Immunohistochemical localization of neurocan in the lower auditory nuclei of the dog

Chondroitin sulfate proteoglycans are present at high levels in the lower auditory system of mammals. Axon terminals on the principal neurons in the superior olivary nuclei contain chondroitin 4- and 6-sulfate, while the broad extracellular matrix around axon terminals contains chondroitin sulfate D, a highly sulfated chondroitin sulfate rich in the disaccharide unit of GlcA(2S)beta1 --> 3GalNAc(6S), in the dog. In the present study, we investigated the immunohistochemical staining of neurocan, a brain-specific proteoglycan, in the lower auditory tract of the dog, including an analysis by immunoelectron microscopy. Immunolocalization of neurocan was conspicuous in the medial and lateral superior olivary nuclei and much less intense immunostaining was seen in the cochlear nucleus and posterior colliculus. No immunoreactivity were found in other nuclei. The immunostaining in the medial and lateral superior olivary nuclei was observed as perineuronal nets around large principal neurons at the light-microscopic level, while no immunostaining was observed in the upper segment of the medial superior olivary nucleus and the medial segment of the lateral superior olivary nucleus, in which medium-sized and small neurons were located. Immunoelectron microscopy revealed the reaction products of immunostaining on cell membranes of the perikarya of principal neurons and on cell membranes of presynaptic terminals which made axo-somatic synapses on the principal cells. No immunoreactivity was detected at synaptic junctions, in the extracellular matrix or within axon terminals. In the cochlear nucleus, immunoreactive perineuronal nets were found around a small number of neurons and immunoreactive nerve fibers were scattered in the anterior ventral cochlear nucleus. In the posterior colliculus, perineuronal nets, which were weakly immunostained, were sparsely distributed in the central nucleus. These results suggest that different locations of chondroitin sulfate proteoglycans, including neurocan, may be associated with focal sites composed of neuronal surface, terminal boutons and extracellular matrix in the lower auditory tract of the adult dog.