We studied the sensitivity to a depolarizing stimulus of hypothalamic fragments dissected from cycling female donor rats exposed or not to 30-min stress at 4 degrees C. The neuronal response was estimated in terms of the ability of tissue to release somatostatin when stimulated with 40 mM K+. The data showed no differences in response to K+, regardless of the ovarian cycle of the female donors, whereas tissues dissected from ovariectomized or pregnant rats responded significantly to K+. However, when donors underwent previous cold stress, significant differences were noted at all stages of the cycle, except diestrus-1, compared with control rats. We tested whether GABA and/or neuroactive steroids could be involved in this phenomenon and observed no GABA inhibition of somatostatin release in vitro, but inhibition occurred in the presence of a neuroactive steroid, THDOC. The effect of GABA in vivo on somatostatin release was estrogen dependent because bicuculline modified the total amount of somatostatin secreted in estrus but not in diestrus II. Finally, in hypothalamic primary cultures, GABA inhibition of somatostatin release was only detected when steroids were present in the media throughout culture. Our results suggest that steroid-GABA-somatostatin interactions could explain the different responses of neurons to depolarization.