The localization of glycosylated molecules and glia has been studied during early postnatal development in the mouse central nervous system (CNS) using autoradiographic detection of radiolabeled fucose incorporation, and in sections processed either for histochemistry or immunocytochemistry following binding of labeled lectins or an antibody to glial fibrillary acidic protein. Radiolabeled sugar incorporation, lectin binding of glycoconjugates, and glial labeling all reveal borders between nuclei within the diencephalon, midbrain, and brainstem through the first postnatal week. Glycoconjugate and glial boundaries exist throughout the CNS during pattern formation events, and they also are seen in relation to fine aspects of developing functional organization within individual structures (e.g. segmentation associated with the representation of mystacial vibrissae within the brainstem trigeminal complex). The observation that each of the probes employed in this study fails to label boundary organization during later postnatal times suggests that the distribution and chemistry of the glial/glycoconjugate network are dynamic, and they change in accordance with distinct maturational states of the nervous system.