Paraffin embedded sections of rat, mouse, dog, and human brain were stained with a battery of lectin-horseradish perioxidase conjugates to localize and characterize glycoconjugates. In the rat and mouse cerebral cortex, a subpopulation of nonpyramidal neurons stained selectively with three lectins with specific affinity for terminal N-acetylgalactosamine (GalNAc). These and only these lectins stained the surface of the cell body, dendritic shafts, and proximal parts of the dendritic arborization. Most reactive, nonpyramidal neurons revealed a multipolar dendritic pattern, but some possibly belonged to the bitufted and bipolar types of neuron. The GalNAc-containing neurons appeared widely distributed in layers II-VI with relatively greater abundance in layers IV and V. In the cortex of rats and mice the stained neurons occurred in moderate numbers in the frontal, frontoparietal, striate, retrosplenial, and entorhinal regions, but were less numerous in the hippocampal gyrus, dentate gyrus, and olfactory area. Other neurons in the basal ganglia and brain stem stained weakly for GalNAc. Examination of the frontal cortex of human and canine brains showed a similar distribution of nonpyramidal neurons with affinity for GalNAc-binding lectins. At high magnification, the surface staining of neurons in the cerebral cortex, deep cerebellar nucleus, and other sites appeared periodic rather than continuous. The periodic character of the neuronal surface staining suggested a location for the reactive glycoconjugate in or between the synapses. The GalNAc-containing glycoconjugate occurred in a selected cell type, failed to bind the other lectin conjugates, and differed from biochemically detected glycoconjugates. It is, therefore, considered a newly recognized entity of possible physiologic significance for a population of cortical neurons.