The olfactory epithelium, which retains a capacity for neurogenesis throughout life, contains two categories of basal cells, dark/horizontal and light/globose, neither of which is fully characterized with respect to their function during the processes of neurogenesis and epithelial reconstitution after injury. The aim of this study was to define the potential biological role(s) of dark/horizontal basal cells (D/HBCs) in the epithelium by performing immunochemical, electron microscopic, and developmental analyses of this cell population. The D/HBCs express several specific immunochemical characteristics, which include the rat homologues of human cytokeratins 5 and 14, which were identified on the basis of staining with subunit-specific monoclonal antibodies and two-dimensional immunoblot analysis of the immunoreactive proteins. Indeed, the D/HBCs are the only cells in the olfactory mucosa that express these specific cytokeratins. The D/HBCs also express an alpha-galactose or alpha-N-acetyl galactosamine moiety to which the I beta 4 isolectin from Bandeiraea simplicifolia binds. Moreover, the D/HBCs are heavily labeled by two different antibodies against the EGF receptor and by a monoclonal antibody that binds to phosphotyrosine. These characteristics are also common to the basal cells of respiratory epithelium. The electron microscopic analysis of the basal region of the olfactory epithelium and the light microscopic immunofluorescence observations demonstrate that the D/HBCs provide a bridge between the basal processes of some sustentacular cells and the basal lamina. The most striking ultrastructural feature of the D/HBCs is their enfolding of virtually all bundles of olfactory axons within tunnels formed where D/HBCs arch over the basal lamina. The intimacy of the arrangement between D/HBCs and olfactory axons suggests that signals may pass from axons to D/HBCs or vice-versa. With respect to the development of D/HBCs, cells that express cytokeratins 5 and 14 and the EGF receptor first appear near the boundary with respiratory epithelium late in development, but do not extend throughout the olfactory epithelium until the middle of the first postnatal week. Taken together, the present findings and previously published data suggest that D/HBCs help to maintain the structural integrity of the olfactory epithelium, participate in its recovery from injury, and may also function to signal the status of the neuronal population of the epithelium.