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We used radioimmunoassay (RIA) to measure monthly serum levels of unconjugated and conjugated sex steroids (testosterone T, androstenedione A, estradiol E(2), and estrone E(1)) in 4 male and 4 female foals during their first year of life. Maximal production of sex steroids was detected from April to August with hormonal peaks, corresponding to the natural breeding season in adults. In males, only A levels were more steady. Total estrogens (unconjugated plus conjugated E(2) and E(1)) were the major steroids in immature males in contrast to adults. Estrogens generally peaked in young females before males; the major estrogen was E(1), and total estrogens overtook total androgens (unconjugated and conjugated T and unconjugated A). We also sampled 3 male and 3 female foals with bone alterations in adulthood. For all animals, serum levels of four bone formation markers were obtained: osteocalcin (O), hydroxyproline (HP), and alkaline phosphatase (AP), and a radiographic score was determined. Only male foals with normal skeletal frame (good radiographic score GRS) in adulthood showed a correlation (P < 0.01) between the distribution frequency of each bone formation marker and unconjugated E(2) or E(1) levels; this finding highlighted the role of unconjugated estrogens in bone maturation in horses, since this was not found in the groups with bone alterations. In females, the threshold of estrogen synthesis and sensitivity was probably sufficient to be a nonlimiting factor at this stage of development. Our results strongly suggest a differential regulation of the estrogen/androgen balance in horses according to sex, sexual maturation, and photoperiod. Moreover, estrogens appear to be crucial for skeletal development in male colts, and these steroids are good modulators of skeletal frame characteristics in adulthood.