To test the hypothesis that progesterone (P) administered in a high dose and for a long duration would influence in vitro LHRH release by naloxone, silastic capsules containing P (50 mg/ml) or vehicle were implanted sc in ovariectomized, estradiol-primed (OVX+E) prepubertal rats. Following a 48 hr exposure to P, rats were sacrificed and mediobasal hypothalamic (MBH) fragments were obtained. In addition, in other OVX+E primed rats, a single injection of P (1 mg) was given sc 6 hr prior to decapitation. Naloxone or control medium were infused into superfusion chambers containing MBH fragments derived from these animals for 2 hr following a 1 hr control period. Infusion of naloxone (1 x 10(-4) M) markedly stimulated in vitro LHRH release from MBH from rats pre-exposed to P for 48 hr, whereas it was unable to do so in other groups examined. These data clearly indicate that the ability of naloxone (1 x 10(-4) M) to stimulate LHRH release is dependent on the duration of P administration.