Extended Data Fig. 4: aVMHvlOXTR cells increase responses to the aggressor after defeat in male mice.
From: A dedicated hypothalamic oxytocin circuit controls aversive social learning
a. Schematics of virus injection and a representative histology image. Dashed line marks the aVMH. Scale bar: 200 µm. Brain illustration in is based on a reference atlas from https://atlas.brain-map.org/. b. Cartoon illustration of the RI test. c-e. Representative Z-scored GCaMP6f traces during RI tests with a BC male intruder (c), a SW male intruder (d), or resident SW (e). c2, d2 and e2 show the enlarged boxed areas. f. Post-event histograms (PETHs) of GCaMP6f signal aligned to initial opponent encounters, only including sessions without defeat or attack during the first 10 s. g. Peak GCaMP6f response within the first 10 s of RI tests. h-i. PETHs of GCaMP6f signals aligned to close investigation (h) and agonistic interactions (i). j. Averaged Z-scored responses during various social behaviors. k. Experimental timeline. l. Heatmaps showing the body center location of a representative test male during pre- and post-defeat SI tests. m. Distribution of the distance between the test animal’s body center and cup center during pre- and post-defeat SI tests. n. The percentage of total time test mice spent around the aggressor cup during pre- and post-defeat SI tests. o. The percentage of total time the test mice spent investigating the constrained aggressor during pre- and post-defeat SI tests. p. Representative Z-scored GCaMP6f traces from a recording male mouse during pre-defeat (p1) and post-defeat (p2) SI tests. Shades mark investigation events. q. PETHs of Z-scored GCaMP6f signals aligned to the onset of investigating aggressor during pre- and post-defeat SI tests. r. Average Z-scored ΔF/F of aVMHvlOXTR cells during investigating aggressor in pre- and post-defeat SI tests. s. Scatter plot showing the correlation between investigation time change index and change in Z-scored GCaMP response to the aggressor during post-defeat SI tests from the pre-defeat level. t. Representative Z-scored GCaMP trace (black) overlaid with the velocity trace (orange) during the post-defeat SI test. Blue indicates the period when the test animal quickly retreated from the aggressor. u. PETHs of Z-scored GCaMP signal (black) and velocity (orange) aligned to the retreat onset. v. The retreat onset GCaMP signal is significantly higher than the retreat offset signal. w. Representative Z-scored GCaMP trace (black) overlaid with the velocity trace (orange) during the post-defeat SI test. Gray indicates when the test animal stayed immobile and far from the aggressor. x. PETHs of Z-scored GCaMP signal (black) and velocity (orange) aligned to immobility onset. Immobility trials are defined as velocity <1 pixel/frame lasting for > 0.5 s. y. The mean GCaMP signal during immobility. Plots with shades and error bars represent mean±s.e.m. Lines and circles represent individual animals. Numbers on the plots indicate the number of animals. One-way ANOVA with Tukey’s multiple comparisons (g), Two-way repeated-measure ANOVA with Sidak’s multiple comparisons (j), Wilcoxon matched-pairs signed rank test (n and o), paired t-test (r, v), one-sample t-test (y), and Pearson cross-correlation (s). All statistical tests are two-tailed. *p < 0.05. See Supplementary Table 1 for detailed statistics.
