Neuron-glia-related cell adhesion molecule (Nr-CAM) is a recently characterized cell adhesion molecule in the family of immunoglobulin-related molecules of which the neural cell adhesion molecule, N-CAM, is the prototype. Nr-CAM shares structural properties with another member of this family (neuron-glia CAM, Ng-CAM) and both molecules exhibit homophilic and heterophilic binding properties. To understand better the role of such molecules in development, we have examined the sites of synthesis and expression of Nr-CAM by means of in situ hybridization and immunohistochemistry. Both methods indicated that Nr-CAM is expressed only in the nervous system. The molecule was observed on neurons in both the peripheral and central nervous systems and on epithelial floor plate cells in the spinal cord, but it was absent in the germinal zones. The protein was present on perikarya, but was found preferentially on axonal tracts. As observed for messenger RNAs specifying other cell adhesion molecules, messenger RNA for Nr-CAM was localized in the perikarya. The temporal expression of Nr-CAM was correlated with various neural morphoregulatory events, including cell proliferation and migration, axonal outgrowth and myelination. The molecule was expressed during the onset of neurogenesis at embryonic day 3 in the floor plate epithelium, and then on postmitotic ventral horn motor neurons of the spinal cord. At later stages, it was expressed throughout the spinal cord but disappeared from the floor plate. In the cerebellum, Nr-CAM was found on granule and Purkinje neurons and afferent fibers. Both local and projection neurons in the optic tectum, as well as axonal pathways throughout the telencephalon, expressed Nr-CAM. In the peripheral nervous system, Nr-CAM was expressed strongly in sensory and autonomic ganglia and in the enteric nervous system. At the onset of myelination, there was a general decrease in staining for Nr-CAM protein in the central nervous system but not in the periphery. Comparison of the expression of Nr-CAM to that of the structurally related Ng-CAM showed considerable overlap in their distributions, although there were differences in the levels at which each CAM was observed in particular structures. For example, sympathetic ganglia stained more intensely for Nr-CAM protein than for Ng-CAM. This differential but co-distributed pattern is consistent with the idea that although similar cell adhesion molecules have independent binding specificities, they may have related functions that act synergistically in the development of the nervous system.