Identification of an olfactory receptor neuron subclass: cellular and molecular analysis during development

Development of olfactory receptor neuron populations was studied using the previously described monoclonal antibody (Mab) 2B8 which binds to cell surface glycoproteins of presumptive olfactory receptor neurons. In order to definitively demonstrate that the cells recognized were olfactory receptor neurons and to better characterize these cells during development, a well-established receptor cell marker, olfactory marker protein (OMP), was studied at the same time as the 2B8 antigens in double-label immunofluorescence analyses of olfactory structures in rats from Day 13 of gestation (E13) to the early postnatal period. Olfactory epithelium cryostat sections of E13 rats showed binding of the 2B8 Mab to bipolar cells in caudal regions of the nasal cavity. The 2B8 Mab also recognized a large number of cells in the vomeronasal organ (VNO) at this stage. No specific binding of anti-OMP was seen until E15. At this time approximately half of the 2B8 reactive cells also expressed OMP. By birth, greater than 90% of the 2B8 reactive cells expressed OMP. The percentage of total fluorescent labeled cells which are double labeled remained relatively constant at 23-33% as the total number of cells increased between E15 and 2 days postnatal. 2B8 immunoreactivity can be found in the olfactory nerve layer of the olfactory bulb and the presumptive accessory olfactory bulb at E15. In double-label experiments the 2B8 Mab did not bind to all anti-OMP-labeled glomeruli of postnatal to adult rats. In summary, the 2B8 Mab recognizes cells early during development and appears to recognize a subclass of olfactory receptor cells and their axon terminals. Developmental changes in the electrophoretic profile of the olfactory 2B8 antigens were also studied. In the olfactory epithelium a single band at Mr of 200,000 was seen at E19. After birth three bands at 220,000, 180,000 and 110,000 were observed but in adults only two bands of Mr 215,000 and 163,000 were detected. During olfactory bulb development the Mr of the two major 2B8 reactive bands did not change but remained the same as the two major bands seen in the adult olfactory epithelium. The olfactory bulb band at Mr of 215,000 showed a 3 to 4-fold increase and the band at 163,000 showed a 10-fold increase in specific activity from birth to adulthood.