INTRAADRENAL STEROID-METABOLISM IN THE GUINEA-PIG - GUINEA-PIG ADRENAL MICROSOMES METABOLIZE ANDROSTENEDIONE IN A MANNER DISTINCT FROM LIVER-MICROSOMES

Several immunochemical homologs of hepatic cytochromes P450 (CYPs) capable of steroid catabolism have been identified in the guinea pig adrenal cortex. Their predominance in males suggests a role in sex-differentiated metabolism of androgens. Therefore, we examined the ability of microsomes from male guinea pig adrenals and liver to metabolize androstenedione. Microsomes were incubated in the presence of radiolabeled steroids, the products of the reaction extracted, separated by TLC, and visualized by autoradiography. Metabolites were identified by comigration with commercially available standards in several solvent systems, in one and two dimensional TLC. Microsomes from both tissues metabolized androstenedione. However, the products formed differed markedly in the two tissues. Liver microsomes formed one major metabolite, testosterone. It represented 85% of the metabolized androstenedione. 6 beta-Hydroxylated androstenedione and testosterone each comprised 3-4% of the liver metabolites. In addition, at least 10 other products were formed, but taken together they constituted less than 8% of the metabolized androdostenedione. Adrenal microsomes, on the other hand, produced several major metabolites: 16 alpha-, 16 beta-, and 6 beta-hydroxyandrostenedione, plus one unidentified product constituted 93% of the metabolized androstenedione. 16 alpha-Hydroxylation of androstenedione was 60 fold, 16 beta-hydroxylation 12 fold, and 6 beta-hydroxylation 2.5 fold greater in adrenal than in liver microsomes. The unidentified product, which was the least polar, was formed exclusively by adrenal microsomes. The hydroxylation reactions perfomed by adrenal tissue are consistent with the presence in adrenal microsomes of immunochemical homologues of members of the CYP1A, 2B, 2C and 3A families which have known steroid hydroxylation functions in liver. The Kms of the formation of 16 alpha-, 16 beta- and 6 beta-hydroxyandrostenedione by adrenal microsomes are in the range reported for steroid hydroxylation reactions in rat liver tissue. Their distinct values suggest that these hydroxylation steps are performed by different CYPs. However, assignment of site-specific steroid hydroylation reactions to individual hepatic CYP homologs in the adrenal requires further investigation and is being pursued using combined techniques of biochemistry and molecular biology.