Fig. 1: Deep learning analysis of natural behaviour reveals how pain and opioids shape internal affective–motivational states.
a, Schematic of the standardized LUPE chamber. b, A 20-body point DLC pose-tracking model was built from male and female mouse pain behaviour. c, Behaviour segmentation models trained iteratively on supervised annotations of behaviours with A-SOiD, followed by unsupervised sub-clustering with B-SOiD. d, Motion energy heat maps illustrating spatial trajectories and intensity distributions for the six primary behavioural repertoires. e, Temporal probability plots for the six primary behaviour repertoires in 1-min bins, comparing uninjured mice to mice with left hindpaw injections of 1% formalin, 5% formalin or capsaicin. f, Raster plots of behaviour transitions within a 30-s window. g, Procedure for behavioural state inference from statistical structure of spontaneous behaviour. h, Left, model centroid transition matrices characterizing each of six inferred states. Right, comparing the fraction occupancy of mice in each state between uninjured (grey), formalin (magenta) and capsaicin (cyan) pain models (n = 20 per group; one-way analysis of variance (ANOVA); Tukey correction: Pstate1 = 0.0007, Pstate3 = 0.0078 and Pstate4 < 0.0001). i, Two-dimensional (2D) visualization of PCA of state occupancies across pain models. j, Magnitude of coefficients of each state in each PCA. k, Scores of each animal along PC1 (top) and PC2 (bottom) across pain models (n = 20 per group; one-way ANOVA; Tukey correction: PPC1 = 0.0027 and PPC2 = 0.0082). l, Dose–response of morphine on PC1 (top) and PC2 (bottom) scores in uninjured, formalin-administered and capsaicin-administered mice (n = 20 per group and dose; one-way ANOVA; Tukey correction: PPC1 uninjured < 0.0001, PPC1 formalin < 0.0001, PPC1 capsaicin < 0.0001, PPC2 uninjured < 0.0001, PPC2 formalin < 0.0001 and PPC2 capsaicin < 0.0001). ⋆P < 0.05. Bars are mean; dots are individual animals; vertical lines and shaded areas are s.e.m. See Supplementary Table 1 (rows 1–14) for statistics. NS, non-significant. Scale bar, 2.5 s (f).
