Effects of lysosomotropic agents and drugs which disrupt the cytoskeleton on prolactin receptor levels were studied in organ culture of rabbit mammary glands. As we have shown previously, prolactin is able to induce a down-regulation of its own receptor levels. In the present experiments, lysosomotropic agents, chloroquine, ammonium chloride and methylamine, in the presence of prolactin are capable of almost completely preventing this down-regulation. Interestingly, these lysosomotropic agents alone could increase the level of prolactin receptors, which confirms that the degradation of prolactin receptors occurs in the lysosomes and that this is a rapid process which can proceed in the absence of prolactin. The almost complete lack of effect of microfilament (cytochalasin B) or microtubule (colchicine) disrupting drugs seems to indicate that the cytoskeleton is not involved in the down-regulation process.