We have described the acoustic pathway from the ear to the diencephalon in a sound-producing fish (Pollimyrus) based on simultaneous neurophysiological recordings from single neurons and injections of biotin pathway tracers at the recording sites. Fundamental transformations of auditory information from highly phase-locked and entrained responses in primary eighth nerve afferents and first-order medullary neurons to more weakly phase-locked responses in the auditory midbrain were revealed by physiological recordings. Anatomical pathway tracing uncovered a bilateral array of both first- and second-order medullary nuclei and a perilemniscal nucleus. Interconnections within the medullary auditory areas were extensive. Medullary nuclei projected to the auditory midbrain by means of the lateral lemniscus. Midbrain auditory areas projected to both ipsilateral and contralateral optic tecta and to an array of three nuclei in the auditory thalamus. The significance of these findings to the elucidation of mechanisms for the analysis of communication sounds and spatial hearing in this vertebrate animal is discussed.