It is well known that fish embryos are vulnerable to waterborne nanoparticles (NPs), with delayed hatching being the most common and sensitive endpoint. Up-regulation of hatching enzymes has been believed to be an important detoxification mechanism for NPs, but the inner mechanism for such phenomena has been seldom investigated. This study aimed to investigate the role of pregnane X receptor (Pxr) in maintaining the robustness of embryo hatching after treatment with gold nanoparticles (AuNPs, 4 and 82 nm). For this purpose, embryos from mating of 6-month-old wild-type (WT) AB strain zebrafish (Danio rerio, 3∼4-cm-length) were treated with AuNPs since 4 h post-fertilization (hpf). It was found that both AuNPs significantly inhibited embryo hatching after 52-h treatment, with Au-4 being more toxic at the same mass concentrations. At non-toxic concentrations and median effective concentrations (EC