A basic principle of organization in auditory centers is the topographic-tonotopic order. Whether this applies to the dorsal nucleus of the lateral lemniscus (DNLL), however, is still debated. To clarify this problem, we have utilized the neuroanatomical tracers horseradish peroxidase (HRP) and biotinylated dextran (BD) injected into different regions of the central nucleus of the inferior colliculus (CNIC) in the rat. After large injections of HRP that included most of the CNIC, retrogradely labelled neurons were found all across the ipsi- and contralateral DNLL, showing that all parts of this nucleus innervate the CNIC bilaterally. More neurons were seen consistently on the side contralateral to the injection site. Labelled fibers, however, were abundant ipsilaterally, but scarce in the contralateral DNLL. Single, small injections of HRP or BD into the CNIC resulted in labelling in restricted areas of the ipsi- and contralateral DNLL. In coronal sections, the neurons and fibers labelled in the ipsilateral DNLL formed a well-defined, ring-shaped structure made of dendrites and axons oriented parallel to each other, which we termed "annular band." The observation of serial sections revealed that the annular band seen in any individual section represents a slice through a more or less complete three-dimensional, hollow, ovoid structure oriented rostrocaudally. The position and diameter of the annular band changed as the injection site was shifted along the tonotopic axis of the CNIC. Single injections placed in the ventromedial, high-frequency region of the CNIC produced a large annular band along the periphery of the DNLL. After injections placed in progressively more dorsolateral, lower-frequency regions of the CNIC, the annular band became smaller in diameter and occupied a successively more central position in the DNLL. Double injections along the tonotopic axis of the CNIC resulted in two roughly concentric annular bands. The labelled neurons and fibers in the contralateral DNLL systematically occupied a position symmetric to the annular band seen ipsilaterally. These findings indicate that the rat DNLL is primarily composed of neurons with flattened dendritic arbors and flattened fields of terminal fibers. These two elements intermingle, forming concentric layers around the geometric center of the nucleus. The axons of neurons within corresponding layers on the two sides converge onto the CNIC of both sides in a strict topographic fashion: the peripheral layers project to the ventromedial, high-frequency region of the CNIC, and the central layers project to the dorsolateral, low-frequency region. These results suggest that the concentric arrangement of the DNLL is the substrate of its tonotopic organization.