5 alpha-Dihydrotestosterone (DHT) metabolism into 5 alpha-androstane-3 alpha, 17 beta-diol (alpha-diol) and 5 alpha-androstane-3 beta, 17 beta-diol (beta-diol) was studied in monolayer cultures of distinct cell populations from prepubertal male rats pituitaries. Cells were characterized through immunocytochemistry with the various antihormone antisera. Centrifugal elutriation was used to prepare a gonadotrope-enriched population "G" and a gonadotrope-depleted population "L", containing most lactotropes and somatotropes. Using centrifugation on Percoll gradient, two sub-populations, P1 and P2, were prepared by further fractionation of the "L" population. Cells were incubated for 48 h with [3H]DHT (1 microM, sp. act. 0.9 Ci/mmol) and metabolites extracted from the whole cell and medium. DHT was metabolized to about the same extent (30-40%) in all cell fractions. Compared with unfractionated population, the conversion of DHT into alpha-diol increased significantly in the P1 fraction, consisting of lactotropes, somatotropes and highly depleted in gonadotropes. This increase was lower in the somatotrope-enriched P2 fraction in which the amount of lactotropes was similar to P1 but that of gonadotropes slightly higher. In contrast, the conversion of DHT into alpha-diol decreased significantly in the "G" population compared with total or "L" fractions, whereas androstanedione formation, low in every population, increased significantly. The increase in alpha-diol formation could be related either to the decrease of gonadotropes or to a role of non-immunoreactive cells. As the beta-diol formation was constant in all cell types, the beta-diol/alpha-diol amount increased significantly in gonadotropes. Then, beta-diol and DHT could be both active steroids in gonadotrope regulation inasmuch as specific binding sites were identified for these two steroids. It can be concluded that DHT action at the pituitary level is subject to complex control mechanisms involving a specific balance of its metabolites in each particular cell type.