Extended Data Fig. 9: Retrograde neural tracing of the PVN and adBNST of GF and ABX-treated mice.
From: Microbiota regulate social behaviour via stress response neurons in the brain
a, CTB-488 was stereotaxically injected into the PVN of GF and SPF C57BL/6J mice. The brains were removed seven days after injection. CTB-488 retrograde tracing was visualized by confocal imaging of brain sections counterstained with an antibody against NeuN. GF mice were treated with ABX after surgery. b, Quantification of CTB-488+ cells in various brain regions of SPF and GF mice. adBNST (P = 0.229); LS (P = 0.1373); MeA (P = 0.358). No difference in CTB-488 retrograde-labelled cells was detected between SPF and GF mice. n = 4 mice per group. c, Representative images (from 4 animals each group) of CTB-488 (green) labelling and NeuN (magenta) staining in the adBNST, MeA, and LS of SPF and GF mice. Scale bars, 100 μm. d, CTB-488 and Fluorogold were stereotaxically injected into the PVN and adBNST, respectively, in vehicle and ABX Crh-ires-Cre;Ai14D mice. Brains were removed 7 days after injection. Retrograde tracers were visualized by confocal imaging of brain sections counterstained with antibodies. e, Quantification of CTB-488+ cells in various brain regions of vehicle and ABX mice. No difference in CTB-488 retrograde-labelled cells was detected between vehicle and ABX mice. adBNST P = 0.2064, LS P = 0.4332, MeA P = 0.4366. n = 4 mice per group. f, Quantification of Fluorogold (FG)+ cells in various brain regions of vehicle and ABX mice. No difference in Fluorogold retrograde-labelled cells was detected between vehicle and ABX mice. adBNST P = 0.8922, LS P = 0.8715, MeA P = 0.7284. n = 4 mice per group. g, Representative images (from 4 animals each group) of CTB-488 (green) labelling, Fluorogold (blue) staining, and CRH+ tdTomato (red) cells in the adBNST, MeA, LS, and PVN of vehicle and ABX mice. Scale bars, 100 μm. Data shown as individual points with mean ± s.e.m. Data analysed by two-tailed unpaired t-test (b, e, f). ND: no difference. For more statistical details, see Supplementary Information.
