Fig. 4.
Adrenergic activation of thermogenic fat regulates colonocyte gene expression and intestinal disease tolerance. (A and B) Thermoneutral C57BL/6J female mice were treated with β3-adrenoreceptor agonist CL 316,243 or vehicle for 3 wk. Rate of oxygen consumption (VO2; A) and total activity (B) in female mice housed at Ta = 30 °C and treated with vehicle or CL 316,243 (n = 4 to 5 per treatment). (C) Heat map, GO enrichment analysis, and corresponding P values of differentially expressed genes in BAT of CL 316,243 and DSS-treated C57BL/6J male mice housed at Ta = 22 °C or Ta = 30 °C (n = 4 per temperature and condition, fold change ≥1.5, adjusted P value <0.05). (D and E) C57BL/6J male mice were treated with β3-adrenoreceptor agonist CL 316,243 for 2 wk at Ta = 30 °C prior to administration of DSS. Body mass (D) and colon shortening (E) in treated mice [n = 4 to 8 per condition; data analyzed by two-way ANOVA (D) and Student’s t test (E)]. (F) PCA of colonocyte transcriptomes from male C57BL6/J mice housed at Ta = 22 °C or Ta = 30 °C and treated with CL 316,243. Each dot indicates an individual biological replicate (n = 4 per temperature and condition). (G) Heat map, GO enrichment analysis, and corresponding P values of differentially expressed genes in colonocytes of C57BL/6J male mice treated with vehicle or CL 316,243 and administered DSS at Ta = 22 °C and Ta = 30 °C (n = 4 per temperature and condition, fold change ≥1.5, adjusted P value <0.05). (H–J) 16S rRNA-sequencing was performed on fecal samples from C57BL/6J male and female mice that were treated with CL 316,243 or vehicle for 3 wk (n = 9 per time point and temperature). Beta diversity was measured by weighted UniFrac (H; analyzed by ADONIS, Ta = 22 °C vs. Ta = 30 °C vehicle, R2 = 0.2227, P = 0.0016; 30 °C vehicle vs. 30 °C CL 316,243, R2 = 0.1952, P = 0.0096; 22 °C vs. 30 °C CL 316,243, R2 = 0.1914, P = 0.0133), alpha diversity was measured by the Shannon index (I; analyzed by paired Student’s t-test), and a volcano plot depicts significantly altered sequencing variants after treatment with CL 316,243 at Ta = 30 °C (J; analyzed by ALDEx2 with Wilcoxon rank-sum test, log2 fold change >1 and FDR <0.1). (K and L) Representative images (K) and quantitation (L) of pimonidazole staining of the large intestine in C57BL6/J male mice treated with β3-adrenoreceptor agonist CL 316,243 or vehicle at Ta = 22 °C or Ta = 30 °C and DSS for 3 d (n = 5 to 7 per group, analyzed by Student’s t test). Data are presented as mean ± SEM.