The aim of this study was to increase our understanding of the physiological functions controlled by the endocannabinoid system during embryogenesis. Using genomic and proteomic methodologies applied to zebrafish, we proved, for the first time in an oviparous species, that the cannabinoid receptor CB1 is not a maternal factor. The analysis of different developmental stages showed that the zygotic expression of CB1 occurs from the 3 somites stage while CB1 protein becomes evident during hatching time, indicating an involvement in the hatching process. This result was supported by the data regarding embryo exposure to the CB1 antagonist, AM251, consisting in a 75% decrease in hatching rate. In addition, as previously described for mammals, we observed a role of CB1 in the motility behavior in zebrafish larvae.