Antimitotic drugs that enhance neuronal survival in olfactory bulb cell cultures

Cell cultures of olfactory bulb from 2-3-day-old rats were used to evaluate 6 antimitotic drugs for their effects on neuronal survival. In cultures grown in medium without drugs for 7 days, neurons died as a layer of non-neuronal cells proliferated. A series of experiments were performed to determine dilutions and exposure times that allow maximum neuronal survival for each of the antimitotic drugs. Use of bromodeoxyuridine (BUdR) at 10(-5) M with a 5 day exposure gave cultures with large aggregates of neurons and an extensive network of interconnecting neurites. Cytosine arabinoside (Ara-C) at 8 X 10(-6) M for 5 days of exposure was also effective but the aggregates contained cellular debris. Fluorodeoxyuridine (FUdR) at 10(-4) for 5 days was less effective in flowing neurons to survive. Methotrexate (MTX), high thymidine and hydroxyurea were all not effective in allowing neuronal survival. In all cases there was an inverse relationship between the survival of neurons and proliferation of non-neuronal cells. Additional experiments were performed to determine how proliferating non-neuronal cells can lead to the death of neurons. In a series of transfer experiments, cultures were exposed to conditioned medium. Cultured olfactory bulb neurons grown on small cover slips were exposed to BUdR under optimal conditions and after 7 days were transferred to cultures treated or not treated with BUdR. In cultures not treated with BUdR, most of the transferred BUdR treated neurons died, while in cultures treated with BUdR the BUdR treated neurons survived. These results suggest that antimitotics enhance olfactory bulb neuronal survival by reducing the number of non-neuronal cells. In addition, it appears that proliferating non-neuronal cells are responsible for neuronal cell death by a medium factor and not by contact with the dividing non-neuronal cells.