Fig. 1: NAcSh Serpinb2+ neurons are activated in the refeeding process in vivo.
a, Spatial patterns of D1-MSN subtypes in NAcSh. Dotted lines circle the anterior commissure olfactory limb (aco) and NAc core. The dorsal–ventral and medial–lateral axes are indicated. b, Spatial expression patterns of Stard5, Tac2, Spon1 and Serpinb2, determined by integrating single-cell RNA sequencing and MERFISH data using iSpatial. These genes are able to specifically label distinct neuronal subtypes in the medial dorsal NAc shell. Expression level is color-coded. c, RNA in situ hybridization showing Spon1, Stard5 and Drd1 expression in the medial part of the NAcSh. Boxed regions in the left panels are enlarged and shown in the right panels. Scale bars, 500 μm (left), 20 μm (right); n = 3 biologically independent experiments. d, RNA in situ hybridization showing Tac2, Seprinb2 and Drd1 expression in the medial part of the NAcSh. Boxed regions in the left panels are enlarged and shown in the right panels. Scale bars, 500 μm (left), 20 μm (right); n = 3 biologically independent experiments. e, Schematic representation of the experimental design for the ad libitum, fasted and refed groups. f, Co-expression of Spon1, Stard5, Tac2 and Serpinb2 mRNA with cFos mRNA in NAc of the ad libitum, fasted and refed states. Representative images showing the colocalization of cFos (green), Spon1 (magenta), Stard5 (red), Tac2 (magenta) and Serpinb2 (red) expressing neurons. Scale bars, 50 μm; n = 3 biologically independent experiments. g, The average number of cFos+ neurons in the dorsal medial NAcSh in the ad libitum, fasted and refed states (n = 5 sections from three mice of each group; one-way ANOVA with Tukey’s multiple comparison, F2,12 = 27.82). h, The percentages of activated Spon1, Stard5, Tac2 or Serpinb2-expressing neurons in the total cFos+ neurons in dorsal medial NAcSh in the refed state (n = 5 sections from three mice of each group; one-way ANOVA with Tukey’s multiple comparisons, F3,8 = 15.50). i, The percentages of activated Spon1, Stard5, Tac2 or Serpinb2-expressing neurons in the total Spon1, Stard5, Tac2 or Serpinb2-expressing neurons at ad libitum, fasted and refed states (n = 5 sections from three mice of each group; one-way ANOVA with Tukey’s multiple comparisons, F11,24 = 81.44). All error bars represent mean ± s.e.m.
