ANDROGEN BINDING IN PERIPHERAL-TISSUES OF FETAL RHESUS MACAQUES - EFFECTS OF ANDROGEN METABOLISM IN LIVER

In rhesus monkeys sexual differentiation of the brain and reproductive tract (RT) is androgen-dependent. Presumably these effects are mediated through the androgen receptor (AR). The AR has not been characterized in fetal tissues such as liver, kidney, heart, spinal cord and RT in this species. We characterized AR binding using [H-3]R1881 as the ligand in cytosols from tissues obtained on days 100-138 of gestation. Scatchard analyses revealed a single, saturable, high affinity AR in liver, kidney, heart, spinal cord and RT. The apparent dissociation constant (K(d)) ranged from 0.52 to 0.85 nM with no significant tissue differences. The number of AR (B(max); fmol/mg protein) differed significantly (P < 0.01) between tissues (liver > RT >> kidney greater-than-or-equal-to heart greater-than-or-equal-to spinal cord). Radioinert testosterone (T) and 5-alpha-dihydrotestosterone (DHT) but not androstenedione, progesterone, estradiol-17-beta, estrone or cortisol in a 50-fold molar excess inhibited [H-3]R1881 binding to the AR in spinal cord, heart, kidney and RT. However, in liver only DHT competed significantly (P < 0.01) for binding. This difference in binding of DHT vs T in the liver was further investigated by incubating liver and kidney cytosols with [H-3]DHT and [H-3]T at 4-degrees-C. We identified the metabolic products by mobility on Sephadex LH-20 columns and reverse isotope dilution. Liver cytosols metabolized [H-3]DHT to 5-alpha-androstane-3-alpha, 17-beta-diol (5-alpha-diol) and [H-3]T to 5-beta-androstane-3-alpha,17-beta-diol (5-beta-diol) at 4-degrees-C. In contrast, kidney cytosols metabolized [H-3]DHT while [H-3]T remained unchanged. Further studies indicated that a 50-fold molar excess of 5-alpha-diol inhibited the binding of [H-3]R1881 in liver cytosols by about 50% whereas the same molar concentration of 5-beta-diol had no effect. These data demonstrate the presence of AR in peripheral tissues of fetal rhesus monkeys and suggest that androgens through their receptors may affect development of these tissues. Liver cytosols are capable of metabolizing T and DHT at 4-degrees-C at conditions similar to those used for measuring cytosolic AR. However, T and DHT are metabolized differently, generating different isomers which have different affinities for hepatic AR.