Glucocorticoid receptors in bovine adrenal medullary cells in culture: regulation by cyclic nucleotides.
Glucocorticoid receptor levels within a given cell determine the glucocorticoid effect in the target tissue. Glucocorticoid receptors are present in adrenal medullary cells in culture where they are involved in the regulation of catecholamine biosynthesis. Modulation of glucocorticoid receptor protein and/or messenger RNA levels in response to cyclic nucleotides has been found in various cell types. In this study, we have investigated the effects of cyclic AMP and cyclic GMP on glucocorticoid receptor binding and glucocorticoid receptor-mediated function in Percoll-isolated bovine adrenal medullary cells in culture. Four-day treatment of cells with 8-bromo-cyclic AMP (10(-3) M) an analogue of cAMP, or forskolin (10(-5) M), an activator of adenylate cyclase, decreased soluble [3H]dexamethasone binding by 55 and 54%, respectively. 8-Bromo-cyclic GMP treatment decreased [3H]dexamethasone binding by 31 and 34% at 10(-5) and 10(-4) M, respectively. Treatment with 8-bromo-cyclic AMP or forskolin, but not 8-bromo-cyclic GMP, elevated cortisol levels in the medium of treated cells, presumably by elevating steroidogenesis in contaminating cortical cells. Cultures further purified to produce chromaffin-enriched cell cultures, also showed a loss (41%) in soluble [3H]dexamethasone binding when treated with 8-bromo-cyclic AMP (10(-3) M). Four-day treatment of standard Percoll-isolated cells with low concentrations of cortisol (10(-9) to 2 x 10(-7) M) similar to that found in the medium of 8-bromo-cyclic AMP-treated cells, did not decrease soluble [3H]dexamethasone binding, whereas higher cortisol concentrations (10(-6) M) produced a 62% loss in soluble binding. Adsorption of cortisol with bovine serum albumin (5 mg/ml) prevented a cortisol (10(-6) M)-induced loss in soluble [3H]dexamethasone binding with no effect on the 8-bromo-cyclic AMP-induced loss in binding, suggesting that the decrease in binding observed following 8-bromo-cyclic AMP treatment is not due to the release of cortisol from contaminating cortical cells. Finally, we report a loss in the ability of 8-bromo-cyclic AMP- or 8-bromo-cyclic GMP-treated cells to fully induce the activity of phenylethanolamine N-methyltransferase in response to cortisol, indicating that decreases in soluble [3H]dexamethasone binding translate into a decrease in the functional consequence of glucocorticoid receptor binding in adrenal medullary cells. In conclusion, these results indicate that long-term increases in cyclic nucleotide second messengers are able to decrease glucocorticoid receptor binding in bovine adrenal medullary cells, via a mechanism independent of released cortisol.(ABSTRACT TRUNCATED AT 400 WORDS)
Betito
K
K
McGill University, Department of Pharmacology and Therapeutics, Montreal, Quebec, Canada.
Diorio
J
J
Meaney
M J
MJ
Boksa
P
P
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000378:metabolism / Q000494:pharmacology; D000313:Adrenal Medulla / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002838:Chromaffin System / Q000166:cytology / Q000187:drug effects; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D003907:Dexamethasone / Q000378:metabolism; D006854:Hydrocortisone / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D009712:Nucleotides, Cyclic / Q000502:physiology*; D010625:Phenylethanolamine N-Methyltransferase / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism*
D009712:Nucleotides, Cyclic; D011965:Receptors, Glucocorticoid; D015124:8-Bromo Cyclic Adenosine Monophosphate; C016276:8-bromocyclic GMP; D003907:Dexamethasone; D010625:Phenylethanolamine N-Methyltransferase; D006152:Cyclic GMP; D006854:Hydrocortisone