Characterization of BNST cells. (A) snRNA-seq cataloging neuronal clusters in the BNST. A uniform manifold approximation and projection (UMAP) plot of transcriptomic data identified 17 neuronal clusters. This experiment was completed once. (B) Dot plots of average expression of Slc17a6 (Vglut2; glutamatergic neuronal marker), Slc32a1 (Vgat: GABAergic neuronal marker), corticotropin-releasing crh, sst, pdyn, and penk-expressing populations. Dot size: percent of cells in cluster and color: average expression levels. (C) ΔHR for 60 min after activation of CRH⁺, SST⁺, PDYN⁺, or PENK⁺ BNST neurons with saline (black) as a control or CNO (red) (crh-cre; saline, n = 5 mice, CNO, n = 5 mice, sst-cre; saline, n = 5 mice, CNO, n = 5 mice, pdyn-cre; saline, n = 5 mice, CNO, n = 5 mice, penk-cre; saline, n = 3 mice, CNO, n = 3 mice, mixed-effects analysis with Šidák correction). (D) Serum IL-6 levels at 2 h after activation with saline or CNO of CRH⁺, SST⁺, PDYN⁺, or PENK⁺ BNST neurons. (E) Serum corticosterone levels at 2 h after activation with saline or CNO of CRH⁺, SST⁺, PDYN⁺, or PENK⁺ BNST neurons. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. *P < 0.05, **P < 0.01.