The preoptic area contains thermosensitive neurons, thought to be important in thermoregulation, and steroid-sensitive neurons, thought to be involved in reproduction. The preoptic area also contains osmosensitive neurons, considered important in water balance, and glucosensitive neurons, thought to function in the regulation of glucose. If these various neurons belong to separate populations, one might predict that most osmosensitive, glucosensitive, and steroid-sensitive neurons constitute the population of temperature-insensitive neurons rather than thermosensitive neurons. To test this hypothesis, single unit activity was recorded in preoptic tissue slices prepared from male rats. In addition to temperature changes, neuronal responses were examined with various perfusion media containing testosterone or estradiol (30 pg/mL), low glucose (1.0 mM), and increased osmotic pressure (309 mosmol/kg). It was found that the steroid-sensitive, osmosensitive, and glucosensitive neurons were not confined to the temperature-insensitive neurons; but that nearly half of the thermosensitive neurons responded to these nonthermal stimuli. This lack of specificity was also observed between osmosensitive and glucosensitive neurons; however, most of the steroid-sensitive neurons were highly specific for either estradiol or testosterone. Although these findings do not suggest a strong functional specificity for preoptic neurons, they do support studies emphasizing interactions between regulatory systems.