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The cochlear microphonics (CM), endocochlear potential (EP) and potassium ion activities in the endolymph and perilymph were measured in guinea pigs which received daily successive intramuscular injections of 500 mg/kg of body weight of kanamycin sulfate. Preyer’s reflex threshold at 8 kHz began to increase after the 5th day of kanamycin treatment and disappeared on the 11 th day. The maximum output of CM at 8 kHz began to decrease with a time course similar to Preyer’s reflex. The potassium ion activities in the endolymph and perilymph and the EP did not change appreciably through all experimental days. The magnitude of the negative EP decreased in parallel with the reduction in CM and the relative potassium conductance (G_k) between the endolymph and perilymph. These results lend support to the hypothesis that the site of production of the negative EP is probably in the hair cells and that the negative EP is mainly dependent on the permeability of the potassium ions in the organ of Corti.