The sex steroid hormones that affect development in birds have been thought to be produced exclusively by the embryo or neonate. I used radioimmunoassay to measure the amounts of androstenedione, 5a-dihydrotestosterone, testosterone, 173-estradiol, and corticosterone in the yolk of freshly laid canary (Serinus canaria) and zebra finch (Poephilaguaata) eggs.Testosterone was found in both canary and zebra finch eggs, but its contents were much higher in the former than in the latter. The testosterone content of canary eggs in a same clutch increased with the order of laying, regardless of the genetic sex of the offspring that hatched from these eggs. Yolk testosterone was also present in the eggs of female canaries that were kept without a male, indicating that it is ofmaternal origin. The social rank of juvenile canaries was positively correlated with the concentration of yolk testosterone in the eggs from which they hatched, suggesting that the development of aggressive behavior of offspring might be subject to modification by maternal testosterone. These findings indicate that female songbirds can bestow upon their eggs a dose of hormone that modifies the behavior of offspring. Variable doses of these hormones might explain some of the individual variation in offspring behavior.Development of sex in higher vertebrates occurs in two steps. First the genotype determines the production of ovaries or testes. Then differential secretion of sex hormones determines the development of male and female secondary sex characters and behavior (1-4). Earlier studies of sex hormones in sexual differentiation (5-10) assumed that in birds, in which the embryo develops in the egg outside the mother, the developing organism was the main source of sex hormones that influence development. However, steroid hormones are lipophilic, and they may pass from the mother into the lipoprotein matrix of egg yolk during vitellogenesis. The presence of such hormones might affect developmental processes before embryonic steroidogenic tissue becomes active and could influence the fitness of offspring. That steroid hormones can pass from the female's circulation into the egg is suggested by experiments that show modification of offspring sexual differentiation in response to treatment of the egg-laying female with exogenous estradiol (11,12 removed, and a small yolk sample (10-15 mg) was obtained by inserting a 25-gauge needle through the small end of the egg into the yolk. After sealing the hole with transparent wound dressing, the eggs were returned to the nests, so that clutch size was not changed. The yolk sample was weighed, diluted with 0.5 ml of distilled water, mixed, and frozen for hormone assays. In addition, eggs from three females that were kept without a male were removed daily from the nests during the 1992 breeding season. In this case eggs were not returned to the nest, and the yolk was collected and frozen. Eggs laid at intervals of no more than 2 days were considered to belong to the same clutch.Steroid Hormone Assays. I...