Fig. 1: lPBN Y1 receptor-expressing neurons influence sustained and chronic pain.

a, Mice were injected with formalin in the hindpaw before sample collection. PBN sections were analysed using a spatial molecular imager (CosMx). n = 3 mice, 4 sections each. ROI, region of interest. b, Uniform manifold approximation and projection (UMAP) embedding of PBN neuron clusters. c, Representative spatial map of molecularly defined clusters. D, dorsal; L, lateral; M, medial; V, ventral. d, UMAP embeddings and representative spatial maps showing normalized expression levels of Calca and Penk. UMAP scale maximum: Calca, 3.7; Penk, 4.5. Spatial scale maximum: Calca, 3.3; Penk, 4.2. e, Normalized expression levels of Tacr1 and Npy1r. UMAP scale maximum: Tacr1, 1.9; Npy1r, 2.25. Spatial scale maximum: Tacr1, 1.4; Npy1r, 1.5. f, Normalized expression levels of Fos. UMAP scale maximum: 2.55. Spatial scale maximum: 1.75. g, Percentage of Npy1r-expressing and non-expressing cells co-expressing Fos. Box plots show the median (centre line), 25th–75th percentiles (box) and 5th–95th percentiles (whiskers) (n = 6 to 12 sections per group; two-way ANOVA, main effect of group P < 0.001). h, Assays used to assess pain behaviour. i, Strategy to inhibit lPBN Y1R neurons. j,k, Time licking paw after formalin during phase 1 (j) and phase 2 (k) in control or with Y1R neuron inhibition (n = 11 control, n = 10 hm4D(Gi) mice; unpaired two-sided t-test, not significant (j), P = 0.0352 (k)). l,m, Withdrawal threshold (mechanical; l) or duration (acetone; m) in hM4D(Gi)-expressing mice before SNI and after SNI with a vehicle (Veh) or clozapine-N-oxide (CNO) injection (n = 8 mice; one-way ANOVA, P < 0.001). n, Strategy to activate lPBN Y1R neurons. o,p, Withdrawal threshold (mechanical; o) or duration (acetone; p) in hM3D(Gq)-expressing mice after a vehicle or CNO injection (n = 7 mice; paired two-sided t-test, P = 0.0005). q, Time spent in the centre of an open field (n = 8 mice; two-sided Wilcoxon matched pairs signed-rank test, P = 0.0078). r, Percentage of time spent in the side of a chamber paired with CNO (n = 6 control, n = 7 hm4D(Gi) mice; two-way ANOVA, main effect of group P = 0.0002, group × time interaction P < 0.001). Data are mean ± s.e.m. unless noted otherwise. Grey dots and lines represent individual mice. t-test and post hoc comparisons: *P < 0.05, **P < 0.01, ***P < 0.001. ANOVA main effect of group: ^#P < 0.05, ^##P < 0.01, ^###P < 0.001. ANOVA interaction: ^†P < 0.05, ^††P < 0.01, ^†††P < 0.001.

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