This study investigates the functional organization of the superior olivary complex of the mustached bat with classical transmission electron microscopy and postembedding immunocytochemistry for gamma-aminobutyric acid (GABA) and glycine antisera in semithin serial sections. The ultrastructure and distribution of terminal types in the lateral superior olive (LSO) and the medial nucleus of the trapezoid body (MNTB) closely resemble that of other mammals; the organization within the medial superior olive (MSO) differs significantly. The differences concern the relative proportion of putatively inhibitory boutons, which appear as symmetrical synapses with flattened vesicles on MSO somata. In the bat, inhibitory boutons comprised 75-100% of perisomatic boutons, a value identical to that observed in the LSO. These terminals most likely arise from the MNTB. In other species, putatively inhibitory terminals form a much smaller proportion of perisomatic boutons in MSO. This difference suggests that in the bat MSO excitatory input to cell somata is considerably reduced and outweighed by inhibitory input. This suggestion is corroborated by immunocytochemical data. Glycine-immunoreactive puncta encrust somata of LSO and MSO cells to a similar degree and in rather homogeneous patterns throughout these nuclei. Putatively GABAergic terminals are located mainly on distal dendrites of MSO and LSO cells. Regional variations in the density of GABA-immunoreactive puncta in LSO suggest that different tonotopic zones are under differential modulatory influence. Both the LSO and MSO of the mustached bat contain significant amounts of putatively inhibitory projection cells. Coexistence of both antigens was commonly observed in subsets of cells. Quantitative analyses of labeling patterns and comparisons with other mammals suggest that the mix of neurotransmitters in projection cells of LSO and MSO is phylogenetically flexible, and thus the details of the functions of ascending pathways are species specific. In contrast to other mammals, the bat MSO forms parallel output pathways with excitatory and inhibitory components. Data are discussed in relation to specialized physiological response features.