The objective of this study was to determine whether concentrations of hypophyseal and serum insulin-like growth factor-I (IGF-I) and binding activities of serum and hypophyseal IGF-binding proteins (IGFBPs) differ with stage of the estrous cycle in mature beef cows. Cows were assigned to the following Stages of the estrous cycle based on serum concentrations of progesterone (P-4,) and ovarian structures at death: days 1-5 (day 0 = estrus; n = 18), days 6-10 (n = 24), days 11-16 (n = 39), and days 17-21 (n = 7). Serum samples collected at death and anterior pituitary homogenates were analyzed for IGF-I, LH, and FSH by RIA. Serum and pituitary IGFBPs were evaluated by ligand and immunoblot analyses. Serum samples contained IGFBP activity at 44 and 40 kilodaltons (kDa; IGFBP-S), 34 kDa (IGFBP-2), 30 kDa (IGFBP-5), 28 kDa, and 24 kDa. The intensity of binding by the different sized proteins in serum remained constant throughout the estrous cycle. IGFBPs detected in anterior pituitaries included a 36-/40-kDa doublet (IGFBP-3), a 32-kDa protein (IGFBP-2), and a 29-kDa IGFBP (IGFBP-5). The intensity of [I-125]IGF-I binding to IGFBPs was greater (P < 0.05) during days 11-16 than days 1-5 or days 6-10 of the estrous cycle and was intermediate (P > 0.05) during days 17-21. Serum concentrations of LH were low(<0.4 ng/ml) during days 1-16 of the estrous cycle, but increased (P < 0.05) approximately 3- to 4-fold during days 17-21. LH concentrations in the anterior pituitary increased (P < 0.05) from the postovulatory period (548 +/- 52 mu g/g; days 1-5) to the late luteal phase (791 +/- 39 mu g/g; days 11-16) and were intermediate (P > 0.05) during the preovulatory phase (707 +/- 85 mu g/g; days 17-21). Concentrations of IGF-I and FSH in the anterior pituitary and serum did not differ (P > 0.05) by stage of the estrous cycle. A positive correlation among the different IGFBPs in the pituitary (P < 0.01) and between each pituitary IGFBP and serum P-4 (P < 0.05) existed. The significance (P < 0.0002) of the correlation between pituitary IGFBP-3 and P-4 was greater (P < 0.02) than that between the other pituitary IGFBPs and P-4. In summary, IGFBP activity in the anterior pituitary, but not that in serum, changed with stage of the estrous cycle in association with serum concentrations of P-4. The absence of similar changes in serum or anterior pituitary concentrations of IGF-I supports the hypothesis that IGFBPs may regulate the actions of IGF-I on gonadotropin release from the anterior pituitary gland during the estrous cycle of beef cattle.