Fig. 4: Sensory specialization of Fru⁺ and Dsx⁺ P1 subpopulations in D. yakuba.

a,b, Left, template brain registrations of neurons labelled by intersection of Fru (a; Fru∩P1) or Dsx (b; Dsx∩P1) and the P1-driver 71G01 (Methods). Middle, averaged functional responses (∆F/F₀) aligned to tap of indicated female and average peak response (∆F/F₀) per male for a given female target. Right, average courtship bout length, total percentage time pursuing or total percentage time extending a unilateral wing towards a D. melanogaster female target before (Pre) or during (Stim) optogenetic stimulation of Fru∩P1>CsChrimson or Dsx∩P1>CsChrimson males and the fraction of flies engaging in these behaviours (sample sizes: Fru∩P1>GCaMP n = 6; Fru∩P1>CsChrimson n = 20; Dsx∩P1>GCaMP n = 10; Dsx∩P1>CsChrimson n = 20). c,d,e, Functional responses (as in a,b) of all Fru⁺ (c) or Dsx⁺ (d,e) neurons innervating the LPC in wild-type (c, n = 5; d, n = 8) or ∆ppk25 mutant (e, n = 8) males. f, Functional responses (as in b) of Dsx∩P1 to D. melanogaster mock-perfumed oe⁻ (oe⁻), 7,11-HD-perfumed (oe⁻ +7,11-HD), 7-T-perfumed (oe⁻ +7-T) or cVa-perfumed (oe⁻ +cVA). g, Courtship in the dark as captured by IFD of a D. yakuba male towards a conspecific female with constitutively silenced P1 subsets (Fru∩P1>Kir (middle), Dsx∩P1>Kir (bottom) or genetic control (top; 71G01-DBD; UAS-Kir)) (n = 6). Left, heatmaps for six pairs, aligned to courtship initiation (red, dotted line). Middle, histograms of time as a function of IFD for the same six courting pairs. Right, average courtship bout length in the 10 min following courtship initiation of each genotype (n = 25 each). For functional imaging (a–f), shading represents mean ± s.e.m. Points are individual males; error bars are mean ± s.e.m. For behavioural tests (a,b,e), points are individual males and bars are median. Statistics: ANOVA with Tukey’s post hoc (a–f), Wilcoxon test (a,b), Kruskal–Wallis test (g). Letters denote statistically different groups (P < 0.05). ****P < 0.0001.

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