BOTH SURVIVAL AND DEVELOPMENT OF SPONTANEOUSLY ACTIVE-RAT HYPOTHALAMIC NEURONS IN DISSOCIATED CULTURE ARE DEPENDENT ON MEMBRANE DEPOLARIZATION

Suppression of endogenous electrical activity was found to have an adverse effect on the survival and bioelectric development of dissociated, embryonic rat hypothalamic neurons in long-term culture. Cultures were treated during the first two weeks in vitro with tetrodotoxin (TTX), a selective blocker of voltage-gated sodium channels, alone and in combination with high extracellular KCl ([K+]o), a membrane depolarizer. Neuron survival was assessed through cell counting experiments, while the development of spontaneous electrical activity was examined with extracellular, patch-electrode recordings. TTX caused both a decrease in cell survival and a decrease in spontaneously active cells; concurrent treatment with K+ protected cells from the adverse effects of TTX. K+ treatment alone increased the fraction of spontaneously active neurons without significantly affecting cell survival. When taken together, these results suggest that the long-term survival of active cells depends on continual membrane depolarization. From these observations, we conclude that there exists two populations of neurons: the electrically active population, whose survival is sensitive to electrical activity, and the quiescent population, whose survival is not.