The effects of estradiol feedback on pituitary luteinizing hormone (LH) content, serum LH concentration, and in vitro-translated alpha subunit was examined in the ewe. Three animal models were used representing positive, negative, and no estradiol feedback. Two experiments were carried out: (i) anestrous ewes were treated acutely with five Silastic estradiol implants to induce a LH surge (positive feedback) and (ii) ovariectomized ewes were treated chronically with an estradiol implant (negative feedback) or were not treated (no feedback). Pituitary RNA was prepared and translated in a cell-free system; the alpha subunit was identified by immunoprecipitation and NaDodSO4/PAGE. cpm/microgram of RNA and immunoprecipitated growth hormone and prolactin were used to evaluate possible differences in RNA translational efficiencies among the treatment groups. In experiment 1, significantly higher amounts of the alpha subunit were observed in animals exhibiting an estradiol-induced LH surge than in normal anestrous ewes (P less than 0.001). Examination of values from individual animals suggested a correlation between the stage of the LH surge, pituitary LH, and translated alpha subunit. In experiment 2, the amount of alpha subunit observed in animals exposed to chronic estradiol negative feedback was significantly less (P less than 0.005) than that in the untreated ovariectomized animals (no feedback) and no different from that in intact anestrous ewes. These results suggest that both the negative and the positive feedback effects of estradiol include regulation of the amount of alpha-subunit mRNA.