The mechanism by which cysteamine reduces the PRL content of pituitary cells was studied in primary cultures of estradiol-induced pituitary tumors in Fischer 344 rats. The PRL content of these cells was effectively decreased by cysteamine, with an IC50 of 0.2 mM. Cells previously labeled with [3H]leucine were exposed to cysteamine (0.25 mM), and the intracellular content of [3H]PRL was measured by immunological or nonimmunological means, that is by immunoprecipitation and electrophoresis or by electrophoresis alone. The intracellular concentration of immunoreactive [3H]PRL was reduced by 53% by cysteamine, whereas [3H]PRL quantified by electrophoresis alone was not significantly affected. Our data indicate that cysteamine reduces the PRL content of pituitary tumor cells by causing the loss of its immunoreactivity.