Further characterization of the steroidogenic responsiveness of purified zona fasciculata/reticularis cells from bovine adrenal cortex before and after primary culture: Changing responsiveness to phosphoinositidase C agonists

I. M. Bird, C. D. Clyne, E. R.T. Lightly, B. C. Williams, S. W. Walker

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When bovine adrenocortical cells from the zona fasciculata/reticularis (zfr) are maintained in primary culture, cortisol secretion in response to acute stimulation with ACTH and adrenaline (which activate adenylate cyclase) is seen to increase steadily over the first 48 h, while secretion in response to angiotensin II and acetylcholine (which activate phosphoinositidase C) shows an initial decline in the first 24 h and a recovery to maximum after 48 h. We have investigated whether these discrepant changes in cortisol secretory response to the different agonists are due to changes in formation of the associated second messengers (cAMP or inositol phosphates), or altered coupling of these second messenger signals to steroid secretion. Increases in steroid secretion in response to ACTH and adrenaline were paralleled by increased cAMP. Steroid secretion in response to exogenous 8-bromoadenosine 3':5'-cyclic monophosphate also increased steadily during this 48-h period. Thus increased responsiveness was due to both increased second messenger formation and increased coupling to the steroid secretory response. The decreased steroid secretory response to angiotensin and acetylcholine after 24 h, and subsequent recovery after 48 h in culture, were accompanied by an increased formation of phosphoinositols after 24 h and a further increase by 48 h. However, the steroid secretory response to a combination of calcium ionophore and the protein kinase C activator, phorbol 12-myristate 13-acetate, was reduced after 24 hand recovered by 48 h of culture. Fura-2-loaded cells also showed an increase in intracellular [Ca2+] after 24 h in culture. Thus the impaired steroid secretory response to angiotensin II and acetylcholine after 24 h of culture was not due to reduced formation of second messengers but to a failure of Ca2+ and diacylglycerol so formed to activate the steroid secretory process. Reversible uncoupling of the steroid secretory response from the Ca2+ - and diacylglycerol-based but not the cAMP-based second messengers observed in bovine zfr cells suggests that differential control of steroid secretion and other cell functions may be possible in vivo for activators of phosphoinositidase C, and may explain apparently discrepant results from studies on other in-vitro adrenocortical cell preparations.

Original languageEnglish
Pages (from-to)21-28
Number of pages8
JournalJournal of Endocrinology
Issue number1
Publication statusPublished - 1 Jan 1992

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