In most mammals, the superior olive is the first stage for binaural interaction. Neurons in the medial superior olive (MSO) receive excitatory input from both ears and are sensitive to interaural time or phase differences of low-frequency sounds. The mustached bat (Pteronotusparnelli parneffii), a small echolocating species with high-frequency hearing, probably does not use interaural time or phase differences as cues for sound localization. Although the mustached bat has a large MSO, there is some evidence that it is functionally different from the MSO in nonecholocating mammals. Most MSO neurons in the mustached bat are monaural, excited by a contralateral sound. Their responses are phasic and correlated with either the onset or the offset of a sound. As a first step in determining the origin of these phasic monaural responses, we traced the connections of the MSO by using both retrograde and anterograde transport methods. Excitatory inputs to the MSO originate from spherical cells in the anteroventral cochlear nucleus, almost exclusively from the contralateral side. Glycinergic inhibitory input is relayed from the contralateral cochlear nucleus through the medial nucleus of the trapezoid body. To investigate the interactions of the contralateral excitatory and inhibitory inputs at the level of the MSO cell, we recorded sound-evoked responses and applied glycine or its antagonist by using microiontophoresis. The results show that the phasic response to a contralateral sound is created by interaction of a sustained excitatory input with a sustained inhibitory input, also from the contralateral ear. Whether the response is to the onset or offset of a sound is determined by the relative timing between the excitatory and inhibitory inputs. Thus, in MSO of the mustached bat, the ipsilateral excitatory pathway from the cochlear nucleus seen in animals with low-frequency hearing is virtually absent, and the MSO is adapted for timing analysis by using input from only the contralateral ear.The mammalian superior olivary complex is the first stage at which there is convergence of inputs from the two ears. The current view of its function is that it provides two parallel pathways to the auditory midbrain, each of which is specialized to compare sound at the two ears and transmit basic information necessary for sound localization. In this scheme, neurons in the lateral superior olive (LSO) are selective for interaural sound level differences and neurons in the medial superior olive (MSO) are selective for interaural time or phase differences (1, 2).This view of MSO function is based mainly on data obtained in mammals with large heads and good lowfrequency hearing. Species with small heads and highfrequency hearing probably do not have interaural time or phase differences in a range that would be useful for sound localization, and in some cases they appear to lack an MSO (1, 3, 4). However, in other cases, notably in echolocating bats, a large MSO is present despite a small head size and high-frequency hearing rang...