Fig. 1: Stimulation of BNST projections to LHA evokes energy state-dependent reward seeking.

A Experimental schematic. AAV-syn-ChR2-eYFP was injected in BNST and optical fibers were implanted bilaterally above the LHA in WT mice. B Histological validation depicting ChR2-eYFP expression in BNST and optic fiber placement above the LHA. ac anterior commissure, EP entopeduncular nucleus, f fornix, LV lateral ventricle. C Example whole-cell current clamp recording from a ChR2-expressing BNST neuron. Light pulses (blue) reliably evoked action potentials. D Probability of spiking in response to pulse trains of blue light (n = 12 neurons). E Schematic of experimental design for optogenetic stimulation in head-fixed mice. F Raster plots of licking during fed and fasted conditions across a range of stimulation frequencies. Stimulation was delivered during blue shaded region. G Licking during the stimulation period increased with fasting for the ChR2 group (two-way ANOVA: main effect of Satiety: F(1,11) = 9.08, p = 0.01; main effect of Frequency: F(3,33) = 8.54, p = 0.0002; Interaction: F(3,33) = 4.44, p = 0.01). H Latency to lick from onset of stimulation for ChR2 group (two-way ANOVA: main effect of Satiety: F(1,11) = 18.74, p = 0.001; main effect of Frequency: F(3,33) = 26.94, p = 5.36 × 10⁻⁹; Interaction: F(3,33) = 6.072, p = 0.002). I Stimulation had no effect on licking in eYFP controls (two-way ANOVA: main effect of Satiety: F(1,7) = 15.45, p = 0.006; no main effect of Frequency: F(3,21) = 1.46, p = 0.26; no Interaction: F(3,21) = 0.16, p = 0.92). J Stimulation had no effect on latency to lick in eYFP controls (two-way ANOVA: main effect of satiety: F(1,7) = 56.93, p = 0.0001; no main effect of Frequency: F(3,21) = 0.19, p = 0.90; no Interaction: F(3,21) = 0.67, p = 0.58). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 Sidak’s multiple comparison’s post hoc test. n = 12 ChR2 and 8 eYFP mice. Data represent mean ± SEM. Source data are provided as a Source Data file.