The organ-culture technique was used to investigate the effects of lysomotropic agents (NH4Cl and chloroquine) and of modifiers of microfilaments (cytochalasin B) and microtubules (colchicine) on the induction of casein synthesis and the accumulation of casein mRNA by prolactin in the rabbit mammary gland. Neither chloroquine nor NH4Cl altered the lactogenic action of prolactin. Cytochalasin B attenuated the response to prolactin in terms of casein synthesis. However, this drug did not hamper the accumulation of casein mRNA. Colchicine exhibited a marked specific inhibitory effect on the induction of casein synthesis. It also prevented the accumulation of casein mRNA. These results suggest that a putative degradation of the internalized prolactin--receptor complex by lysosomes is not strictly involved in prolactin action. In addition, the integrity of the microfilaments seems unnecessary in the process of casein-gene activation by prolactin. By contrast, the integrity of the microtubule network seems absolutely necessary to ensure the transmission of prolactin information to the nucleus.