Differentiation of a cell line from olfactory epithelium is induced by dibutyryladenosine 3′, 5′-cyclic monophosphate and12-O-tetradecanoylphorbol-13-acetate

Abstract

Olfactory receptor neurons undergo continual replacement with new cells differentiating from a population of basal cells located in the olfactory epithelium. We have previously described the isolation of a cell line from murine olfactory epithelium (MOE CL1) which is now shown to be sensitive to two differentiation promotion agents, dibutyryladenosine 3′, 5′-cyclic monophosphate (db-cAMP) and12-O-tetradecanoylphorbol-13-acetate(TPA). Cells grown in the presence of db-cAMP or TPA, after a lag phase of 72–96 h, became bipolar, with long neurite-like processes. Cells conditioned for at least one passage in db-cAMP or TPA, or cells exposed to a mixture of db-cAMP and TPA, attained a bipolar morphology within 24 h. Cultures grown with db-cAMP, TPA or db-cAMP + TPA showed significant increases in the percentage of process-bearing cells. An increase in population doubling time and decrease in saturation density were also observed in cultures exposed to db-cAMP. Cells exposed to TPA continued to divide at the same rate as control cells, while those treated with db-cAMP + TPA underwent terminal differentiation. Peripheral-type benzodiazepine binding sites, which within the olfactory epithelium are localized on the receptor neurons, were present in homogenate membranes of undifferentiated cells. Cells differentiated by exposure to db-cAMP + TPA for 7 days showed a 65% increase in the density (B max) of peripheral-type benzodiazepine binding sites. Undifferentiated cells in control cultures showed no spontaneous changes in membrane potential or regenerative responses during current injection. Small, bipolar cells in morphologically differentiated cultures, however, were capable of generating several types of spontaneous changes in membrane potential including slow, regular oscillations; irregular, slow and fast depolarizations; and trains of action potentials.