Extended Data Fig. 6: Chemogenetic inhibition of Npy1RPVH neurons does not affect energy expenditure.
From: Reciprocal activity of AgRP and POMC neurons governs coordinated control of feeding and metabolism
a, Scheme of AAV-flex-hM4DGi injection into PVH area of Npy1R-Cre+/− mice. b, Representative image of NPy1RPVH neurons that express hM4DGi-mCherry protein (red) showing almost no colocalization with Fos mRNA marker (blue) after CNO injection (3 mg/kg, 1h). c, Quantification of chemogenetic-induced silencing of mCherry-expressing Npy1R+ neurons in PVH area, and characterization of neurotransmitter nature by colocalization with Slc17a6+ or Slc32a1+ markers (n = 4 male mice). d, Cumulative food intake curves after vehicle or CNO injection in Control-GiPVH (Npy1R-Cre −/−) mice (n = 6 male mice) e, Food intake during the time intervals from Control-GiPVH Gi male mice (n = 6 male mice) treated with vehicle (open bars) or CNO (closed bars). f, Average RER during the light and dark cycles in Control-Gi and Npy1R-Gi mice (n = 6 and 9 male mice, respectively) treated with vehicle (open bars) or CNO (closed bars. g, Average of energy expenditure (EE) along the light and dark cycles in Control-Gi and Npy1R-Gi mice (n = 6 and 9 male mice, respectively) treated with vehicle (open bars) or CNO (closed bars). h, Average of activity counts during the light and dark cycles in Control-Gi and Npy1R-Gi mice (n = 6 and 9 male mice, respectively) treated with vehicle (open bars) or CNO (closed bars). i, Cumulative food intake curves after vehicle or CNO injection from female Npy1R-Gi mice treated with vehicle (open dots) or CNO (closed dots) (n = 6 female mice), *P = 0.0364 Veh. Vs. CNO. j, Food intake during the time intervals from Npy1R-Gi female mice (n = 6 female mice) treated with vehicle (open bars) or CNO (closed bars); *P = 0.102, **P = 0.0034 and ****P < 0.001 Veh vs. CNO. k, Average of respiratory coefficients (RER) for light and dark cycle from female Npy1R-Gi mice treated with vehicle (open bars) or CNO (closed bars) (n = 6 female mice). l, Average of energy expenditure (EE) for light and dark cycle from female Npy1R-Gi mice treated with vehicle (open bars) or CNO (closed bars) (n = 6 female mice). m, Average of activity counts during light and dark cycle from female Npy1R-Gi mice treated with vehicle (open bars) or CNO (closed bars). n, Scheme of the experimental design. o, Cumulative food intake along 3 d of chronic treatment with vehicle (grey lines) or CNO (red lines) in drinking water for Npy1R-Gi female mice (n = 12 mice) ****P < 0.0001. p, Percentage of body weight gain after 3 d with chronic treatment with vehicle or CNO in female Npy1R-Gi mice (n = 12 female mice) *P = 0.0102. q, Average of food intake during light and dark cycles along the 3-day chronic treatment with vehicle or CNO in drinking water (n = 12 female Npy1R-Gq mice) *P = 0.0251 and **P = 0.0019. r, Average respiratory coefficient (RER) from 3 d of chronic treatment during light and dark cycles in female Npy1R-Gi mice (n = 12 female mice). ****P < 0.0001. Data represent average ± s.e.m of the biological replicates. Boxplots indicate median ±min/max and include data points of individual mice entering the analysis. Statistical test: two tailed paired T-test for p, 2-way ANOVA for d – r; followed by Bonferroni’s test for I, k, o and r; and Sidak’s test for j, l, m and q. Source data and further details of statistical analyses are provided as a Source Data file. White scale bar = 100 µm in a, d. Black scale bar = 1500 µm in e. Created with BioRender.com.
