The effect of progesterone (P4) and P4 withdrawal on oestradiol (E2)-induced change in gonadotrope responsiveness (GR) and concentration of GnRH receptor and GnRH receptor mRNA in pituitary tissue of orchidectomized sheep (wethers) was determined. Thirty wethers were assigned at random to one of six treatment groups (n = 5 animals/group). Wethers received E2 (2 micrograms/h; groups 2, 4, and 6) in 10% ethanol-saline (vehicle), or vehicle alone (groups 1, 3, and 5), as a continuous infusion for 24 h beginning 7 days after insertion (s.c.) of blank (groups 1 and 2) or P4-containing (groups 3-6) implants. The effect of P4 withdrawal was assessed by removing P4-containing implants at the beginning of vehicle (group 5) or E2 (group 6) infusion. Gonadotrope responsiveness (increase in serum LH induced by 500 ng GnRH, i.v.) was assessed at the end of infusion. In a companion study, anterior pituitary tissue was collected at the end of the 24-h infusion period. Infusion of E2 increased (P < 0.05) GR relative to GR noted in control wethers receiving vehicle alone. The magnitude of E2-induced augmentation of GR was not affected by concurrent administration of P4 or P4 withdrawal. Pituitary tissue concentrations of GnRH receptor and GnRH receptor mRNA were significantly reduced in wethers implanted with P4. This P4-induced decrease in tissue concentration of GnRH receptor and GnRH receptor mRNA was not reversed during the 24-h period after P4 withdrawal. Steady-state concentrations of GnRH receptor and GnRH receptor mRNA were significantly increased by E2. However, the magnitude of oestrogen-induced increase in tissue concentrations of GnRH receptor mRNA was not significantly affected by P4 or P4 withdrawal. Conversely, concurrent progestin stimulation potentiated the E2-induced augmentation of tissue concentrations of GnRH receptor. However, this P4-induced potentiation of the oestrogenic response was not evident 24 h after removal of the P4-containing implants. Steady-state concentrations of mRNA encoding the LH beta and FSH beta subunits were reduced (P < 0.05) by P4. Infusion of E2 had a similar affect. These data indicate that prolonged progestin stimulation leads to a decrease in tissue concentrations of GnRH receptor and GnRH receptor mRNA. This P4-induced suppression of GnRH receptor activity is not reversed within 24 h of P4 withdrawal. In addition, the increase n steady-state concentrations of GnRH receptor and GnRH receptor mRNA induced by E2 is not compromised by concurrent progestin stimulation.