Fig. 5: Overnutrition transiently impairs BNST-mediated recruitment of LHA^Vglut2 neurons.

A Experimental schematic for HFD overfeeding. Mice were maintained on ad libitum HFD for 3 weeks prior to testing. B Example imaging plane (top, standard deviation projection) of LHA^Vglut2 neurons recorded after overfeeding in fed (left) and overnight fasted (right) mice (25.4 ± 8.2 cells per mouse). Heat maps show average response of all cells from all mice aligned to stimulation onset (bottom). Shaded area represents masked frames corresponding to light artifacts. C Average responses (±SEM) of all recorded neurons aligned to stimulation offset. D Average response immediately after the cessation of stimulation (3 frames, 400 ms) for neurons that were recorded in both fed and fasted conditions. Two-way ANOVA: no main effect of Satiety: F(1,256) = 0.003, p = 0.95, main effect of Frequency: F(3,256) = 109.6, p < 1.0 × 10⁻¹⁵, Interaction: F(3,256) = 2.77, p = 0.04. E Experimental schematic for post-HFD imaging. Following HFD, mice were returned to standard chow for 3 weeks prior to testing. F Example imaging plane (top, standard deviation projection) of LHA^Vglut2 neurons recorded after overfeeding in fed (left) and overnight fasted (right) mice (21.0 ± 5.3 cells per mouse). Heat maps show average response of all cells from all mice aligned to stimulation onset (bottom). Shaded area represents masked frames corresponding to light artifacts. G Average responses (±SEM) of all recorded neurons aligned to stimulation offset. H Average response immediately after the cessation of stimulation (3 frames, 400 ms) for neurons that were recorded in both fed and fasted conditions. Two-way ANOVA: main effect of Satiety: F(1,260) = 53.28, p = 3.52 × 10⁻¹², main effect of Frequency: F(3,260) = 143.2, p < 1.0 × 10⁻¹⁵, Interaction: F(3,260) = 14.85, p = 5.97 × 10⁻⁹. Sidak multiple comparison test, **p = 0.002, ****p < 0.0001. n = 7 mice. Source data are provided as a Source Data file.