Fig. 7: NPY^DRN/vlPAG neurons regulate feeding behaviors and positive valence via different neural pathways.

AAV virus-based anterograde tracing. a Representative images showing mGFP and mRuby expression in NPY^DRN/vlPAG neurons. b Representative images showing NPY^DRN/vlPAG neuron-derived projections (red) in multiple brain regions. The experiment was repeated 4 times with similar results. c 1-h food intake in mice fed ad libitum before and during optogenetic activation of NPY^DRN/vlPAG neuronal terminals in downstream brain regions. For the lateral hypothalamic area (LH) and the dorsomedial hypothalamic nucleus (DMH) stimulation data, two-sided Wilcoxon matched-pairs signed rank test; For the others, two-sided paired Student’s t test. The medial/lateral preoptic area (MPA/LPO): n = 6 mice; The paraventricular thalamic nucleus (PVT): n = 6 mice; LH: n = 8 mice; The anterior hypothalamic area/lateroanterior hypothalamic nucleus (AH/LA): n = 7 mice; DMH: n = 10 mice. d Percentages of time in the stimulation side of RTPP with NPY^DRN/vlPAG-terminals optogenetic activation. Repeated measures one-way ANOVA with post-hoc Tukey’s multiple comparisons test. MPA/LPO: n = 5 mice; PVT: n = 6 mice; LH: n = 6 mice; AH/LA: n = 5 mice; DMH: n = 7 mice. e Virus injection strategy to retrogradely label PVT- and LH- projecting NPY^DRN/vlPAG neurons and representative images showing PVT- (green) and LH- (red) projecting NPY neuronal terminals. f Representative images showing NPY^DRN/vlPAG neurons retrogradely labeled from PVT (green) and LH (red). Arrow indicates double-labeled neuron. Bottom: Total numbers of PVT- (green), LH- (red) projecting and PVT/LH- dual projecting NPY^DRN/vlPAG neurons (n = 4 mice). Representative traces and quantification of light-evoked postsynaptic current amplitude of NPY^DRN/vlPAG-PVT (g) or NPY^DRN/vlPAG-LH (h) connectivity with application of DNQX or picretoxin (PTX). The blue lines indicate 5-ms light pulse. In (g), n = 5 cells, repeated measures one-way ANOVA with post-hoc Tukey’s multiple comparisons test; In (h), n = 8 cells, two-sided Friedman test with post-hoc Dunn’s multiple comparisons test. Data are shown as mean ± SEM. ‘n’ refers to mice number. Source data are provided as a Source Data file.