Insulin-like growth factor-binding proteins (IGFBPs) play important roles in downregulating IGF activity and growth and development in vertebrates under hypoxic stress. However, the mechanisms of hypoxia regulation of IGFBPs in teleost fishes are unknown. The involvement of reactive oxygen species (ROS) and hypoxia-inducible factors (HIFs) in hypoxia upregulation of IGFBPs in Atlantic croaker were investigated. Three croaker IGFBPs, IGFBP-1, IGFBP-2, and IGFBP-5, were cloned and characterized. Chronic hypoxia exposure [dissolved oxygen (DO): 1.7 mg/l for 24 wk] caused significant increases in hepatic and neural IGFBP-1 mRNA expression compared with tissue mRNA levels in fish held under normoxic conditions (6.5 mg DO/l). Moreover, longer-term chronic hypoxia exposure (22.7 mg DO/l for 1520 wk) caused significant increases in mRNA levels of all three IGFBPs in both liver and brain tissues. Hypoxia exposure also markedly increased superoxide radical (O2·?, an index of ROS) production and HIF-1? mRNA and HIF-2? protein expression in croaker livers. Pharmacological treatment with an antioxidant attenuated the hypoxia-induced increases in O2·? production and HIF? mRNA and protein expression as well as the elevation of IGFBP-1 mRNA levels. These results suggest that the upregulation of IGFBP expression under hypoxia stress is due, in part, to alterations in the antioxidant status, which may involve ROS and HIFs.