Figure 3. Activation of PVHBDNF neurons increases energy expenditure. A, G, Locomotor activity over 12 h after sequential VEH and CNO injection in male and female Bdnf2A-Cre/+ mice injected with either AAV2-hSyn-DIO-mCherry (mCherry) or AAV2-hSyn-DIO-hM3D(Gq)-mCherry (hM3Dq) into the PVH. RM Two-way ANOVA between mCherry and hM3Dq groups: males (over 4 h post-VEH: F(1,8) = 0.1209 and p = 0.7371; over 4 h post-CNO: F(1,8) = 119.2 and p < 0.0001) and females (over 4 h post-VEH: F(1,21) = 0.3294 and p = 0.5721; over 4 h post-CNO: F(1,21) = 31.50 and p < 0.0001). B, H, Oxygen consumption (VO2) over 12 h after sequential VEH and CNO injection in male or female mCherry or hM3Dq mice. RM two-way ANOVA between mCherry and hM3Dq groups: males (over 4 h post-VEH: F(1,4) = 0.6077 and p = 0.4792; over 4 h post-CNO: F(1,4) = 16.40 and p = 0.0155) and females (over 4 h post-VEH: F(1,21) = 5.246 and p = 0.0238; over 4 h post-CNO: F(1,21) = 25.11 and p < 0.0001). C, I, RER over 24 h after sequential VEH and CNO injection in male or female mCherry or hM3Dq mice. RM two-way ANOVA between mCherry and hM3Dq groups: males post-CNO: F(1,8) = 22.04 and p = 0.0016; females post-CNO: F(1,21) = 5.594 and p = 0.0277. D, J, Locomotor activity average during 1-h period before and after VEH or CNO injection in male or female mCherry or hM3Dq mice. Paired t test: ns = not significant, *p < 0.05, and **p < 0.01. E, K, Oxygen consumption average during 1-h period before and after VEH or CNO injection in male or female mCherry or hM3Dq mice. Paired t test: ns = not significant, **p < 0.01, and ****p < 0.0001. F, L, RER average during hours 8–12 in male or female mCherry or hM3Dq mice. Unpaired t test: ****p < 0.0001. Data are expressed as mean ± SEM. Animal number in each group is listed in legends.