The involvement of microtubules (MT) and microfilaments (MF) in LH-regulation of luteal cell steroidogenesis was assessed at the middle stage of corpus luteum development. The influence of microtubule- and microfilament-altering agents on basal and LH-stimulated progesterone (P4) production and secretion into the incubation medium was determined by RIA. LH-stimulated P4 production was 2.5 times higher than in the control cultures. Cytochalasin B (Cyt B) was without effect on basal P4 synthesis but increased the basal fraction of P4 secreted into the incubation medium, while colchicine (Col) increased both basal P4 synthesis and the fraction of P4 secreted into the incubation medium. LH-stimulated progesterone synthesis was reduced by Col, but the fraction secreted into the incubation medium increased. Cyt B had no effect on LH-stimulated synthesis but it decreased the fraction of P4 secreted into the incubation medium. Our findings demonstrate significant differences in the effect of Cyt B and Col on steroidogenesis in corpus luteum. We conclude that microtubules play an important role in the process of LH-stimulated P4 synthesis, while microfilaments act in the process of basal and LH-stimulated P4 secretion.