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The aim of the present study was to investigate the role of ovarian steroids in the opioid regulation of LH and prolactin release in mares. Effects of the opioid antagonist naloxone on LH and prolactin secretion were determined in ovariectomized pony mares. The animals were pretreated with either progesterone (500 micrograms kg-1) or oestradiol benzoate (10 micrograms kg-1) for 8 days and subsequently with a combination of progesterone and oestradiol for an additional 8 days. Naloxone administration (0.5 mg kg-1 i.v.) resulted in a significant release of LH as well as prolactin in mares after pretreatment with either oestradiol benzoate or progesterone plus oestradiol benzoate (P < 0.05). No significant changes in LH and prolactin secretion were detected in progesterone-treated and non-steroid-treated ovariectomized mares. These results indicate that a prolonged oestrogen influence activates the opioid inhibition of LH and prolactin release in mares. In contrast to other species, progesterone alone does not activate a tonic opioid inhibition of LH and prolactin secretion, but modulates the effect of oestrogens. The opioid systems therefore seem to be regulated by a sequence of different steroid environments, as found during the oestrous cycle. The parallel increases in prolactin and LH secretion in mares may indicate a common regulatory pathway for these two hormones.