Extended Data Fig. 8: Molecular and topographical distinctions of DCN neurons that respond to food intake.

(a-f) Expression of activity-regulated transcript Homer1a⁶³ (red) in the three major cerebellar nuclei following food deprivation (a, c, e) or refeeding (b, d, f). (g-l) Spp1 expression (green) in vGluT2⁺ neurons (blue) (g, i, k), and colocalised Spp1 (cyan, Spp1⁺vGluT2⁺ neurons) (h, j, l) in the three major cerebellar nuclei. (m-r) Celf4 expression (blue) in vGluT2⁺ neurons (red) (m, o, q), and colocalised Celf4 (magenta, Celf4⁺vGluT2⁺ neurons) (n, p, r) in the three major cerebellar nuclei. Scale bar, 100 µm. (s) Summary of the expression of Spp1 and Celf4 and the distribution of Homer1a+ DCN neurons. (t) Schematic of fibre photometry system. (u-w) Fibre targeting aDCN-LAT glutamatergic neurons in vGluT2::Cre mouse (u), expression of GCaMP6s (green) and vGluT2 (red) (v, w). Scale bar, 20 µm. (x-z) Heatmaps depicting ∆F/F of GCaMP6 signals in the aDCN-LAT glutamatergic neurons of ad libitum fed (x) and food-deprived (y) mice response to chow, and ad libitum fed mice response to non-food object (z, marble). Signals are aligned to the introduction of chow or non-food object (red line) (n = 7 mice). (aa) Average ∆F/F of GCaMP6 signals in the aDCN-LAT glutamatergic neurons (490 nm, green, and control 405 nm, magenta). Signals are aligned to the introduction of non-food object (red line). Dark line represents the mean and lighter shaded area represents SEMs (n = 7). (bb-cc) Mean (bb) and max (cc) ∆F/F GCaMP6s signals of aDCN-LAT glutamatergic neurons in response to chow, in ad libitum fed (grey) and food-deprived (red) mice, and response to non-food object in ad libitum fed mice (n=7, one-way ANOVA (bb) P < 0.001, (cc) P < 0.001; Holm-Sidak’s, (bb) P < 0.006, P = 0.475, P < 0.003, (cc) P<0.009, P=0.651, P = 0.011, ad libitum fed chow versus food deprivation chow, ad libitum fed chow versus non-food, food deprivation chow versus non-food, respectively). Data are expressed as mean ± SEM, two-sided P values, post-hoc comparisons: *P < 0.05, **P < 0.01. Statistical analysis in Supplementary Table 1

Source data.