Extended Data Fig. 8. MPOA-projecting MeA^Tac1∩Vgat neurons drive affiliative allogrooming in males and females.

a, b, Example images showing axonal terminals of MeA^Tac1∩Vgat neurons in the MPOA in male (a) and female (b) animals, revealed by immunostaining for EYFP in Tac1^Cre/+/Vgat^Flp/+ animals injected with Con/Fon-ChR2-EYFP. Scale bar, 200 μm. (c, h) Schematics of viral injection and fiber implantation strategies for soma stimulation of retrogradely labeled, MPOA-projecting MeA^Tac1∩Vgat neurons (c) or stimulation of the axonal projection of MeA^Tac1∩Vgat neurons in the MPOA (h). d–g, Duration (d, f) and onset latency (e, g) of allogrooming during photostimulations in EYFP control and ChR2 males (d, e) and females (f, g) with soma stimulation of MPOA-projecting MeA^Tac1∩Vgat neurons. i–l, Duration (i, k) and onset latency (j, l) of allogrooming during photostimulations in EYFP control and ChR2 males (i, j) and females (k, l) with stimulation of the MPOA projection of MeA^Tac1∩Vgat neurons. m–o, Probability of allogrooming toward stressed partners (fraction of trials showing allogrooming at different time points) with respect to stimulation onset (m, n) and allogrooming duration (o) during photostimulations of the MPOA projection of MeA^Tac1∩Vgat neurons without or with infusion of lidocaine in the MeA. Blue areas: duration of light illumination; time 0: stimulation onset. p, Example image showing axonal projections of MeA^Tac1∩Vgat neurons in the PMv. Scale bar, 200 μm. q, Duration of allogrooming during photostimulation of the MPOA or PMv projection of MeA^Tac1∩Vgat neurons. Data for MPOA projection stimulation are the same as those in Fig. 4r and are presented here for comparison. Boxplots: median with quartiles, 1.5 × IQR and outliers. d–g, i–l, EYFP control, MeA soma stimulation, n = 52 trials in 4 females and 29 trials in 2 males. ChR2, MeA soma stimulation, n = 68 trials in 4 females and 59 trials in 2 males. EYFP control, MPOA projection stimulation, n = 45 trials in 4 females and 22 trials in 2 males. ChR2, MPOA projection stimulation, n = 40 trials in 3 females and 53 trials in 3 males. m–o, n = 36 trials in 4 mice (24 trials from 3 females and 12 trials from 1 male) for the “no lidocaine” group. n = 58 trials in 4 mice (39 trials from 3 females and 19 trials from 1 male) for the “lidocaine” group. q, n = 93 trials in 6 mice (40 trials in 3 females and 53 trials in 3 males) for MPOA stimulation, n = 118 trials in 4 mice (65 trials in 2 females and 53 trials in 2 males) for PMv stimulation. Two-sided Wilcoxon rank-sum test. ***P < 0.001. ns, not significant. For details of statistical analyses, see Supplementary Table 1.