Extended Data Fig. 7: Knockout of glucocorticoid receptors in specific brain regions in ABX-treated and SPF mice.
From: Microbiota regulate social behaviour via stress response neurons in the brain
a, d, g Diagrams of virus injection into the DG (Nr3c1ΔDG; relevant to Fig. 2h, i), BNST (Nr3c1ΔBNST; relevant to Fig. 2j, k) and hypothalamus (Nr3c1ΔHYPO; relevant to Fig. 2l, m). b, e, h, Non-social activity recorded using the RSI paradigm in ABX mice (subject), in the context of SPF novel mice. b, No difference in non-social activity was detected in ABX Nr3c1ΔDG mice. Control vs Nr3c1ΔDG P = 0.1191. n = 7 mice per group (subject only). e, Minimal difference in non-social activity was detected in ABX Nr3c1ΔBNST mice. A decrease in non-social activity was detected only at the second CMC injection (*P = 0.0363). n = 7 mice per group (subject only). h, No difference in non-social activity was detected in Nr3c1ΔHYPO mice. Control vs Nr3c1ΔHYPO P = 0.0652. n = 6 control, 5 Nr3c1HYPO mice per group (subject only). c, Quantification of c-Fos+ cells in various brain regions of ABX Nr3c1ΔDG and control mice. Decreased c-Fos+ cells were detected in the PVN and adBNST after social interaction in ABX Nr3c1ΔDG mice compared to control mice. There was no change in c-Fos expression in the DG of ABX Nr3c1ΔDG mice. PVN ****P < 0.0001, adBNST **P = 0.0022, DG P = 0.9714. n = 7 mice per group. f, Quantification of c-Fos+ cells in various brain regions of ABX Nr3c1ΔBNST and control mice. Decreased c-Fos+ cells were detected in the PVN and adBNST after social interaction in ABX Nr3c1ΔBNST mice compared to control mice. There was no change in c-Fos staining in the DG. PVN ****P < 0.0001, adBNST ***P = 0.0002, DG P = 0.4187. n = 7 mice per group. i, Quantification of c-Fos+ cells in various brain regions of ABX Nr3c1ΔHYPO and control mice. Increased c-Fos+ cells were detected in DG and adBNST after social interaction in ABX Nr3c1ΔHYPO mice compared to control mice. There was no change in c-Fos expression in the PVN. PVN P = 0.1163, adBNST P = 0.0688, DG *P = 0.0389. n = 6 control, 5 Nr3c1ΔHYPO mice per group. j, Neuronal activity was measured by c-Fos staining of various brain sections. Representative images of c-Fos staining after social interaction in PVN, adBNST, and DG in Nr3c1ΔDG (from 7 animals each group), Nr3c1ΔBNST (from 7 animals each group) and Nr3c1ΔHYPO mice (from 6 control and 5 Nr3c1ΔHYPO animals), and their corresponding control groups. Scale bars, 100 μm. k, l, Social activity (k) and non-social activity (l) were tested using the RSI paradigm in SPF test mice (subject), in the context of SPF novel mice. SPF Nr3c1ΔDG mice did not display altered social activity (P = 0.4323) or non-social activity (P = 0.8234) compared to control mice with vehicle injection. n = 5 mice per group (subject only). m, Serum corticosterone concentrations show no change after social interaction in SPF Nr3c1ΔDG mice. P = 0.9935. n = 5 mice per group. n, o, Social activity (n) and non-social activity (o) were tested using the RSI paradigm in SPF test mice (subject) in the context of SPF novel mice. SPF Nr3c1ΔBNST mice showed reduced social activity compared to control mice with vehicle injection (*P = 0.0473). No change was observed in non-social activity between SPF control and Nr3c1ΔBNST mice (P = 0.4742). n = 5 mice per group (subject only). p, Serum corticosterone concentrations show no change after social interaction in SPF Nr3c1ΔBNST mice. P = 0.9312. n = 5 mice per group. q, r, Social activity (q) and non-social activity (r) were tested using the RSI paradigm in SPF test mice (subject), in the context of SPF novel mice. SPF Nr3c1HYPO mice did not display altered social activity (P = 0.1823) or non-social activity (P = 0.2339) compared to control mice with vehicle injection. n = 5 mice per group (subject only). s, Serum corticosterone concentrations show no change after social interaction in SPF Nr3c1ΔHYPO mice. P = 0.0701. n = 5 mice per group. Data shown as individual points with mean ± s.e.m. Data analysed by two-way ANOVA, repeated measures (b, e) with mixed effect (h) with Bonferroni’s multiple comparison post hoc test; two-tailed unpaired t-test (c, f, i, k–s). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ND: no difference. For more statistical details, see Supplementary Information.
