Figure 1.
Distinct neuronal populations in the BNST recapitulate IL-1β responses. (A) STPT of the whole mouse brain of TRAP2-tdTomato mice. Top lane: overview of the STPT data acquisition and registration; middle lane: representative coronal sections; and bottom lane: areas showing tdTomato+ neuronal cell bodies. (B) tdTomato+ cell count in indicated brain region in IL-1β (5 μg/kg; n = 6) and PBS-injected groups (n = 6). PVT, paraventricular nucleus of the thalamus; PBNc, parabronchial nucleus complex; LC, locus coeruleus; AP, area postrema; NTS: nucleus of the tractus solitarius. Data are represented as individual mouse data points with mean ± SEM. This experiment was completed once. One-way ANOVA with Tukey’s multiple comparison test. (C) Representative images showing immunofluorescence staining of the labeled cells (tdTomato, red) co-stained with DAPI (blue) in the BNST. tdTomato+ cell count in the dorsal and ventral BNST regions after exposure to IL-1β. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. Scale bar, 100 μm. (D) Schematic of TRAPing IL-1β–responsive BNST neurons and their reactivation in TRAP2 mice. (E) Representative images of BNST showing Gq-DREADD-mCherry–expressing cells (red) co-stained with c-Fos (green) after CNO administration. Scale bar, 20 μm. (F) Core body temperature for 6 h after reactivation with saline (black) as a control or CNO (red). (G) Minimum change in core body temperature at 1 h after injection. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (H) ΔHR for 60 min after reactivation of IL-1β–TRAPed neurons with saline (black) or CNO (red). Saline, n = 8 mice; CNO, n = 10 mice, mixed-effects analysis with Šidák correction. (I) AUC of ΔHR after reactivation. Data are represented as individual mouse data points from three independent experiments. Unpaired t test. (J and K) Serum levels of IL-6 and corticosterone at 2 h after saline or CNO administration. Data are represented as individual mouse data points pooled from three independent experiments. Unpaired t test. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Box and whisker plots show the minimum, maximum, median, and 25th and 75th percentiles.

Distinct neuronal populations in the BNST recapitulate IL-1β responses. (A) STPT of the whole mouse brain of TRAP2-tdTomato mice. Top lane: overview of the STPT data acquisition and registration; middle lane: representative coronal sections; and bottom lane: areas showing tdTomato+ neuronal cell bodies. (B) tdTomato+ cell count in indicated brain region in IL-1β (5 μg/kg; n = 6) and PBS-injected groups (n = 6). PVT, paraventricular nucleus of the thalamus; PBNc, parabronchial nucleus complex; LC, locus coeruleus; AP, area postrema; NTS: nucleus of the tractus solitarius. Data are represented as individual mouse data points with mean ± SEM. This experiment was completed once. One-way ANOVA with Tukey’s multiple comparison test. (C) Representative images showing immunofluorescence staining of the labeled cells (tdTomato, red) co-stained with DAPI (blue) in the BNST. tdTomato+ cell count in the dorsal and ventral BNST regions after exposure to IL-1β. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. Scale bar, 100 μm. (D) Schematic of TRAPing IL-1β–responsive BNST neurons and their reactivation in TRAP2 mice. (E) Representative images of BNST showing Gq-DREADD-mCherry–expressing cells (red) co-stained with c-Fos (green) after CNO administration. Scale bar, 20 μm. (F) Core body temperature for 6 h after reactivation with saline (black) as a control or CNO (red). (G) Minimum change in core body temperature at 1 h after injection. Data are represented as individual mouse data points pooled from two independent experiments. Unpaired t test. (H) ΔHR for 60 min after reactivation of IL-1β–TRAPed neurons with saline (black) or CNO (red). Saline, n = 8 mice; CNO, n = 10 mice, mixed-effects analysis with Šidák correction. (I) AUC of ΔHR after reactivation. Data are represented as individual mouse data points from three independent experiments. Unpaired t test. (J and K) Serum levels of IL-6 and corticosterone at 2 h after saline or CNO administration. Data are represented as individual mouse data points pooled from three independent experiments. Unpaired t test. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Box and whisker plots show the minimum, maximum, median, and 25th and 75th percentiles.

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