A lack of pituitary LH stores has been implicated as the cause of seasonal anestrus and failure to ovulate during the spring transition period in mares. In this experiment, 40 mares were used to study the effects of GnRH, estrogen, and an estrogen-GnRH combination on increasing releasable pituitary LH. Mares were stratified based on their ability to secrete LH in response to a 950-micrograms challenge of GnRH (n = 10 per group) and then assigned to one of four treatment groups: 1) controls, given no treatment; 2) 1 mg of estradiol-17 beta in oil i.m. daily for 8 d; 3) 200 micrograms of GnRH analogue des-Gly10, [D-ala6]-LHRH ethylamide in saline i.m. twice daily for 8 d; or 4) estradiol for 4 d then estradiol plus GnRH for four subsequent days. Blood was collected on d 1, 3, 5, and 7 of treatment, and serum was assayed for LH. On d 10 after initiation of treatment, mares were again challenged with GnRH (950 micrograms), and blood was collected for 4 h. Concentrations of serum LH did not vary significantly in control, estradiol-treated, or estradiol plus GnRH-treated mares among treatment days. In contrast, administration of GnRH alone increased (P < .05) concentrations of LH on d 5 and 7. Response to GnRH challenge, as measured by area under the LH curve (AUC) and peak LH, was greater (P < .05) for mares administered GnRH (7,307.1, 67.6 ng/mL, respectively) and GnRH plus estradiol (5,691.4, 60.3 ng/mL) than for mares given estradiol alone (1,519.4, 22.1 ng/mL) or no treatment (1,213.8, 19.4 ng/mL.(ABSTRACT TRUNCATED AT 250 WORDS)