The influence of androgenic status on basal and stimulated cAMP production, adenylyl cyclase activities and immunoblot quantified GS alpha and Gi alpha 2 subunits of the adenylyl cyclase regulatory proteins were compared in confluent preadipocytes from subcutaneous (SC) and deep-intraabdominal (epididymal) fat deposits. Maximal cAMP response to isoproterenol was lower in SC than in epididymal preadipocytes. After castration, this site-specific difference was suppressed. cAMP response to 2-chloroadenosine, which was identical in the two types of preadipocytes, was decreased by castration in epididymal cells but not in SC cells. The catalytic activity of adenylyl cyclase and its maximal response to GTP were higher in epididymal than in SC preadipocytes. This response to GTP was decreased by castration in epididymal preadipocytes while it remained unchanged in SC preadipocytes. The catalytic activity of adenylyl cyclase was unchanged by androgenic status whatever the cell localization. Levels of GS alpha quantified by immunoblotting were not modified whatever the androgenic status and cell origin. Levels of Gi alpha 2 were not affected by the androgenic status as well, but were lower in SC than in epididymal cells. This study shows that components of the adenylyl cyclase system in preadipocytes are differently regulated by the androgenic status depending on the anatomical origin of the cells.