Microtubules and regulation of granulosa cell steroidogenesis by porcine granulosa cell conditioned medium.

Possible involvement of microtubules in the regulation of granulosa cell steroidogenesis by large follicle granulosa cell conditioned media (LGCCM) was assessed by monitoring the effect of agents that alter the cytoplasmic microtubule-tubulin equilibrium. The changes in microtubule organization and cell shape were examined by immunohistochemical procedure and morphometric analysis. Progesterone production stimulated by LGCCM was reduced by colchicine (agent that polymerizes microtubules) in a dose-dependent manner. In contrast, LGCCM stimulated progesterone secretion was significantly decreased by microtubule stabilizing agent (taxol: 1-10 microM). Cultured granulosa cells with a flattened appearance and projections after 24 h of incubation assumed a spherical configuration and were devoid of cytoplasmic processes when cultured with these agents. LGCCM stimulated GCs showed a reduction in the perimeter as compared with controls, although some cytoplasmic processes were observed. These findings suggest the involvement of microtubules in the regulation of cultured granulosa cell progesterone production by LGCCM, possibly through an effect on subcellular organelle distribution by altering the morphology of granulosa cells.