Extended Data Fig. 10: Activation of aDCN neurons robustly increases striatal dopamine signalling that correlates with reduced food intake.

(a) Schematic depicting hM3D(Gq) expression in the DCN combined with GRAB^DA expression⁴¹ and fibre implant in the ventral striatum which receives projections from the ventral tegmental area (VTA) dopamine (DA) neurons^64,65. (b) GRAB^DA expression and fibre placement in the ventral striatum. Scale bar, 1000 µm. (c, d) Average ∆F/F of GRAB^DA signals in the ventral striatum of food-deprived mCherry control (c) or aDCN-LAT hM3D(Gq) (d) mice treated with vehicle or CNO. Signals are aligned to the vehicle or CNO injection (red line). Dark line represents the mean and lighter shaded area represents SEMs. Corresponding heatmaps (right) depict ∆F/F of GRAB^DA signals in each mouse (n = 6 control mice, grey; n = 6 hM3D(Gq) aDCN-LAT mice, green). (e) Average ∆F/F of GRAB^DA signals in 3-min bins (n = 6 control mice, grey; n = 6 hM3D(Gq) aDCN-LAT mice, repeated measures two-way ANOVA interaction P < 0.001, main effect P < 0.001; Holm-Sidak’s, P = 0.027/ = 0.397/=0.625 (12-15 min), P < 0.001/<0.001/ < 0.001 (15-18 min), P < 0.001/<0.001/ < 0.001 (18-21 min), P<0.001/ < 0.001/ < 0.001 (21-24 min), P < 0.001/ < 0.001/ < 0.001 (24-27 min), P < 0.001/ < 0.001/ < 0.001 (27-30 min), hM3D(Gq) CNO to hM3D(Gq) vehicle/control CNO/control vehicle respectively). (f) Maximum ∆F/F of GRAB^DA signals in the ventral striatum of vehicle or CNO treated food-deprived mice with aDCN-LAT hM3D(Gq) (n = 6, paired t-test, P = 0.011). (g) Mean ∆F/F of GRAB^DA signals in the ventral striatum (n = 6, paired t-test, P = 0.002). (h) Maximum ∆F/F of GRAB^DA signals in the ventral striatum of vehicle or CNO treated food-deprived mice with aDCN mCherry control mice (n = 6, paired t-test, P = 0.242). (i) Mean ∆F/F of GRAB^DA signals in the ventral striatum of vehicle or CNO treated food-deprived mice with aDCN mCherry control mice (n = 6, paired t-test, P = 0.418). (j) Scatter plot comparing changes in GRAB^DA signals to amount of chow consumed in 1 h following activation of the aDCN in hM3D(Gq)-expressing mice treated with CNO (n = 13, Pearson correlation). (k) Average ∆F/F of GRAB^DA signals in the ventral striatum of food-deprived aDCN-INT hM3D(Gq) mice treated with vehicle or CNO. Signals are aligned to the vehicle or CNO injection (red line). Dark line represents the mean and lighter shaded area represents SEMs. Corresponding heatmaps (right) depict ∆F/F of GRAB^DA signals in each mouse (n = 7). (l) Average ∆F/F of GRAB^DA signals in 3-min bins following vehicle or CNO treatment of the aDCN-INT with hM3D(Gq) (n = 7, repeated measures two-way ANOVA interaction P = 0.301). (m) Maximum ∆F/F of GRAB^DA signals in the ventral striatum of vehicle or CNO treated food-deprived mice with aDCN-INT hM3D(Gq) mice (n = 7, paired t-test, P = 0.410). (n) Mean ∆F/F of GRAB^DA signals in the ventral striatum of vehicle or CNO treated food-deprived mice with aDCN-INT hM3D(Gq) mice (n = 7, paired t-test, P = 0.367). Data are expressed as mean ± SEM, two-sided P values, t-tests and post-hoc comparisons: *P < 0.05, **P < 0.01, ***P < 0.001; ANOVA interaction: ∞∞∞P < 0.001; ANOVA main effect of group: ¤¤¤P < 0.001. Statistical analysis in Supplementary Table 1

Source data.