Fig. 1
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Effect of the endocannabinoid receptor antagonists AM251 and AM630 on zebrafish embryo morphology. (A) Schematic showing the timeline for drug exposures and when experimental measurements were made. (A) Drug exposure during the first 24?h of development (0?24?hpf) and drug exposure during the second 24?h of development (24?48?hpf) are highlighted by the yellow bar. Embryos were allowed to develop in normal egg water after each treatment. Primary or secondary motor neuron axonal branching was investigated between 48 and 52?hpf, while locomotion was recorded at 120?hpf (5?dpf). Gross morphological observations occurred at 2 and 5 dpf. The treatments include vehicle control (0.1% DMSO), AM251 0.5?µmol l?1, AM630 5?µmol l?1 or AM251 0.5?µmol l?1+AM630 5?µmol l?1, either from 0 to 24?hpf or 24 to 48?hpf. (B,C) Representative images of treated embryos were taken at 48?52?hpf; scale bars: 0.5?mm. (D,E) Body length (mm) at 2 dpf development (n=33, 31, 40 and 35 for vehicle control, 0.5?µmol l?1AM251, 5?µmol l?1 AM630 and 0.5?µmol l?1 AM251+5?µmol l?1 AM630 treatments, respectively, from 0 to 24?hpf, and n=29, 27, 37 and 34 for vehicle control, 0.5?µmol l?1 AM251, 5?µmol l?1AM630 and 0.5?µmol l?1 AM251+5?µmol l?1 AM630 treatments, respectively, from 24 to 48?hpf). Data are presented as means±s.e.m. aSignificantly different from vehicle control, P<0.05; bsignificantly different from 0.5?µmol l?1 AM251, P<0.05; csignificantly different from 5?µmol l?1AM630, P<0.05 (one-way ANOVA followed by Tukey?s post hoc multiple comparisons test).