Objective: To determine CA-125 levels in cervical mucus (CM) during the menstrual cycle and their relationship to gonadal steroids and ovulation. Design: Prospective study. Setting: Two academic tertiary referral centers. Participants: Thirteen women with a normal fertility work-up. Interventions: CA-125 and protein concentrations were measured in CM aspirated from the endocervical canal on alternate days in the early follicular and luteal phases and on a daily basis during the periovulatory period. Results were correlated with hormonal determinations, serum CA-125 levels, and ultrasound examination. Results: Twenty ovulatory nonconceptional cycles were analyzed. Although the mean (+/-SD) concentration of CA-125 in CM (173,900 +/- 128,900 arbitrary U/mL) appeared relatively constant along the cycle, a large variation among the different samples was observed, ranging from 9,000 to 830,000 arbitrary U/mL. No clear trend could be detected as related to hormonal changes and ovulation. However, when the mucus CA-125 concentration was multiplied by the total volume of the correspondent sample, a clear periovulatory increase of total CA-125 levels was found. This was further supported by a similar trend showed by the calculated CA-125:protein concentration ratio. Conclusions: CA-125 is present in CM in high concentrations that vary widely along the cycle. Although no cyclical variation in CA-125 concentration could be determined, there was an apparent increase of total CA-125 levels parallel to the augmented mucus production during the periovulatory period. This further suggests a possible involvement of this glycoprotein in the secretory process of endocervical glands.