This study was performed to investigate the effects of 17 alpha-ethinyl estradiol, a potent hypocholesterolemic agent at pharmacological doses, on cholesterol balance in the liver and extrahepatic tissues of the rat in vivo. Female Sprague-Dawley rats were treated with 17 alpha-ethinyl estradiol (5 mg/kg per day s.c. for 5 days) or with 4-aminopyrazolo(3,4-d) pyrimidine (20 mg/kg per day i.p. for 3 days). Both drug regimens suppressed plasma total and low density lipoprotein-cholesterol by more than 80%. Analysis of the kinetic parameters of low density lipoprotein transport did not show increased receptor activity in extrahepatic tissues during either treatment. 17 alpha-Ethinyl estradiol significantly increased low density lipoprotein tissue spaces and clearance rates in the liver, with a 5-fold increase in low density lipoprotein-receptor activity, whereas 4-aminopyrazolo(3,4-d)pyrimidine suppressed hepatic transport of low density lipoprotein probably due to a nospecific toxic effect. Treatment with 17 alpha-ethinyl estradiol markedly enhanced the hepatic expression of low density lipoprotein-receptor protein and mRNA despite a 7-fold increase in hepatic cholesteryl ester levels. Finally, treatment with both drugs increased cholesterol synthesis in several extrahepatic tissues, such as adrenals, ovaries, small bowel, and spleen. These findings confirm that 17 alpha-ethinyl estradiol at pharmacological doses markedly increases synthesis and expression of low density lipoprotein-receptor in the liver. Hypocholesterolemia, whether induced by activation of low density lipoprotein-receptors or by other mechanisms, fails to up-regulate low density lipoprotein transport in extrahepatic tissues, which rather respond by increasing local sterol synthesis. This suggests the occurrence of separate regulatory mechanisms for low density lipoprotein transport and cholesterol synthesis.