Extended Data Fig. 5: VMHvlMC4R neurons function specifically to drive physical activity.
From: Oestrogen engages brain MC4R signalling to drive physical activity in female mice
a, Distance travelled over time in female and male mice following a single injection of CNO (repeated-measures two-way ANOVA interaction effect female: F39,312 = 11.96, P < 0.0001; male: F39,312 = 6.898, P < 0.0001). Holm–Šidák multiple comparisons test. b, Total time spent immobile in intact female and male VMHvlCre− controls (n = 5/4) and VMHvlMC4R::hM3Dq mice (n = 5/5) (repeated-measures two-way ANOVA female interaction effect F1,8 = 33.89, P = 0.0004, post hoc P < 0.0001 and male interaction effect F1,8 = 96.79, P = 0.0005, post hoc P < 0.0001). Holm–Šidák multiple comparisons test. c, Thermal imaging of BAT surface temperatures for VMHvlCre− (left mouse) and VMHvlMC4R:: hM3Dq (right mouse) female mice treated with CL-316,243. d, No differences were observed in Ucp1 mRNA in the BAT from VMHvlCre− and VMHvlMC4R::hM3Dq mice collected 1.5 h after a single CNO injection. e, Body weights for female VMHvlCre− (n = 5) and VMHvlMC4R::hM3Dq (n = 6, baseline measurement includes 1 mouse with mistargeted injection) mice prior to glucose tolerance test (GTT). f, GTT glucose levels for intact female cohorts treated with saline or CNO and total area under the curve (AUC) (unpaired two-tailed Student's t-test t8 = 2.824, P = 0.0223). g, Body weight normalized food consumption in female mice (n = 5/5) following Sal/CNO injection during light dark period (ZT12–ZT17) (repeated-measures two-way ANOVA Dark period interaction effect F1,8 = 3.502, P = 0.0982, post hoc P = 0.0489). Holm–Šidák multiple comparisons test. h, Sustained physical activity increase across light/dark periods in VMHvlMC4R::hM3Dq (n = 5) female mice administered CNO-H2O as compared to VMHvlCre− female mice (n = 5) or during exposure to plain drinking water (H2O). i, j, Cumulative distance travelled (i) and number of rearing events (j) during light/dark periods following administration of CNO or water during the light stage (i repeated-measures two-way ANOVA F1,8 = 15.8, P = 0.0041, post hoc P = 0.0006 and j, repeated-measures two-way ANOVA F1,8 = 15.8, P = 0.0041, post hoc P = 0.0006). Holm–Šidák multiple comparisons test. k, Starting body weights and weight change during continuous administration of CNO-H2O for intact female mice. l, ERα and mCherry expression in the VMHvl following targeted injection of Cre-dependent AAV-hM4Di-mCherry into a female Mc4r-t2a-cre mouse. m, Number of rearing episodes during the dark period in VMHvlMC4R::hM4Di (n = 8) and VMHvlCre− (n = 4) intact female mice following administration of plain or DCZ-laden drinking water. Data are mean ± s.e.m. or box plots (described in Fig. 1c legend).
