Electrophysiological mapping was used to study frequency representation in the inferior colliculus (IC) of the mouse. In the lateral nucleus (LN) only part of the frequency range of hearing was represented and tonotopicity was separate from that in the rest of the IC. Highest frequencies occupied the medial part (M) of the central nucleus (CN). A single complete representation of the hearing range was present only if representations in the dorsal cortex (plus dorsomedial nucleus) and CN (including M) were combined. Continuous isofrequency planes making up these nuclei (without the lateral part of the CN) were reconstructed. They tilted from medial to lateral and from caudal to rostral. The steepness of the slopes increased from caudal to rostral and from dorsal to ventral (i.e., with increasing frequency). Isofrequency planes had similar angles of deviation from the horizontal plane as described for dendritic laminae in the CN. Differences of mapping in the lateral part of the CN from that in the rest of the CN could be explained by the different organization of laminae in this part. The relative amounts of IC depth and volume occupied by parts of the mouse audible frequency range were quantified. Frequency representation along IC depth was not proportional to that along cochlear length. Compared with the relative density of afferent nerve fiber supply within given frequency ranges represented along the basilar membrane, there is a relative under-representation in the IC up to 15-20 kHz and an over-representation of higher frequencies. Highest absolute tone sensitivity (lowest threshold) was found in neurons forming a column (running perpendicular to isofrequency planes) in the center of the IC. Results are discussed with regard to frequency representation, intrinsic neuronal organization, and functional segregation in the IC of mammals.