Mechanosensory hair cells of the inner ear are susceptible to death when exposed to a variety of drugs including aminoglycoside antibiotics. During avian and mammalian development, there is a period of relative insensitivity to aminoglycoside-induced hair cell death. This study was designed to test the hypothesis that zebrafish (Danio rerio) have developmental differences in sensitivity to aminoglycoside-induced hair cell death in the lateral line neuromasts. Larval zebrafish of various ages were exposed to several concentrations of neomycin, and their hair cells were examined using the potentiometric vital dye, DASPEI. Results indicate that zebrafish larvae aged 4 days post-fertilization are relatively insensitive to aminoglycoside-induced hair cell death compared to older fish. Thus zebrafish hair cells show developmental differences in sensitivity to aminoglycoside-induced death similar to those reported for inner ear hair cells of birds and mammals.