Extended Data Fig. 1: Chemogenetic and optogenetic silencing of BNSTpr^Esr1 neurons disrupts aggression and mating.

a, Illustration of hM4d (Gi)-mediated silencing in bilateral BNSTpr^Esr1 neurons. b, Example behavior raster plot from 1 animal across 4 recording sessions. c, Percent of BNSTpr^Esr1 neurons that express mCherry. Values are plotted as mean ± SEM, n = 6 mice. d–m, Measurements performed before (pre-CNO: saline injection, gray points) vs. after (CNO injection, maroon points) chemogenetic silencing of BNSTpr, of time spent sniffing male intruders (d), number of attack bouts toward male intruders (e), time spent attacking male intruders (f), time spent sniffing female intruders (g), number of mount bouts toward female intruders (h), time spent mounting female intruders (i), time spent mounting male intruders (j), time spent attacking female (k), and time spent attacking male (l), time spent mounting female (m) in mCherry/CNO only controls (l, m) per 30-minute session. 100µl containing either saline (pre-CNO) or 7.5mg/kg CNO was given i.p. 60 min prior to start of behavior tests. n, Illustration of bilateral optogenetic silencing in BNSTpr. o, t, Mean probability of sniff behavior occurring relative to onset of optogenetic silencing. p, u, Duration of sniffing toward male (p) or female (u) intruders. q, v, Duration of attack (q) toward male intruders or mount (v) toward female intruders as a percentage of the duration of optogenetic silencing. “Sham” controls were the same animals during a 10s “sham stimulation,” i.e., without light. r, w, Percent of time spent interacting with male or female urine (r), or male or female conspecifics separated by a meshed barrier (w) before and during (r) or before, during and after (w) optogenetic silencing of BNSTpr. Optogenetic silencing conditions: 470nm @ ~1 mW/mm² for 10 s (o, p, q, t, u, v). 470nm @ ~1 mW/mm² for 10 min (r, s, w, x) Statistics: For Chemogenetic inhibition, two-sided Wilcoxon signed-rank test (d–m). Values are plotted as total time or total bouts per 30-minute session per animal. n = 7 mice (d–k), n = 4 mice (l, m). For Optogenetic inhibition during interactions with male or female conspecifics (o, p, q, t, u, v), two-sided Kolmogorov-Smirnov test (o, t), two-sided Wilcoxon signed-rank test (p, q, u, v). Male-male trials (o, p, q), n = 131 stim and 125 sham trials pooled from 10 mice. Male-female trials mice (t, u, v), n = 181 stim and 170 sham trials pooled from 10 mice. For Optogenetic inhibition during urine preference (r) and “pencil cup” (mesh barrier) (w) tests, two-way ANOVA with Bonferroni correction, n = 6 mice. Values are plotted as mean ± SEM. ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05. The mouse brain images in this figure (a, n) have been reproduced from ref. ⁴¹.