Musk ambrette (4-tert-butyl-3-methoxy-2,6-dinitrotoluene) is a nitro musk, a cheap substitute for natural musk and a potential environmental pollutant based on its persistence, accumulation in human organisms. We investigated the acute toxicity of musk ambrette using wild-type AB and transgenic Tg(fli1a:EGFP)y1 zebrafish. Different concentrations were delivered to zebrafish by direct soaking from 6 to 72 h post-fertilization (hpf). The LC50 of musk ambrette was 76.4 µg mL−1. As musk ambrette concentration increased, zebrafish embryos showed developmental delays (50 µg mL−1, 22 hpf), pericardial edema (5 µg mL−1, 48 hpf), circulatory disturbances, curved body axis (1 µg mL−1, 72 hpf) and death (100 µg mL−1, 22 hpf). Target organ toxicity was evaluated by a zebrafish angiogenesis model. Musk ambrette induced cardiovascular morphological changes, vessel permeability variation, angiogenic changes and cardiotoxicity (10 µg mL−1, 48 hpf). The disappearance of caudal vein plexus confirmed the vascular development toxicity. Musk ambrette negatively affects early life-stage survival and demonstrates various toxicities in zebrafish.