CB1 receptors mediate CB teratogenesis. (a) Effects of pretreatment with a CB1 receptor antagonist, SR 141716A, on CP 55,940-induced eye defects in mice. Mice received CB vehicle (n = 43/6 litters) SR 141716A 5.0 mg/kg + the CB vehicle (n = 62/8 litters), the CB vehicle + CP 55,940 0.5 mg/kg (n = 51/8 litters), SR 141716A 2.0 mg/kg + CP 55,940 0.5 mg/kg (n = 45/6 litters), or SR 141716A 5.0 mg/kg + CP 55,940 0.5 mg/kg (n = 88/10 litters). SR 141716A or CB vehicle was given 10 min before the second injection. **p < 0.01, ***p < 0.001 vs. CB vehicle, ^^p < 0.01 vs CP 0.5, using a Chi-square test. Data are the percent of mice with defects (number of affected mice/total number of mice *100). See Supplementary Table 4 for eye defect severity, body weight and length, and the number of live offspring/litter. See Supplementary Fig. 2a–f. for separate analysis of left and right eye defects. (b) Small eyes in zebrafish embryos receiving exposure to fish water vehicle (n = 39), SR 141716A 3.0 mg/L (n = 40), CP 55,940 5.0 mg/L alone (n = 54), SR 141716A 3.0 mg/L + CP 55,940 5.0 mg/L alone (n = 49), 0.5% alcohol + CP 55,940 2.5 mg/L (n = 42), SR 141716A 3.0 mg/L + 0.5% alcohol + CP 55,940 2.5 mg/L (n = 49). **p < 0.01, ***p < 0.001 vs. vehicle. ^^p < 0.01, ^^^p < 0.001 vs. corresponding treatment without SR 141716A using a Chi-square test. Data are the percent of zebrafish with defects (number of affected zebrafish/total number of zebrafish *100). (c,d) CB1-Smo co-localization in the mouse neural tube (GD 9.5) using a proximity ligation assay (Duolink) to visualize sites (fluorescent red) where anti-CB1 and anti-Smo antibodies bind within 40 nm of one another. (e,f). CB1-GPR 161 co-localization in the mouse neural tube (GD 9.5). Nuclei of neural tube cells are labeled with DAPI in blue. Sections (7 µm) were imaged on a confocal microscope using a 40× (c,e) and 63× (d,f) oil lens.
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