The changes in alpha adrenoceptors in vasa deferentia of reserpinized rats in organ culture were examined by binding studies with 3H-clonidine, 3H-yohimbine and 3H-prazosin. On organ culture for 1 day, the 3H-clonidine binding sites decreased by 1.07 pmol/g tissue and after 2 days no binding sites were detectable. 3H-Yohimbine binding sites also decreased during culture, but in smaller extent than 3H-clonidine binding sites. On the other hand, 3H-prazosin binding sites showed no apparent change in amount during culture. Addition of 0.1 mM hydrocortisone or dexamethasone to the culture medium inhibited the decrease in 3H-clonidine binding sites (1.7 pmol/g tissue), and the effect of glucocorticoid was blocked by the inhibitors of protein synthesis, cycloheximide and puromycin. However, hydrocortisone showed no effect on the 3H-yohimbine and the 3H-prazosin binding sites. An anti-endocytotic agent, an anti-microtubular agent and protease inhibitors had no effects on the decrease of 3H-clonidine binding sites during culture. These results suggest that the amount of alpha-2 adrenoceptors can change rapidly while alpha-1 adrenoceptors are stable and that glucocorticoids are important in regulation of conformation of alpha-2 adrenoceptor through synthesis of certain protein(s).