CONVERSION OF 4-ANDROSTENEDIOL AND 5-ANDROSTENEDIOL TO TESTOSTERONE, AND CONVERSION OF DEHYDROEPIANDROSTERONE TO 4-ANDROSTENEDIOL BY RAT TESTIS IN VITRO

Abstract

Whole testis tissue preparations of rat or man were incubated with androst-4-ene-3[beta], 17[beta]-diol(4ADL) (20 or 40[mu]g in one flask containing 1 or 2 testes) for 2 hr. in Krebs-Ringer bicarbonate buffer, pH 7.4, in 95 percent O2-5 percent CO2 gas phase. The major product derived was proven to be testosterone by means of paper and gas chro-matographic analyses and by derivative formation. Human chorionic gonadotropin (50 or 100 U/l flask) had a minor influence on the conversion of 4ADL to testosterone by rat testis in vitro. Androst-5-ene-3[beta], 17[beta], -diol(5ADL) was also converted to testosterone in the same time as that observed for 4ADL. Conversion of these 2 andro-stenedioles to androst-4-ene-3[beta], 17-dione(4ADN) was observed to minor extent compared, to the conversion of dehydroepiandrosterone (DHEA) to 4ADN. 4-C14-DHEA was incubated with rat testis tissue preparation for 2 hr., and 8 radioactive compounds converted from DHEA were separated on paper by extensive chromatography. Among these compounds, testosterone and 4ADN were identified by recrystal-lization with carrier steroid to constant specific activity. A compound behaving very similarly to the reference 4ADL on papers was separated. Oxidation of these compounds with chromium trioxide yielded 2 products which showed the same mobilities as 4ADN and testosterone. An aliquot of the compound was added with carrier 4ADL and recrystallization showed a nearly constant specific activity. The testosterone zone derived from the compound by oxidation with chromium trioxide was added with carrier testosterone for recrystallization, and the constant specific activity was demonstrated.