The development of the rat inner ear starts about day 9 of a 22-day gestational period. The sensory and supporting cells are morphologically identifiable by day 18 of gestation; however, auditory function is not evident until day 9 postnatally. The organ of Corti attains adult maturity about day 16 postnatally. Morphologic and physiologic studies have shown that administration of sublethal doses of aminoglycosidic antibiotics, including kanamycin, damages the sensory and supporting cells of the inner ear in mammals. This results in the impairment of auditory function. This study was designed to determine the earliest stage in cochlear duct development at which kanamycin toxicity can be morphologically detected. A sublethal dose of kanamycin was administered to pregnant rats on days 10 through 20 of gestation and to neonates from days 1 to 8 or from days 8 to 16 postnatally. The inner ears were dissected and processed for light microscopic and ultrastructural observations. The cochlear ducts of fetuses exposed in utero to kanamycin and those of neonates exposed from days to 1 to 8 postnatally were unaffected. Kanamycin toxicity was evident in the cochlear ducts of neonates exposed from days 8 to 16 postnatally. The most severe damage occurred in the basal coil of the cochlea where in many instances sensory cells were totally absent. The sensory cells in the middle and apical coils exhibited abnormal morphology. These results suggest: (1) kanamycin does not have any permanent toxic effects on the rat cochlear duct during gestation and the first week of postnatal life. This provides a basis for further studies into the possible pharmacologic application of this finding in humans. (2) A correlation exists between the initiation of auditory function and the vulnerability of the organ of Corti to kanamycin toxicity.