HCG + ACTH Stimulation of in vitro dehydroepiandrosterone production in human fetal adrenals from precursor cholesterol and Δ-pregnenolone

Abstract

Fetal adrenal glands were obtained from legal abortions in the 14--22 gestational weeks. The adrenal cortex was separated from the medulla using a micromanipulator. The cortex was homogenized in a glass-teflon homogenizer and the microsomal fraction was isolated by centrifugation. Tissue slices were also prepared by the method of Deutsch. The microsomal fraction or the slices were incubated with (4-14C) pregnenolone or (4-14C) cholesterol in the presence of an NADPH regenerating system and oxygen. ACTH or HCG was added. After extraction and paperchromatography radioactivity was determined in a scanner. The conversion of pregnenolone to 17 alpha-hydroxypregnenolone and dehydroepiandrosterone was increased by ACTH. In the presence of HCG dehydroepiandrosterone production only was increased,--and this occurred in slices only. In the microsomal fraction HCG was without effect on steroid biogenesis from dehydroepiandrosterone.