Figures and data in Sex-determining genes distinctly regulate courtship capability and target preference via sexually dimorphic neurons

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6 figures, 8 videos, 1 table and 3 additional files

Figures

Figure 1 with 3 supplements

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Sex of the target fly influences behaviors triggered by the optogenetic activation of social behavior-promoting neurons.

(A) Expression of CsChrimson:tdTomato under the control of *dsx^GAL4* and *fru^FLP* (red in A₁, black in **A_2,3**) in a male brain is visualized together with a neuropil marker BRP (blue in A₁) by immunohistochemistry. Labeled cell body clusters are enlarged in A₃. Scale bar: 100 μm (A₁), 10 μm (A₃). (B) Mean number of cell bodies per hemibrain visualized by anti-DsRed antibody in male (left) and female (right) brains. (C) Schematics of the design of behavioral assays. (D) Schematics of the optogenetic stimulation paradigm. Time windows 1–4 represent periods in which behavioral parameters are pooled and calculated in subsequent panels. (**E, G**) Rasters of behaviors (indicated in left) performed by male tester flies that express CsChrimson:tdTomato under the control of *dsx^GAL4* and *fru^FLP*. Pink bar: LED-on periods, horizontal bar: 10 min (also see D), vertical bar: 10 flies. LED stimulation condition is indicated at the bottom. (**F, H**): Boxplots of time orienting (**F₁, H₁**), lunges (**F₂, H₂**), and wing extension (**F₃, H₃**) by the tester flies during the time windows 1–4 (see D). Testers’ genotypes and pair numbers are indicated below the plots. Gray lines represent single testers. In gray: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Wilcoxon signed rank test). In black: **p<0.01, n.s., p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test). Target flies are either wild-type group-housed males (**E, F**) or mated females (**G, H**). (I) Comparison of time orienting (I₁), lunges (I₂), and wing extension (I₃) performed by tester flies that express CsChrimson:tdTomato under the control of *dsx^GAL4* and *fru^FLP* in males (data replotted from F, H) toward male or female target flies (indicated above). Number of pairs tested and time windows compared are indicated below the panels. **p<0.01 (Mann-Whitney U-test). For detailed methods to quantify behaviors, see Materials and methods.

Figure 1—figure supplement 1

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Additional characterization of the dsx^GAL4 ∩ fru^FLP neurons.

(**A, B**) Expression of CsChrimson:tdTomato under the control of *dsx^GAL4* and *fru^FLP* (red in A₁, B₁, black in A₂, **B_2,3**) in a male ventral nerve cord (A) and a female brain (B) is visualized together with a neuropil marker BRP (blue in A₁, B₁) by immunohistochemistry. Labeled cell body clusters in B₂ are enlarged in B₃). Scale bar: 100 μm (A₁, B₁), 10 μm (B₃).

Figure 1—figure supplement 2

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Performance of automated behavior classifiers.

(A) Precision and recall values of the three behavioral classifiers used in this study. See Materials and methods for the definition of precision and recall. (B) Boxplots of lunges (B₁), wing extensions (B₂), and headbutts (B₃), performed by pairs of wild-type male flies (singly reared for 6 days) (left), pairs of a wild-type male and a wild-type mated female (reared in same-sex groups for 6 days) (middle), and pairs of wild-type virgin female flies (singly reared for 6 days) (right) during 30 min assays. For male pairs and virgin female pairs, data from marked flies (whose wing tips were clipped) are shown on the right side. Pair numbers are indicated below the panel.

Figure 1—figure supplement 3

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Behaviors induced by optogenetic stimulation of dsx^GAL4 ∩ fru^FLP neurons across various LED frequencies.

(**A, B**) Boxplots of time orienting (**A₁, B₁**), lunges (**A₂, B₂**) and wing extension (**A₃, B₃**) by the tester flies during the time windows 1–4 (see Figure 1D). Tester’s genotypes and pair numbers are indicated below the plots. Gray lines represent single testers. LED frequencies were varied as indicated below the plot. Target flies are wild-type group-housed males (A) or mated females (B). In gray: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Wilcoxon signed rank test). In black: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test).

Figure 2 with 3 supplements

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NP2631 ∩ dsx^FLP neurons that express both *dsx* and *fru* can promote courtship and aggressive behaviors.

(**A–C**): Boxplots of time orienting (**A₁–C₁**), lunges (**A₂–C₂**), and wing extension (**A₃–C₃**) by the tester flies during the time windows 1–4 (see Figure 1D). Testers’ genotypes and pair numbers are indicated below the plots. Gray lines represent single testers. Target flies are either group-housed wild-type males (**A, C**) or mated females (B). Gray lines represent single testers. In gray: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Wilcoxon signed rank test). In black: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test). (D). A schematic summary. A subset of NP2631 ∩ dsx^FLP neurons that express *fru* (labeled by the *fru^P1.LexA* allele) (orange) can promote both lunges and wing extensions.

Figure 2—figure supplement 1

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Detailed description of behaviors induced by the optogenetic activation of NP2631 ∩ dsx^FLP neurons.

(**A, B**) Rasters of time orienting (**A₁, B₁**), lunges (**A₂, B₂**), and wing extensions (**A₃, B₃**) performed by male tester flies that express CsChrimson:tdTomato under the control of *NP2631* and *dsx^FLP* (the same datasets as used in Figure 2A, (B). Pink bar: LED-on periods, horizontal bar: 10 min, vertical bar: 10 flies. LED stimulation condition is indicated at the bottom. (C): Boxplots of time orienting (C₁), lunges (C₂), and wing extension (C₃) by the tester flies during the time windows 2 (left) and 4 (right) (see Figure 1D). Tester’s genotype (indicated below) is the same as testers in A) and B). LED frequencies were varied as indicated below the plot. Target flies are wild-type group-housed males. (D) Boxplots of velocity (D₁), lunges (D₂), and wing extensions (D₃) performed by the tester flies that express CsChrimson:tdTomato under the control of *NP2631* and *dsx^FLP* in the absence of a target fly. Testers’ genotypes and pair numbers are indicated below the plots. .

Figure 2—figure supplement 2

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A detailed characterization of NP2631 ∩ dsx^FLP neurons.

(A): Expression of CsChrimson:tdTomato in a representative male central nervous system (brain and ventral nerve cord (VNC)) under the control of *NP2631* and *dsx^FLP* is visualized by immunohistochemistry (scale bar: 100 µm). The P1/pC1 clusters at the posterior part of the brains are indicated in orange circles. The majority of brains had no other labeled neurons (A₁), but 35% (see D₁) of the examined brains (A₂) also had a single neuron with a large cell body (brown arrowhead) that extend contralateral and descending projections (open brown arrowheads). (B) Representative image of NP2631 ∩ dsx^FLP neurons (red in **B₁–B₃**), black in B₄) visualized by immunohistochemistry against tdTomato along with immunoreactivity against DsxM (yellow in B_{1, 2}, black in B₅) and FruM (cyan in B_{1, 3}, black in B₆) (scale bar: 10 µm). Two CsChrimson:tdTomato-expressing neurons also express both DsxM and FruM (green circles in B_{5, 6}) whereas four other neurons express DsxM only (cyan circles in B_{5, 6}). (C) Mean number of P1/pC1 NP2631 ∩ dsx^FLP neuronal cell bodies per hemibrain, categorized by immunoreactivity against DsxM and FruM. Gray lines represent single hemibrain. Error bars: S.D. (D) Pie charts that shows ratios of flies with a NP2631 ∩ dsx^FLP descending neuron (brown in D₁), flies that increased lunges toward male targets during LED stimulation (window two in Figure 1D) relative to during pre-stimulation (window one in Figure 1D) (blue in D₂) left), or during inter-stimulus intervals (window three in Figure 1D) (blue in D₂) right), flies that increased wing extensions toward female targets in window two relative to in window 1 (pink in D₃) left), or in window 3 (pink in D₃) right), among male flies that express CsChrimson:tdTomato under the control of *NP2631* and *dsx^FLP*. Behavioral data are from the same data set used in Figure 3—figure supplement 1I2 (D₂) and I₃ (D₃), **p<0.01, Fisher’s exact test for the frequency of a descending neuron. (**E–G**) Expression of CsChrimson:tdTomato (red in **E₁–G₁**), black in **E₂–G₂**) in a representative male brain in genotypes shown in H) is visualized together with the neuropil marker BRP (blue in **E₁–G₁**) by immunohistochemistry (scale bar: 100 µm). (H): Mean number of cell bodies per hemibrain visualized by anti-DsRed antibody in each genotype shown in left. Brains from the top genotype were also co-immunostained by an anti-FruM antibody, and the breakdown of FruM-expressing cells and FruM-negative cells is also shown (gray lines represent single hemibrains). Error bars: S.D. Number of hemibrains examined are indicated to the left. Lowercase letters denote significance group (p<0.05, one-way ANOVA with post-hoc Tukey’s honestly significant difference test). n.s. p>0.05, Student’s t-test.

Figure 2—figure supplement 3

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Behaviors of target flies remain largely unaffected by the optogenetic stimulation of NP2631 ∩ dsx^FLP neurons in tester flies.

(**A, B**) Boxplots of time orienting (**A₁, B₁**), lunges (A₂), headbutts (B₂), and wing extension (**A₃, B₃**) by the target male (A) or female (B) flies during the time windows 4 (see Figure 1D) of the experiments shown in Figure 2A (A) and 2B (B). Testers’ genotypes and pair numbers are indicated below the plots. **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test).

Figure 3 with 1 supplement

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*dsx* specifies the sexual dimorphism of NP2631 ∩ dsx^FLP neurons.

(A) Schematics of the sex-determination pathway in *Drosophila*. (B) Schematic of the four sex genotypes defined by *dsx* and *fru* splicing, and how NP2631 ∩ dsx^FLP neurons are specified in each genotype (see following panels for details). (**C–F**) Expression of CsChrimson:tdTomato under the control of *NP2631* and *dsx^FLP* (red in **C₁–F₁**, black in C_2,3-F_2,3) in brains of a male (C), fruM female (D), fruF male (E), and female (F) is visualized together with a neuropil marker BRP (blue) by immunohistochemistry. Circle: soma (right cluster is enlarged in **C₃–F₃**). Scale bar: 100 μm (**C₁–F₁**), 10 μm (**C₃–F₃**). (G) Mean number of cell bodies per hemibrain visualized by anti-DsRed antibody in each genotype represented in **C–F**) and Figure 3—figure supplement 1A–D. (**H–K**) Z-projection of the registered and averaged images of CsChrimson:tdTomato expression under the control of *NP2631* and *dsx^FLP* (black in **H₁–K₁**) in male (H), fruM female (I), fruF male (J), and female (K). A part of the standard *Drosophila* brain is shown in gray in **H₂–K₂**). Number of used hemibrains are indicated in **H₂–K₂**). Lateral junction (orange), lateral vertical projection (green), and superior medial projection (magenta) are segmented and overlaid in H₂-K₂. L-M: Boxplot of volumes of lateral junction (L), lateral vertical projection (M), and superior medial projections (N). Genotypes and number of hemibrains are indicated below the plot. **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Wilcoxon signed rank test).

Figure 3—figure supplement 1

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Heterozygosity of the *fru^4-40* allele leaves the neuroanatomy and function of NP2631 ∩ dsx^FLP neurons largely unaltered.

(**A–D**) Expression of CsChrimson:tdTomato under the control of *NP2631* and *dsx^FLP* (red in **A₁–D₁**), black in A_2,3-D_2,3) in representative brains of a *fru* wild-type male (A), *fru* wild-type female (B), *fru^4-40* heterozygous male (C), and *fru^4-40* heterozygous female (D), is visualized together with a neuropil marker BRP (blue in **A₁–D₁**) by immunohistochemistry. Scale bar: 100 μm. Circle: soma. (**E–H**) Z-projection of the registered and averaged images of CsChrimson:tdTomato expression under the control of *NP2631* and *dsx^FLP* in *fru* wild-type male (E), *fru^M*/*fru^4-40* male (F), duplication from Figure 3H), *fru* wild-type female (G), and *fru^F*/*fru^4-40* female (H), duplication from Figure 3K). Number of used hemibrains are indicated. (I) Boxplots of time orienting (I₁), lunges (I₂), and wing extensions (I₃) performed by male tester flies that express CsChrimson:tdTomato under the control of *NP2631* and *dsx^FLP*. Only values during the time window four are shown. Testers’ *fru* locus genotypes and pair numbers are indicated below the plots. Target flies are either group-housed wild-type males or mated females, as indicated. Plots with gray shades are replots of the data sets shown in Figure 2A and B. **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test).

Figure 4 with 1 supplement

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NP2631 ∩ dsx^FLP neurons in fruF males promote courtship, but not aggressive, behavior.

(**A, B**) Boxplots of time orienting (**A₁, B₁**), lunges (**A₂, B₂**), and wing extension (**A₃, B₃**) by the tester flies during the time windows 1–4 (see Figure 1D). Testers’ genotypes and pair numbers are indicated below the plots. Gray lines represent single testers. Target flies are either group-housed wild-type males (A) or mated females (B). In gray: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Wilcoxon signed rank test). In black: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test). (C) Comparison of wing extension performed by flies that express CsChrimson:tdTomato under the control of *NP2631* and *dsx^FLP* in males (data from Figure 3—figure supplement 1I) or in fruF males (data from A, (B), during the time window 4. Sex of tester and target flies is indicated above the panels. Number of pairs tested is indicated below the panels. **p<0.01, n.s. p>0.05 (Mann-Whitney U-test). (D) Models of the function of NP2631 ∩ dsx^FLP neurons in the context of *dsx* and *fru*.

Figure 4—figure supplement 1

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Rasters of behaviors induced by the optogenetic activation of NP2631 ∩ dsx^FLP neurons in fruF males.

(**A, B**) Rasters of time orienting (**A₁, B₁**), lunges (**A₂, B₂**), and wing extensions (**A₃, B₃**) performed by fruF male tester flies that express CsChrimson:tdTomato under the control of *NP2631* and *dsx^FLP* (the same datasets as used in Figure 4A, (B). Pink bar: LED-on periods, horizontal bar: 10 min, vertical bar: 10 flies. LED stimulation condition is indicated at the bottom.

Figure 5 with 3 supplements

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P1^a neurons are distinct from NP2631 ∩ dsx^FLP neurons.

(**A, C**) Expression of CsChrimson:tdTomato under the control of *NP2631* and *dsx^FLP* (yellow), GCaMP6f under the control of *R15A01-LexA* (green in A) or *R71G01-LexA* (green in C), and a neuropil marker BRP (blue) in a representative male brain are visualized by immunohistochemistry. Scale bar: 100 μm. An inset represents a magnified view of the posterior cell body cluster in the white rectangle (scale bar: 10 μm). An open arrowhead in C) indicates a neural tract from LexA-expressing neurons that appear distinct from the tract from neurons labeled by *NP2631* and *dsx^FLP* (white arrowhead). (**B, D**) Mean number of cell bodies per hemibrain with immunohistochemical signal by anti-DsRed antibody (orange), anti-GFP antibody (green), and both antibodies (purple) in brains of the genotype represented in A (B) or in C (D). In D, all LexA-expressing neurons located near *NP2631* and *dsx^FLP* neurons are included, although some may belong to different neuronal clusters. Error bars: S.D. Gray lines represent single hemibrains. (**E–G**) Z-projection of the registered and averaged images of CsChrimson:tdTomato expression under the control of *P1^a-GAL4* (cyan in **E₁, G**, black in E₂), and *NP2631* and *dsx^FLP* (orange in **F₁, G**, black in F₂; duplication from Figure 3H) neurons. A part of the standard *Drosophila* brain is shown in gray. Number of hemibrains used are indicated in E₂ and F₂.

Figure 5—figure supplement 1

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Detailed characterization of P1^a neurons.

(**A, B**) Boxplots of time orienting (**A₁, B₁**), lunges (**A₂, B₂**), and wing extension (**A₃, B₃**) by the tester flies during the time windows 1–4 (see Figure 1D). Testers’ genotypes and pair numbers are indicated below the plots. Gray lines represent single testers. Target flies are either group-housed wild-type males (A) or mated females (B). Gray lines represent single testers. In gray: **p<0.01, *p<0.05, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Wilcoxon signed rank test). In black: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test). (C) Representative image of P1^a neurons (red in **C₁–C₃**, black in C₄) visualized by immunohistochemistry against tdTomato along with immunoreactivity against DsxM (yellow in C_{1, 2}, black in C₅) and FruM (cyan in C_{1, 3}, black in C₆) (scale bar: 10 µm). Eight CsChrimson:tdTomato-expressing neurons also express both DsxM and FruM (green circles in C_{5, 6}) whereas two other neurons express DsxM only (cyan circles in C_{5, 6}). (D) Mean number of P1^a neuronal cell bodies per hemibrain, categorized by immunoreactivity against DsxM and FruM. Gray lines represent single hemibrains. Error bars: S.D.

Figure 5—figure supplement 2

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Detailed characterization of P1^a ∩ fru^FLP neurons.

(A) Mean number of cell bodies per hemibrain visualized by anti-DsRed antibody in each genotype shown in left. Brains from the top genotype were also co-immunostained by an anti-FruM antibody, and the breakdown of FruM-expressing cells and FruM-negative cells is also shown (gray lines represent single hemibrains). Error bars: S.D. Number of hemibrains examined are indicated to the left. Lowercase letters denote significance group (p<0.05, one-way ANOVA with post-hoc Tukey’s honestly significant difference test). n.s. p>0.05, Student’s t-test. (B) Boxplots of time orienting (B₁), lunges (B₂), and wing extension (B₃) by the tester flies during the time windows 1–4 (see Figure 1D). Testers’ genotypes and pair numbers are indicated below the plots. Gray lines represent single testers. Target flies are group-housed wild-type males. Gray lines represent single testers. In gray: **p<0.01, *p<0.05, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Wilcoxon signed rank test). In black: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test). (**C–E**) Z-projection of the registered and averaged images of CsChrimson:tdTomato expression under the control of *P1^a-GAL4* and *fru^FLP* (cyan in C, E) and *NP2631* and *dsx^FLP* (orange in **D, E**; duplication from Figure 3—figure supplement 1E) neurons. A part of the standard *Drosophila* brain is shown in gray. Number of used hemibrains are indicated in C₂ and D₂. Note that the genotype of animals in B (cyan) and C is identical to the 3^rd genotype from the top in A.

Figure 5—figure supplement 3

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P1^a and NP2631 ∩ fru^FLP neurons do not overlap.

(**A, C**) Expression of CsChrimson:tdTomato under the control of *NP2631* and *fru^FLP* (cyan), GCaMP6f under the control of *R15A01-LexA* (green in A) or *R71G01-LexA* (green in C), and a neuropil marker BRP (blue) in a representative male brain are visualized by immunohistochemistry. Scale bar: 100 μm. An inset represents magnified view of the posterior cell body cluster in the white rectangle (scale bar: 10 μm). (**B, D**) Mean number of cell bodies per hemibrain with immunohistochemical signal by anti-DsRed antibody (cyan), anti-GFP antibody (green), and both antibodies (purple) in brains of the genotype represented in A (B) or in C (D). Error bars: S.D. Gray lines represent single hemibrains.

Figure 6 with 2 supplements

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Both *dsx* and *fru* specify the sexual dimorphism of P1^a neurons.

(**A–D**) Expression of CsChrimson:tdTomato under the control of *P1^a-GAL4* (red in **A₁–D₁**, black in A_2,3-D_2,3) in brains of a male (A), fruM female (B), fruF male (C), and female (D) is visualized together with a neuropil marker BRP (blue) by immunohistochemistry. Circle: soma (right cluster is enlarged in A₃ and C₃), red asterisk: sex-invariant background labeling (see Materials and methods for details). Scale bar: 100 μm (**A₁–D₁**), 10 μm (**A₃, C₃**). (E) Mean number of cell bodies per hemibrain visualized by anti-DsRed antibody in each genotype represented in **A–D**) and Figure 6—figure supplement 1A–D. (**F, G**) Z-projection of the registered and averaged images of CsChrimson:tdTomato expression under the control of *P1^a-GAL4* in male (F) and fruF male (G). A part of the standard *Drosophila* brain is shown in gray in **F₂, G₂**. Number of hemibrains used are indicated in F₂) and G₂). Lateral junction (cyan) and dorsal posterior projection (yellow) are segmented and overlaid in F₂) and G₂. H, I. Boxplot of volumes of lateral junction (H) and dorsal posterior projection (I). Genotypes and number of hemibrains are indicated below the plot. **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test).

Figure 6—figure supplement 1

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Heterozygosity of the *fru^4-40* allele leaves the neuroanatomy and function of P1^a neurons largely unaltered.

(**A–D**) Expression of CsChrimson:tdTomato under the control of *P1^a* (red in **A₁–D₁**), black in A_2,3-D_2,3) in representative brains of a *fru* wild-type male (A), *fru* wild-type female (B), *fru^4-40* heterozygous male (C), and *fru^4-40* heterozygous female (D), is visualized together with a neuropil marker BRP (blue in **A₁–D₁**) by immunohistochemistry. Scale bar: 100 μm. Circle: soma, red asterisk: sex-invariant background labeling.( **E, F**) Z-projection of the registered and averaged images of CsChrimson:tdTomato expression under the control of *P1^a* in *fru* wild-type male (E) duplication from Figure 5E) and *fru^M*/*fru^4-40* male (F); duplication from Figure 6F). Number of used hemibrains are indicated. (G) Boxplots of time orienting (G₁), lunges (G₂), and wing extensions (G₃) performed by male tester flies that express CsChrimson:tdTomato under the control of *P1^a*. Only values during the time window four are shown (see Figure 1D). Testers’ *fru* locus genotypes and pair numbers are indicated below the plots. Target flies are either group-housed wild-type males or mated females, as indicated. Plots with gray shades are replots of the data sets shown in Figure 5—figure supplement 1A,B. **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test).

Figure 6—figure supplement 2

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P1^a neurons in fruF males are defective in promoting both courtship and aggressive behavior.

(**A, B**) Boxplots of time orienting (**A₁, B₁**), lunges (**A₂, B₂**), and wing extension (**A₃, B₃**) by the tester flies during the time windows 1–4 (see Figure 1D). Their genotypes and numbers are indicated below the plots. Gray lines represent single testers. Target flies are either group-housed wild-type males (A) or mated females (B). In gray: **p<0.01, *p<0.05, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Wilcoxon signed rank test). In black: **p<0.01, n.s. p>0.05 (Kruskal-Wallis one-way ANOVA and post-hoc Mann-Whitney U-test). (C) Comparison of wing extension performed by flies that express CsChrimson:tdTomato under the control of *P1^a* in males (data from Figure 6—figure supplement 1G) or in fruF males (data from A, (B), during the time window 4. Sex of tester and target flies is indicated above the panels. Number of pairs tested is indicated below the panels. **p<0.01, n.s. p>0.05 (Mann-Whitney U-test).

Videos

Video 1

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posterframe for video — Representative behavior of a male tester fly that expresses.

CsChrimson:tdTomato under the control of *dsx^GAL4* and *fru^FLP* toward a wild-type male (Part 1) or a wild-type female (Part 2) target fly, at the onset and offset of LED stimulation.

Video 2

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Video 3

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Video 4

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Video 5

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Video 6

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Video 8

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Tables

Table 1

Complete genotypes of Drosophila strains used in this study.

Figure	Panel	Abbreviated genotype	COMPLETE GENOTYPE (‘Y’ represents the Y chromosome)
Figure 1	A, B	dsx^GAL4 ∩ fru^FLP ♂	w/Y; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022/+; dsx^GAL4/fru^FLP
Figure 1—figure supplement 1	A
Video 1
Figure 1	B	dsx^GAL4 ∩ fru^FLP ♀	w/w; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022/+; dsx^GAL4/fru^FLP
Figure 1—figure supplement 1	B	dsx^GAL4 ∩ fru^FLP ♀
Figure 1	E-I	dsx^GAL4, UAS > stop > CsChrimson, fru^FLP	w/Y; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022/+; dsx^GAL4/fru^FLP
Figure 1—figure supplement 1	C, D	dsx^GAL4, UAS > stop > CsChrimson, fru^FLP
Figure 1	E-H	dsx^GAL4, UAS > stop > CsChrimson	w/Y; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022/+; dsx^GAL4/+
Figure 1—figure supplement 1	C, D	dsx^GAL4, UAS > stop > CsChrimson
Figure 1	E-H	UAS > stop > CsChrimson, fru^FLP	w/Y; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022/+; fru^FLP/+
Figure 1—figure supplement 1	C, D	UAS > stop > CsChrimson, fru^FLP
Figure 1—figure supplement 2	B	Canton-S ♂	+/Y; +/+; +/+ (Canton-S)
Figure 1—figure supplement 2	B	Canton-S ♀	+/+; +/+; +/+ (Canton-S)
Figure 2	A, B	NP2631, UAS > stop > CsChrimson, dsx^FLP	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/+
Figure 2—figure supplement 1	A-C
Figure 2—figure supplement 3	A, B
Video 2
Figure 2	A, B	NP2631, UAS > stop > CsChrimson	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; fru^4-40/+
Figure 2—figure supplement 3	A, B	NP2631, UAS > stop > CsChrimson
Figure 2	A, B	UAS > stop > CsChrimson, dsx^FLP	w/Y; +/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/+
Figure 2—figure supplement 3	A, B	UAS > stop > CsChrimson, dsx^FLP
Figure 2	C	NP2631, UAS > stop > CsChrimson, dsx^FLP, fru^{P1. LexA}, LexAop2-GAL80	w/Y; NP2631/8XLexAop2-GAL80 in attP40; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, dsx^FLP/fru^{P1. LexA}
Figure 2	C	NP2631, UAS > stop > CsChrimson, dsx^FLP, LexAop2-GAL80	w/Y; NP2631/8XLexAop2-GAL80 in attP40; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, dsx^FLP/+
Figure 2	C	NP2631, UAS > stop > CsChrimson, dsx^FLP, fru^{P1. LexA}	w/Y; NP2631/+; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, dsx^FLP/fru^{P1. LexA}
Figure 2—figure supplement 1	A-C	NP2631, UAS > stop > CsChrimson, dsx^FLP	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/+
Figure 2—figure supplement 2	A-D	NP2631 ∩ dsx^FLP	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP/+
Figure 2—figure supplement 2	E, H	NP2631, UAS > stop > CsChrimson (VK00005), dsx^FLP, fru^{P1. LexA}, LexAop2-GAL80	w/Y; NP2631/8XLexAop2-GAL80 in attP40; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, dsx^FLP/fru^{P1. LexA}
Figure 2—figure supplement 2	F, H	NP2631, UAS > stop > CsChrimson (VK00005), dsx^FLP, LexAop2-GAL80	w/Y; NP2631/8XLexAop2-GAL80 in attP40; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, dsx^FLP/+
Figure 2—figure supplement 2	G, H	NP2631, UAS > stop > CsChrimson (VK00005), dsx^FLP, fru^{P1. LexA}	w/Y; NP2631/+; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, dsx^FLP/fru^{P1. LexA}
Figure 3	C, G, H, L-N	NP2631 ∩ dsx^FLP, XY, fru locus: fru^M/fru^4-40	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/fru^M
Figure 3—figure supplement 1	F
Video 3, 4
Figure 3	D, G, I, L-N	NP2631 ∩ dsx^FLP, XX, fru locus: fru^M/fru^4-40	w/w; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/fru^M
Video 5		NP2631 ∩ dsx^FLP, XX, fru locus: fru^M/fru^4-40
Figure 3	E, G, J, L-N	NP2631 ∩ dsx^FLP, XY, fru locus: fru^F/fru^4-40	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/fru^F
Video 4		NP2631 ∩ dsx^FLP, XY, fru locus: fru^F/fru^4-40
Figure 3	F, G, K-N	NP2631 ∩ dsx^FLP, XX, fru locus: fru^F/fru^4-40	w/w; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/fru^F
Figure 3—figure supplement 1	H
Video 3, 5
Figure 3	G	NP2631 ∩ dsx^FLP, XY, fru locus: +/+	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP/+
Figure 3—figure supplement 1	A, E	NP2631 ∩ dsx^FLP, XY, fru locus: +/+
Figure 3	G	NP2631 ∩ dsx^FLP, XX, fru locus: +/+	w/w; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP/+
Figure 3—figure supplement 1	B, G	NP2631 ∩ dsx^FLP, XX, fru locus: +/+
Figure 3	G	NP2631 ∩ dsx^FLP, XY, fru locus: +/fru^4-40	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/+
Figure 3—figure supplement 1	C	NP2631 ∩ dsx^FLP, XY, fru locus: +/fru^4-40
Figure 3	G	NP2631 ∩ dsx^FLP, XX, fru locus: +/fru^4-40	w/w; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/+
Figure 3—figure supplement 1	D	NP2631 ∩ dsx^FLP, XX, fru locus: +/fru^4-40
Figure 3—figure supplement 1	I	NP2631, UAS > stop > CsChrimson, dsx^FLP, fru locus: +/+	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP/+
Figure 3—figure supplement 1	I	NP2631, UAS > stop > CsChrimson, dsx^FLP, fru locus: +/fru^4-40	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/+
Figure 3—figure supplement 1	I	NP2631, UAS > stop > CsChrimson, dsx^FLP, fru locus: fru^M/fru^4-40	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/fru^M
Figure 4	A-C	fruF ♂, NP2631, UAS > stop > CsChrimson, dsx^FLP	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/fru^F
Figure 4—figure supplement 1	A, B
Video 6
Figure 4	A, B	fruF ♂, NP2631, UAS > stop > CsChrimson	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; fru^4-40/fru^F
Figure 4	A, B	fruF ♂, UAS > stop > CsChrimson, dsx^FLP	w/Y; +/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/fru^F
Figure 4	C	♂, NP2631, UAS > stop > CsChrimson,dsx^FLP	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/fru^M
Figure 5	A, B	NP2631 ∩ dsx^FLP, R15A01-LexA	w/Y; NP2631/13XLexAop2-IVS-GCaMP6f-p10 in su(Hw)attP5; R15A01-LexA in attP2/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, dsx^FLP
Figure 5	C, D	NP2631 ∩ dsx^FLP, R71G01-LexA	w/Y; NP2631/13XLexAop2-IVS-GCaMP6f-p10 in su(Hw)attP5; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, dsx^FLP/R71G01-LexA in attP2
Figure 5	E, G	P1^a-GAL4	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2/+
Video 7		P1^a-GAL4
Figure 5	F, G	NP2631 ∩ dsx^FLP	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP, fru^4-40/fru^M
Video 7		NP2631 ∩ dsx^FLP
Figure 5—figure supplement 1	A	P1^a-GAL4, UAS-CsChrimson	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2/+
Figure 5—figure supplement 1	A	P1^a-GAL4	w/Y; R15A01-p65AD:Zp in attP40/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2/+
Figure 5—figure supplement 1	A	UAS-CsChrimson	w/Y; +/20XUAS-IVS-CsChrimson:tdTomato in VK00022; +/+
Figure 5—figure supplement 1	B	P1^a-GAL4, UAS-CsChrimson	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/fru^M
Figure 5—figure supplement 1	B	P1^a-GAL4	w/Y; R15A01-p65AD:Zp in attP40/10XUAS-IVS-GFP-p10 in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/fru^M
Figure 5—figure supplement 1	B	UAS-CsChrimson	w/Y; +/20XUAS-IVS-CsChrimson:tdTomato in VK00022; fru^4-40/fru^M
Figure 5—figure supplement 1	C, D	P1^a-GAL4	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2/+
Figure 5—figure supplement 2	A	P1^a-GAL4, UAS-CsChrimson (VK00022)	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp;GAL4DBD in attP2/+
Figure 5—figure supplement 2	A	P1^a-GAL4, UAS > stop > CsChrimson (VK00022), fru^FLP	w/Y; R15A01-p65AD:Zp in attP40/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; R71G01-Zp;GAL4DBD in attP2/fru^FLP
Figure 5—figure supplement 2	A	P1^a-GAL4, UAS > stop > CsChrimson (VK00005), fru^FLP	w/Y; R15A01-p65AD:Zp in attP40/+; 71G01-Zp;GAL4DBD in attP2/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, fru^FLP
Figure 5—figure supplement 2	A	P1^a-GAL4, UAS > stop > CsChrimson (VK00005), dsx^FLP	w/Y; R15A01-p65AD:Zp in attP40/+; 71G01-Zp;GAL4DBD in attP2/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, dsx^FLP
Figure 5—figure supplement 2	C, E	P1^a-GAL4 ∩ fru^FLP	w/Y; R15A01-p65AD:Zp in attP40/+; 71G01-Zp;GAL4DBD in attP2/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, fru^FLP
Figure 5—figure supplement 2	D, E	NP2631 ∩ dsx^FLP	w/Y; NP2631/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00022; dsx^FLP/+
Figure 5—figure supplement 2	B	P1^a-GAL4,UAS > stop > CsChrimson, fru^FLP	w/Y; R15A01-p65AD:Zp in attP40/+;R71G01-Zp:GAL4DBD in attP2/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, fru^FLP
Figure 5—figure supplement 2	B	P1^a-GAL4, UAS > stop > CsChrimson	w/Y; R15A01-p65AD:Zp in attP40/+; R71G01-Zp;GaL4DBD in attP2/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005
Figure 5—figure supplement 2	B	UAS > stop > CsChrimson,fru^FLP	w/Y; R15A01-p65AD:Zp in attP40/+; +/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, fru^FLP
Figure 5—figure supplement 3	A, B	NP2631 ∩ fru^FLP, R15A01-LexA	w/Y; NP2631/13XLexAop2-IVS-GCaMP6f-p10 in su(Hw)attP5; R15A01-LexA in attP2/20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, fru^FLP
Figure 5—figure supplement 3	C, D	NP2631 ∩ fru^FLP, R71G01-LexA	w/Y; NP2631/13XLexAop2-IVS-GCaMP6f-p10 in su(Hw)attP5; 20XUAS > myr:TopHAT2 > CsChrimson:tdTomato in VK00005, fru^FLP/R71G01-LexA in attP2
Figure 6	A, E, F, H, I	P1^a-GAL4, XY, fru locus: fru^M/fru^4-40	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/fru^M
Figure 6—figure supplement 1	F	P1^a-GAL4, XY, fru locus: fru^M/fru^4-40
Figure 6	B, E	P1^a-GAL4, XX, fru locus:fru^M/fru^4-40	w/w; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/fru^M
Figure 6	C, E, G-I	P1^a-GAL4, XY, fru locus: fru^F/fru^4-40	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/fru^F
Figure 6	D, E	P1^a-GAL4, XX, fru locus:fru^F/fru^4-40	w/w; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/fru^F
Figure 6	E, H, I	P1^a-GAL4, XY, fru locus: +/+	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2/+
Figure 6—figure supplement 1	A, E	P1^a-GAL4, XY, fru locus: +/+
Figure 6	E	P1^a-GAL4, XX, fru locus: +/+	w/w; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2/+
Figure 6—figure supplement 1	B	P1^a-GAL4, XX, fru locus: +/+
Figure 6	E	P1^a-GAL4, XY, fru locus: +/fru^4-40	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/+
Figure 6—figure supplement 1	C	P1^a-GAL4, XY, fru locus: +/fru^4-40
Figure 6	E	P1^a-GAL4, XX, fru locus: +/fru^4-40	w/w; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/+
Figure 6—figure supplement 1	D	P1^a-GAL4, XX, fru locus: +/fru^4-40
Figure 6—figure supplement 1	G	P1^a-GAL4, UAS-CsChrimson, fru locus: +/+	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, +/+
Figure 6—figure supplement 1	G	P1^a-GAL4, UAS-CsChrimson, fru locus: +/fru^4-40	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/+
Figure 6—figure supplement 1	G	P1^a-GAL4, UAS-CsChrimson, fru locus: fru^M/fru^4-40	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/fru^F
Figure 6—figure supplement 2	A-C	fruF ♂, P1^a-GAL4, UAS-CsChrimson	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/fru^F
Figure 6—figure supplement 2	A	fruF ♂, P1^a-GAL4	w/Y; R15A01-p65AD:Zp in attP40/10XUAS-IVS-GFP-p10 in VK00022; R71G01-Zp:GAL4DBD in attP2, fru^4-40/fru^F
Figure 6—figure supplement 2	A	fruF ♂, UAS-CsChrimson	w/Y; R15A01-p65AD:Zp in attP40/20XUAS-IVS-CsChrimson:tdTomato in VK00022; fru^4-40/fru^F
Figure 6—figure supplement 2	C	♂, P1^a-GAL4, UAS-CsChrimson

Additional files

Source data 1 This file contains data points represented in all figure panels, as well as p-values for all statistical tests shown in the figures.: https://cdn.elifesciences.org/articles/52701/elife-52701-data1-v1.xlsx
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Supplementary file 1 Key Resources Table.: https://cdn.elifesciences.org/articles/52701/elife-52701-supp1-v1.docx
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Kenichi Ishii
Margot Wohl
Andre DeSouza
Kenta Asahina

(2020)

Sex-determining genes distinctly regulate courtship capability and target preference via sexually dimorphic neurons

eLife 9:e52701.

https://doi.org/10.7554/eLife.52701

Figures

Sex of the target fly influences behaviors triggered by the optogenetic activation of social behavior-promoting neurons.

Additional characterization of the dsx^GAL4 ∩ fru^FLP neurons.

Performance of automated behavior classifiers.

Behaviors induced by optogenetic stimulation of dsx^GAL4 ∩ fru^FLP neurons across various LED frequencies.

NP2631 ∩ dsx^FLP neurons that express both dsx and fru can promote courtship and aggressive behaviors.

Detailed description of behaviors induced by the optogenetic activation of NP2631 ∩ dsx^FLP neurons.

A detailed characterization of NP2631 ∩ dsx^FLP neurons.

Behaviors of target flies remain largely unaffected by the optogenetic stimulation of NP2631 ∩ dsx^FLP neurons in tester flies.

dsx specifies the sexual dimorphism of NP2631 ∩ dsx^FLP neurons.

Heterozygosity of the fru^4-40 allele leaves the neuroanatomy and function of NP2631 ∩ dsx^FLP neurons largely unaltered.

NP2631 ∩ dsx^FLP neurons in fruF males promote courtship, but not aggressive, behavior.

Rasters of behaviors induced by the optogenetic activation of NP2631 ∩ dsx^FLP neurons in fruF males.

P1^a neurons are distinct from NP2631 ∩ dsx^FLP neurons.

Detailed characterization of P1^a neurons.

Detailed characterization of P1^a ∩ fru^FLP neurons.

P1^a and NP2631 ∩ fru^FLP neurons do not overlap.

Both dsx and fru specify the sexual dimorphism of P1^a neurons.

Heterozygosity of the fru^4-40 allele leaves the neuroanatomy and function of P1^a neurons largely unaltered.

P1^a neurons in fruF males are defective in promoting both courtship and aggressive behavior.

Videos

Representative behavior of a male tester fly that expresses.

Representative behavior of a male tester fly that expresses.

3D-rendered average image of NP2631 ∩ dsx^FLP neurons in male (green) and in female (magenta).

3D-rendered average image of NP2631 ∩ dsx^FLP neurons in male (green; duplication from Video 3) and in fruF male (yellow).

3D-rendered average image of NP2631 ∩ dsx^FLP neurons in female (magenta, duplication from Video 3) and in fruM female (blue).

Representative behavior of a fruF male tester fly that expresses.

3D-rendered average image of P1^a neurons (cyan) and NP2631 ∩ dsx^FLP neurons (orange) in male.

3D-rendered average image of P1^a neurons in male (green) and in fruF male (yellow).

Tables

Complete genotypes of Drosophila strains used in this study.

Additional files

Source data 1

Supplementary file 1

Transparent reporting form

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Sex of the target fly influences behaviors triggered by the optogenetic activation of social behavior-promoting neurons.

Additional characterization of the dsxGAL4 ∩ fruFLP neurons.

Performance of automated behavior classifiers.

Behaviors induced by optogenetic stimulation of dsxGAL4 ∩ fruFLP neurons across various LED frequencies.

NP2631 ∩ dsxFLP neurons that express both dsx and fru can promote courtship and aggressive behaviors.

Detailed description of behaviors induced by the optogenetic activation of NP2631 ∩ dsxFLP neurons.

A detailed characterization of NP2631 ∩ dsxFLP neurons.

Behaviors of target flies remain largely unaffected by the optogenetic stimulation of NP2631 ∩ dsxFLP neurons in tester flies.

dsx specifies the sexual dimorphism of NP2631 ∩ dsxFLP neurons.

Heterozygosity of the fru4-40 allele leaves the neuroanatomy and function of NP2631 ∩ dsxFLP neurons largely unaltered.

NP2631 ∩ dsxFLP neurons in fruF males promote courtship, but not aggressive, behavior.

Rasters of behaviors induced by the optogenetic activation of NP2631 ∩ dsxFLP neurons in fruF males.

P1a neurons are distinct from NP2631 ∩ dsxFLP neurons.

Detailed characterization of P1a neurons.

Detailed characterization of P1a ∩ fruFLP neurons.

P1a and NP2631 ∩ fruFLP neurons do not overlap.

Both dsx and fru specify the sexual dimorphism of P1a neurons.

Heterozygosity of the fru4-40 allele leaves the neuroanatomy and function of P1a neurons largely unaltered.

P1a neurons in fruF males are defective in promoting both courtship and aggressive behavior.

Representative behavior of a male tester fly that expresses.

Representative behavior of a male tester fly that expresses.

3D-rendered average image of NP2631 ∩ dsxFLP neurons in male (green) and in female (magenta).

3D-rendered average image of NP2631 ∩ dsxFLP neurons in male (green; duplication from Video 3) and in fruF male (yellow).

3D-rendered average image of NP2631 ∩ dsxFLP neurons in female (magenta, duplication from Video 3) and in fruM female (blue).

Representative behavior of a fruF male tester fly that expresses.

3D-rendered average image of P1a neurons (cyan) and NP2631 ∩ dsxFLP neurons (orange) in male.

3D-rendered average image of P1a neurons in male (green) and in fruF male (yellow).

Complete genotypes of Drosophila strains used in this study.

Source data 1

Supplementary file 1

Transparent reporting form

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

Additional characterization of the dsx^GAL4 ∩ fru^FLP neurons.

Behaviors induced by optogenetic stimulation of dsx^GAL4 ∩ fru^FLP neurons across various LED frequencies.

NP2631 ∩ dsx^FLP neurons that express both dsx and fru can promote courtship and aggressive behaviors.

Detailed description of behaviors induced by the optogenetic activation of NP2631 ∩ dsx^FLP neurons.

A detailed characterization of NP2631 ∩ dsx^FLP neurons.

Behaviors of target flies remain largely unaffected by the optogenetic stimulation of NP2631 ∩ dsx^FLP neurons in tester flies.

dsx specifies the sexual dimorphism of NP2631 ∩ dsx^FLP neurons.

Heterozygosity of the fru^4-40 allele leaves the neuroanatomy and function of NP2631 ∩ dsx^FLP neurons largely unaltered.

NP2631 ∩ dsx^FLP neurons in fruF males promote courtship, but not aggressive, behavior.

Rasters of behaviors induced by the optogenetic activation of NP2631 ∩ dsx^FLP neurons in fruF males.

P1^a neurons are distinct from NP2631 ∩ dsx^FLP neurons.

Detailed characterization of P1^a neurons.

Detailed characterization of P1^a ∩ fru^FLP neurons.

P1^a and NP2631 ∩ fru^FLP neurons do not overlap.

Both dsx and fru specify the sexual dimorphism of P1^a neurons.

Heterozygosity of the fru^4-40 allele leaves the neuroanatomy and function of P1^a neurons largely unaltered.

P1^a neurons in fruF males are defective in promoting both courtship and aggressive behavior.

3D-rendered average image of NP2631 ∩ dsx^FLP neurons in male (green) and in female (magenta).

3D-rendered average image of NP2631 ∩ dsx^FLP neurons in male (green; duplication from Video 3) and in fruF male (yellow).

3D-rendered average image of NP2631 ∩ dsx^FLP neurons in female (magenta, duplication from Video 3) and in fruM female (blue).

3D-rendered average image of P1^a neurons (cyan) and NP2631 ∩ dsx^FLP neurons (orange) in male.

3D-rendered average image of P1^a neurons in male (green) and in fruF male (yellow).