1 The aim of this study was to investigate the mechanism by which propylthiouracil (PTU) exerts its inhibitory e ects on the production of testosterone by rat testicular interstitial cells. 2 The plasma testosterone concentration was decreased 60 and 120 min after an intravenous infusion of PTU (10 or 20 mg kg 71 ), but the concentration of plasma T 4 was una ected by the drug treatment.3 Exposure of anterior pituitary tissue to PTU (3 ± 12 mM) in vitro did not a ect either basal or gonadotropin-releasing hormone (GnRH)-stimulated luteinizing hormone (LH) release. 4 PTU (3 ± 12 mM) inhibited both the basal and the human chorionic gonadotropin (hCG, 0.05 iu ml 71 )-stimulated release of testosterone from rat testicular tissue in vitro; at the highest concentration tested (12 mM), it also inhibited the forskolin or 8-bromo-adenosine 3':5'-cyclic monophosphate (8-Br-cyclic AMP)-stimulated release of testosterone. 5 The 25-OH-cholesterol (10 77 ± 10 75 M)-stimulated release of pregnenolone and testosterone by the testicular interstitial cells was inhibited by PTU (12 mM, P50.05). 6 The results suggest that the inhibitory actions of PTU on testosterone secretion are exerted, at least in part, at the testicular level through a mechanism which is independent of thyroid status and which involves a reduction in P450scc activity and, hence, in the conversion of cholesterol to pregnenolone.