Freeze-fracture methodology was used to study rat hypothalamic arcuate nucleus (AN) neuronal plasma membrane organization following in vitro perfusion of brain slices with 17-beta-estradiol (17 beta E2) or other test compounds. Physiological levels (10(-10) M) of 17 beta E2 caused an increase in neuronal membrane exo-endocytotic pits within 1 min of perfusion. The increased density of pits was dose related, sustained at a constant rate during 10 min of perfusion, reverted to control values after perfusion with estradiol-free medium for 1 h, and was accompanied by an increased uptake of horseradish peroxidase by the arcuate nucleus in brain slices. The 17 beta E2-induced increase in exo-endocytotic pit density was blocked by tamoxifen (10(-8) M). Cholesterol (10(-10) M), 17-alpha-estradiol (10(-6) M) or dihydrotestosterone (10(-6) M) had no effect on exo-endocytotic pit density. Testosterone had about 50% the potency of 17 beta E2 in increasing exo-endocytotic pit density. These results indicate that physiological levels of 17 beta E2 can have rapid effects upon arcuate nucleus neuronal membrane ultrastructure.