Extended Data Fig. 10: Characterizations of TRAP⁺ neurons in the MPOA.

a-g, Characterizations of TRAP⁺ neurons. a1, b1, c1, d1, Example images showing hM4Di-mCherry expression in pup-TRAP⁺ neurons (a1), non-social-TRAP⁺ neurons (b1), stressed-TRAP⁺ neurons (c1), and unstressed-TRAP⁺ neurons (d1). Scale bars, 500 μm. a2, b2, Example images showing pup-TRAP⁺ neurons (a2) or non-social-TRAP⁺ neurons (b2) expressing hM4Di-mCherry (red), cells expressing Fos (green) following pup interactions, and the overlap between the two populations in the MPOA. Scale bar, 50 μm. c2, d2, Example images showing stressed-TRAP⁺ neurons (c2) or unstressed-TRAP⁺ neurons (d2) expressing hM4Di-mCherry (red), cells expressing Fos (green) following interactions with stressed adults, and the overlap between the two populations in the MPOA. Scale bar, 50 μm. e, Percentages of pup-TRAP⁺ and non-social-TRAP⁺ neurons that overlapped with Fos⁺ cells. f, Percentages of stressed-TRAP⁺ and unstressed-TRAP⁺ neurons that overlapped with Fos⁺ cells. g, Percentage of hM4Di-expressing neurons within the NeuN⁺ neurons in the MPOA of TRAP animals. h, i, j, k, Percentages of time that pup-TRAP animals (h), non-social-TRAP animals (i), stressed-TRAP animals (j), and unstressed-TRAP animals (k) engaged in self-grooming behavior during periods when they did not display social behavior (social investigation or allogrooming) in the prosocial interaction assay following saline or CNO injection. l, o, r, u, Example locomotion trajectories of pup-TRAP animals (l), non-social-TRAP animals (o), stressed-TRAP animals (r), and unstressed-TRAP animals (u) in the open field test following saline (control) or CNO injection. m, n, p, q, s, t, v, w, Time spent in the centers and travel distance of pup-TRAP animals (m, n), non-social-TRAP animals (p, q), stressed-TRAP animals (s, t), and unstressed-TRAP animals (v, w) in the open field test following saline (control) or CNO injection. (e-k, m, n, s, t, p, q, v, w), The center line in the boxplots indicates the median, the box limits indicate the upper and lower quartiles, and the whiskers indicate the range from the minimum to the maximum value. (e, f), Two-sided Wilcoxon rank-sum test. (g), Kruskal-Wallis test with post hoc Dunn’s multiple comparison test. (h-k, m, n, s, t, p, q, v, w), Two-sided Wilcoxon signed-rank test. n = 8 independently injected hemispheres (3 brain sections per hemisphere) in pup-TRAP group and 6 independently injected hemispheres (3 brain sections per hemisphere) in non-social-TRAP group in (e). n = 8 independently injected hemispheres (3 brain sections per hemisphere) in each group in (f, g). n = 13 mice in (h, m, n), 12 mice in (i, p, q), 20 mice in (j), 19 mice in (s, t), 14 mice in (k, v, w). ns, not significant. See Supplementary Table 1 for details of statistical analyses. *P < 0.05. **P < 0.01. ***P < 0.001. ns, not significant.

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