Label localization was compared by light and electron microscopic autoradiography in the organ of Corti of the gerbil, following in vivo incubation with one of eight 3H-labeled amino acids. Following incubation with GABA, efferent endings underneath the outer hair cells were heavily labeled, as were all tunnel crossing fibers, many small fibers in the inner spiral bundle and some efferents beneath the inner hair cells. This pattern was absent following incubations with the GABA analog muscimol. Following L- and especially D-aspartic acid incubations, efferent endings under the inner hair cells and small fibers of the inner spiral bundle were preferentially labeled. It is concluded that these differential labeling patterns after GABA and aspartic acid incubations reflect two populations of efferent fibers in the organ of Corti: one terminating primarily upon the outer hair cells and one underneath the inner hair cells. Cochlear hair cells showed the highest level of labeling following incubation with alanine. Inner hair cells were more heavily labeled than outer hair cells following incubations with alanine and glycine. Hair cell labeling was comparable to that of supporting cells for most amino acids. However, for alanine and glycine the labeling of inner hair cells, and for L-aspartic and glutamic acid the labeling of all hair cells, was higher than that of structural cells. Labeling of hair cells was substantially lower than that of structural cells following taurine incubations.