Eight inbred strains of mice with varying incidences of spontaneous mammary tumor were compared in regard to prolactin and growth hormone concentrations in sera, pituitary glands and urine. Serum prolactin was compared under basal conditions as well as after stimulation with perphenazine. Both hormones were measured with specific, homologous radioimmunoassays. Although some strains having a high incidence of mammary tumors had high levels of prolactin in sera, urine and pituitary glands, neither basal nor perphenazine-induced serum concentrations showed a consistent pattern across mouse strains that correlated with the incidence of mammary tumors. Growth hormone levels in sera, pituitary glands and urine also had no characteristic pattern that applied to all strains studied. The ratio of prolactin depleted from the pituitary gland to prolactin detected in serum after perphenazine injection, which reflected the metabolic clearance rate of prolactin, was highest in two strains with a high incidence of mammary tumors and relatively lower in low-tumor strains. These results suggest that if prolactin plays a part in mammary tumor development in mice, its mechanism varies with strains: while hyperprolactinemia may be the means in some strains, a peculiarity in the metabolism of the hormone may be more important in others.