The ultrastructural distribution of gentamicin in the cochlea was investigated immunocytochemically. Specific labeling was restricted to the organ of Corti, in particular to the outer and inner hair cells, the Deiters' cells, Hensen's cells and the tympanic layer cells of the basilar membrane. Other cochlear tissues did not demonstrate any labeling. At the subcellular level, gentamicin was found in lysosomes, multivesicular bodies and small tubules and vesicles. A model is proposed in which it is hypothesized that gentamicin is internalized by endocytotic vesicles and is transferred to the lysosomal compartment as well as to the endoplasmic reticulum and Golgi complex.