Extended Data Fig. 7: Synthetic DRD2 agonist protects against a mouse model of EHEC infection using C. rodentium, strain DBS100, whereas a DRD2 antagonist blocks the effects.
From: Dopamine receptor D2 confers colonization resistance via microbial metabolites
a–i, Drd2fl/fl x Villin (Vil)-Cre or Drd2fl/fl mice were pre-treated with DRD2 agonist sumanirole (SUM, 4 mg/kg, IP injection) daily for 2 d. j–r, Drd2fl/fl x Vil-Cre or Drd2fl/fl mice were pre-treated with DRD2 antagonist L-741,626 (1 mg/kg, IP injection) daily for 2 d, followed by conventional (2 g Trp/kg diet, ad libitum) or Trp (42 g Trp/kg diet, ad libitum) diet for 7 d. a–r, The mice were then administered C. rodentium (CR, oral gavage, 108 colony-forming units, CFU) with continued (a–i) SUM treatment or (j–r) L-741,626 and Trp feeding. a, j Timeline for (a) SUM and (j) L-741,626 study. b–c, k–l, Bacterial load in (b, k) feces and (c, l) colon tissue was measured (b, k) every 1–2 d for 10 d post-infection and (c, l) at the peak of infection, 10 d post-infection. (d–f, m–o) Colon sections were stained with H&E and (d, m) blindly scored for submucosal edema (0-3), goblet cell depletion (0-3), epithelial hyperplasia (0-3), epithelial integrity (0-4), and neutrophil and mononuclear cell infiltration (0-3). Data are expressed as the sum of these individual scores (0-16). See Methods for full description of scoring rubric. (e, n) Crypt heights were measured. (f, o) Representative images. Scale bar: 50 μm. (g, p) Intestinal cryosections were stained with DAPI and Alexa Fluor 647-phalloidin. Pedestal formation = # of pedestals per host cell (SUM, n = 883; L-741,626, n = 1775 cells examined over 3 independent experiments). For box plots, interquartile ranges (IQRs, boxes), median values (line within box), whiskers (lowest and highest values within 1.5 times IQR from the first and third quartiles), and outliers beyond whiskers (dots), are shown. h–i, q–r, Intestinal epithelial cells were isolated, and cell lysates were analyzed by Western blotting with the indicated antibodies. Source data are provided in Supplementary Fig. 9. (q) Samples derive from the same experiment, and Western blots were processed in parallel. (i, r) Densitometry was performed using FIJI, n = 3 biological replicates. Data are representative of at least 3 independent experiments, n = 10 mice per group, bars = mean, error bars = standard deviation. Statistical analysis was performed using the two-tailed Student’s t-test (b, k) or one-way ANOVA, followed by post-hoc Tukey multiple comparison test: ***p < 0.001.
