11beta-hydroxysteroid dehydrogenase is a predominant reductase in intact rat Leydig cells

11beta-Hydroxysteroid dehydrogenases (11beta-HSDs) interconvert active corticosterone and inert 11-dehydrocorticosterone. In tissue homogenates, 11beta-HSD type 1 (11beta-HSD-1) exhibits both 11beta-dehydrogenase (corticosterone inactivating) and 11beta-reductase (corticosterone regenerating) activities, whereas 11beta-HSD type 2 (11beta-HSD-2) is an exclusive dehydrogenase. In the rat testis, 11beta-HSD has been proposed to reduce glucocorticoid inhibition of testosterone production, promoting puberty and fertility. This hypothesis presupposes dehydrogenation predominates. 11beta-HSD-1 immunoreactivity has been localised to Leydig cells. However, recent studies suggest that 11beta-HSD-1 is predominantly an 11beta-reductase in many intact cells. We therefore examined the expression and reaction direction of 11beta-HSD isozymes in cultures of intact rat Leydig cells. Reverse transcriptase PCR demonstrated expression of 11beta-HSD-1, but not 11beta-HSD-2 mRNA in rat testis. Primary cultures of intact rat Leydig cells showed predominant 11beta-reductase activity, activating 50-70% of 11-dehydrocorticosterone to corticosterone over 3 h, whereas 11beta-dehydrogenation was <5%. Although both dexamethasone (10 nM) and corticosterone (1 microM) modestly inhibited LH-stimulated testosterone production by Leydig cells, inert 11-dehydrocorticosterone (1 microM) had similar effects, suggesting 11beta-reductase is functionally important. Carbenoxolone (10(-5) M) inhibited 11beta-reduction in intact Leydig cells. However, although carbenoxolone reduced Leydig cell testosterone production, this also occurred in the absence of glucocorticoids, suggesting effects distinct from modulation of corticosteroid access to Leydig cells. In conclusion, rat Leydig cell 11beta-HSD-1 is unlikely to reduce glucocorticoid access to testicular receptors. More likely, 11beta-reductase amplifies glucocorticoid action, perhaps to maintain Leydig cell metabolic and endocrine functions.