Preliminary observations of bovine adrenal fasciculata-reticularis cells in monolayer culture: steroidogenesis, effect of ACTH and cyclic AMP.
PMID 184635 · Acta endocrinologica · 1976 · DOI 10.1530/acta.0.0830373
Authors: C G Goddyer; J S Torday; B T Smith; C J Giroud
Abstract
Bovine adrenocortical cells dispersed by trypsin digestion of fasciculata-reticularis minces were maintained in monolayer culture for up to 6 weeks. During the first week cells grown in medium containing ACTH (1 mU/ml) secreted steroids at a rate 10 to 20-fold greater than control cultures, cortisol accounting for 80-90% of the corticotrophic response. Using tracer amounts of [3H] progesterone and [3H] pregneolone, the major products were cortisol, corticosterone, 11-deoxycortisol and 11-deoxycorticosterone in decreasing order of magnitude. After 10 to 15 days in culture steroidogenesis was no longer enhanced by ACTH. This was concomitant with an apparent loss of 11 beta-hydroxylase activity which was mainly manifested by a sharp increase in the formation of 44-deoxycortisol. Short-term incubations of these cells during the first week in culture provided evidence that steroidogenesis was related to ACTH concentrations (from 0.1 to 100 muU/ml) and stimulated by dibutyryl cyclic AMP, the corticotrophic responses being further enhanced by theophylline (0.5to 50 mumoles/5 ml). Exposure of the cells to ACTH (50 muU/ml) resulted in a rapid increase in intracellular cyclic AMP contractions concomitant with a progressive increase in the corticosteroids released into the medium.
Adrenal Cortex; Adrenal Glands; Adrenocorticotropic Hormone; Animals; Biological Assay; Bucladesine; Cattle; Cells, Cultured; Corticosterone; Cortodoxone; Culture Media; Cyclic AMP; Glucocorticoids; Hydrocortisone; Pregnenolone; Progesterone; Time Factors
Cite
C G Goddyer; J S Torday; B T Smith; C J Giroud. Preliminary observations of bovine adrenal fasciculata-reticularis cells in monolayer culture: steroidogenesis, effect of ACTH and cyclic AMP. Acta endocrinologica (1976). PMID 184635. DOI 10.1530/acta.0.0830373.