d (n = 11, 6, 7 and 5 pairs of flies for each condition).

    ***: P < 0.001, by One-way ANOVA followed by Bonferroni's multiple comparison test.. Error bars denote s.e.m.

  12. Sexual dimorphic distribution of fru+, d5-HT1B+ neurons in the brain and VNC of adult male and female flies.
    Supplementary Fig. 6: Sexual dimorphic distribution of fru+, d5-HT1B+ neurons in the brain and VNC of adult male and female flies.

    (a) fru+, d5-HT1B+ neurons in the adult male brain labeled by mCD8::GFP including γ-neurons of the mushroom bodies and a cluster with soma located below the antennal lobes and above the SOG region. Scale bar = 40 μm. The magnified view is shown in the lower left panel. Scale bar = 10 μm. nc82 co-staining in red shows the neuropil. A 3D rendering of the neuropil is shown in the lower right panel. (b) A subpopulation of fru+, d5-HT1B+ neurons independent of mushroom bodies are required for the prior female-contacts dependent inhibition of aggression. MB-Gal80 co-expression excluded the mushroom body neurons from the fru+, d5-HT1B+ population, indicated by the loss of mCD8::GFP labeling. nc82 staining is in red and marks the neuropil. Scale bar = 40 μm. (c) The fru+, d5-HT1B+ neuron in the VNC is sexually dimorphic, but does not exhibit anti-GAD staining. One fru+, d5-HT1B+ neuron is evident in the male VNC and it does not overlap with anti-GAD immunostaining. No soma was labeled in female VNC. Scale bar = 40 μm.

  13. Aggression duration measurements corresponding to Fig. 3c and Fig. 5c.
    Supplementary Fig. 7: Aggression duration measurements corresponding to Fig. 3c and Fig. 5c.

    Error bars denote s.e.m.

  14. fru+, GABA+ neurons display sexual dimorphism in the brain and VNC.
    Supplementary Fig. 8: fru+, GABA+ neurons display sexual dimorphism in the brain and VNC.

    (a) fru+, GABA+ neurons showed distinct projection patterns, as labeled by Dscam17.1::GFP, and distinct cell distribution in the VNC, as labeled by CD8::GFP between males and females. (b) The relationship between the dendrite branching pattern of fru+, Rdl+ neurons and the GABA distribution in the male fly brain and VNC. The dendritic patterning of fru+, Rdl+ neurons is shown in green using Dscam17.1::GFP and the GABA distribution is shown in red by anti-GABA staining. Scale bar = 40 μm.

  15. GABAergic inhibition underlies the female contact dependent suppression of sex-related male-male aggression.
    Supplementary Fig. 9: GABAergic inhibition underlies the female contact dependent suppression of sex-related male-male aggression.

    a) Aggression duration measurements corresponding to Fig. 6a. (b) Rdl transcripts are reduced in the hypomorphic Rdl allele RdlMB08800.Semi-quantitative RT-PCR analyses showed a reduction of Rdl mRNA level in the RdlMB08800 allele as compared to the wild type control, using primers targeting two regions of the transcript. The numbers indicate the base pair location. The levels of α;-tubulin, used as the loading control, are comparable between the two genotypes. (c, d) Aggression duration measurements corresponding to Fig. 6b. Error bars denote s.e.m. (e) A model for GABA inhibition involved in the aggression suppression. Male flies, when encountering females, activated their female pheromone sensing ppk29 neurons through direct contacts, followed by the GABAergic inhibition of central neurons in the aggression circuit, which possibly express the Rdl GABAA receptor and lead to reduced aggressive behavioral output.

Videos

  1. Supplementary Movie 1
    Video 1: Supplementary Movie 1
    Representative one minute long video clips extracted from the 60-90 min period. For all the videos, the configuration is as the following: top left-naive males only, top right-experienced males only, middle left-naive males + virgin females, middle right-experienced males + virgin females, bottom left-naive males + mated females, bottom right-experienced males + mated females. The genotypes are as follows: cs
  2. Supplementary Movie 2
    Video 2: Supplementary Movie 2
    cs; UAS>stop>TNTinactive/+; fruFLP/d5-HT1B-Gal4
  3. Supplementary Movie 3
    Video 3: Supplementary Movie 3
    cs; UAS>stop>TNTactive/+; fruFLP /d5-HT1B-Gal4
  4. Supplementary Movie 4
    Video 4: Supplementary Movie 4
    cs; UAS>stop>dTrpA1/+; fruFLP /d5-HT1B-Gal4 at 20°C
  5. Supplementary Movie 5
    Video 5: Supplementary Movie 5
    cs; UAS>stop>dTrpA1/+; fruFLP /d5-HT1B-Gal4 at 29°C

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Author information

  1. These authors contributed equally to this work.

    • Quan Yuan &
    • Yuanquan Song

Affiliations

  1. Department of Physiology and Howard Hughes Medical Institute, University of California at San Francisco, San Francisco, California, USA.

    • Quan Yuan,
    • Yuanquan Song,
    • Chung-Hui Yang,
    • Lily Yeh Jan &
    • Yuh Nung Jan
  2. Present addresses: Dendrite Morphogenesis and Plasticity Unit, National Institute of Neurological Disorders and Stroke, US National Institute of Health, Bethesda, Maryland, USA (Q.Y.), Department of Neurobiology, Duke University, Durham, North Carolina, USA (C.-H.Y.).

    • Quan Yuan,
    • Chung-Hui Yang &
    • Yuh Nung Jan

Contributions

Q.Y. and Y.S. carried out all the experiments and performed the data analysis. C.-H.Y. contributed to the behavioral assay. Q.Y., Y.S., L.Y.J. and Y.N.J. conceived the research and wrote the manuscript.

Competing financial interests

The authors declare no competing financial interests.

Corresponding author

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Supplementary information

Supplementary Figures

  1. Supplementary Figure 1: Experience with females inhibits sex-related male-male aggression. (180 KB)

    (a) Average aggression duration of cs males was quantified in 5 min intervals for 2 hours for naïve male only (blue), experienced males only (purple), naïve males + virgin females (red), experienced males + virgin females (yellow), naïve males + mated females (green), experienced males + mated females (brown). (n = 4 pairs for each condition). (b) Courtship index measured at 70–75 min for naïve male + virgin female and naive male + mated female. The latter showed significantly higher courtship index. (n = 4 pairs for each condition). (c) Average aggression frequency of cs and w1118 males was quantified for the 60-90 min. No obvious difference in the inhibition of aggression is apparent between the two genotypes. (n = 11 and 6 pairs for cs and w1118). **: P < 0.01, ***: P < 0.001 by Student's t test. Error bars denote s.e.m.

  2. Supplementary Figure 2: Experimental schemes for behavioral analysis. (221 KB)

    (a) Detailed experimental scheme for analyzing sex-related male-male aggression and its regulation by prior female experience. (b) The dis-inhibition phenotype was examined after temperature shift to induce silencing of specific group of neurons with enhancer-Gal4 driven UAS-Gal80ts and UAS-Kir2.1.

  3. Supplementary Figure 3: Conditioning parameters for the prior female experience dependent inhibition of aggression. (89 KB)

    Inhibition of aggression were displayed by males conditioned with virgin females for 24 hours, with mated females, with virgin females that express membrane bound sex peptide using fru-Gal4, which would therefore reject males, and with Drosophila pseudoobscura. However, conditioning with virgin females for 10 hours, removing females right after mating during conditioning, conditioning males in vials previously housed virgin females, conditioning males with virgin females that were separated by a nylon mesh, conditioning with Drosophila virilis, or with extra males all failed to elicit inhibition of aggression. On the other hand, removing females after mating during testing abolished aggression, revealing the presence of females during the aggression assay is crucial. (n = 11, 11, 5, 6, 6, 6, 6, 5, 5, 5, 6 and 8 pairs of flies for each condition). ***: P < 0.001 by One-way ANOVA followed by Bonferroni's multiple comparison test. Error bars denote s.e.m.

  4. Supplementary Figure 4: Previous female contact does not significantly affect the courtship index and locomotion of experienced males. Social hierarchy established between males housed in pairs has little to do with the female-contacts dependent inhibition of sex-related male aggression. (114 KB)

    (a) The courtship index and climbing index of experienced vs. naive male flies did not exhibit significant differences. (n = 5 flies for courtship index and 3 groups for climbing index). (b) Two male flies that were housed in pairs but not with each other (mixed group), so that they had social experiences but no established hierarchy with each other, exhibited as much contact-dependent inhibition of the sex-related aggression as did males that were raised together (pair). (n = 5 pairs of flies for each condition). **: P = 0.0012, by Student's t-test. . Error bars denote s.e.m.

  5. Supplementary Figure 5: Aggression duration measurements corresponding to Fig. 2a, 2b, 2d (n = 11, 6, 7 and 5 pairs of flies for each condition). (340 KB)

    ***: P < 0.001, by One-way ANOVA followed by Bonferroni's multiple comparison test.. Error bars denote s.e.m.

  6. Supplementary Figure 6: Sexual dimorphic distribution of fru+, d5-HT1B+ neurons in the brain and VNC of adult male and female flies. (1,172 KB)

    (a) fru+, d5-HT1B+ neurons in the adult male brain labeled by mCD8::GFP including γ-neurons of the mushroom bodies and a cluster with soma located below the antennal lobes and above the SOG region. Scale bar = 40 μm. The magnified view is shown in the lower left panel. Scale bar = 10 μm. nc82 co-staining in red shows the neuropil. A 3D rendering of the neuropil is shown in the lower right panel. (b) A subpopulation of fru+, d5-HT1B+ neurons independent of mushroom bodies are required for the prior female-contacts dependent inhibition of aggression. MB-Gal80 co-expression excluded the mushroom body neurons from the fru+, d5-HT1B+ population, indicated by the loss of mCD8::GFP labeling. nc82 staining is in red and marks the neuropil. Scale bar = 40 μm. (c) The fru+, d5-HT1B+ neuron in the VNC is sexually dimorphic, but does not exhibit anti-GAD staining. One fru+, d5-HT1B+ neuron is evident in the male VNC and it does not overlap with anti-GAD immunostaining. No soma was labeled in female VNC. Scale bar = 40 μm.

  7. Supplementary Figure 7: Aggression duration measurements corresponding to Fig. 3c and Fig. 5c. (110 KB)

    Error bars denote s.e.m.

  8. Supplementary Figure 8: fru+, GABA+ neurons display sexual dimorphism in the brain and VNC. (1,189 KB)

    (a) fru+, GABA+ neurons showed distinct projection patterns, as labeled by Dscam17.1::GFP, and distinct cell distribution in the VNC, as labeled by CD8::GFP between males and females. (b) The relationship between the dendrite branching pattern of fru+, Rdl+ neurons and the GABA distribution in the male fly brain and VNC. The dendritic patterning of fru+, Rdl+ neurons is shown in green using Dscam17.1::GFP and the GABA distribution is shown in red by anti-GABA staining. Scale bar = 40 μm.

  9. Supplementary Figure 9: GABAergic inhibition underlies the female contact dependent suppression of sex-related male-male aggression. (198 KB)

    a) Aggression duration measurements corresponding to Fig. 6a. (b) Rdl transcripts are reduced in the hypomorphic Rdl allele RdlMB08800.Semi-quantitative RT-PCR analyses showed a reduction of Rdl mRNA level in the RdlMB08800 allele as compared to the wild type control, using primers targeting two regions of the transcript. The numbers indicate the base pair location. The levels of α;-tubulin, used as the loading control, are comparable between the two genotypes. (c, d) Aggression duration measurements corresponding to Fig. 6b. Error bars denote s.e.m. (e) A model for GABA inhibition involved in the aggression suppression. Male flies, when encountering females, activated their female pheromone sensing ppk29 neurons through direct contacts, followed by the GABAergic inhibition of central neurons in the aggression circuit, which possibly express the Rdl GABAA receptor and lead to reduced aggressive behavioral output.

Video

  1. Video 1: Supplementary Movie 1 (2.11 MB, Download)
    Representative one minute long video clips extracted from the 60-90 min period. For all the videos, the configuration is as the following: top left-naive males only, top right-experienced males only, middle left-naive males + virgin females, middle right-experienced males + virgin females, bottom left-naive males + mated females, bottom right-experienced males + mated females. The genotypes are as follows: cs
  2. Video 2: Supplementary Movie 2 (920 KB, Download)
    cs; UAS>stop>TNTinactive/+; fruFLP/d5-HT1B-Gal4
  3. Video 3: Supplementary Movie 3 (1.01 MB, Download)
    cs; UAS>stop>TNTactive/+; fruFLP /d5-HT1B-Gal4
  4. Video 4: Supplementary Movie 4 (2.15 MB, Download)
    cs; UAS>stop>dTrpA1/+; fruFLP /d5-HT1B-Gal4 at 20°C
  5. Video 5: Supplementary Movie 5 (2.19 MB, Download)
    cs; UAS>stop>dTrpA1/+; fruFLP /d5-HT1B-Gal4 at 29°C

PDF files

  1. Supplementary Text and Figures (5,853 KB)

    Supplementary Figures 1–9, Supplementary Table 1 and Supplementary Movies 1–5

Image files

  1. Supplementary Table 1 (337 KB)

    Summary of the enhancer-Gal4 lines tested with tub-Gal80ts; UAS-Kir2.1.

Additional data