A model for studying regulation of aldosterone secretion: Freshly isolated cells or cultured cells?

Practically all studies relating to zona glomerulosa function have been performed either with freshly isolated cells or with cells used after 2 0r 3 days in culture. This study compares the step-by-step response (binding, second messenger production and aldosterone response) of isolated glomerulosa cells vs cells maintained in primary culture to the main stimuli of aldosterone secretion. One day in culture induces a decrease of 77 and 65% in the basal level of corticosterone and aldosterone secretions, compared to that observed in freshly isolated cells. In these conditions, the cells become more sensitive to most of their stimuli, but not all: e.g. important differences are noted in the dose-response of aldosterone secretion to adrenocorticotropin (ACTH), which is often shifted to a lower concentration sensitivity in cultured cells. For example, 0.1 nM ACTH stimulates steroid secretion by three-fold in isolated cells while 1 pM ACTH already induces a 25 and nine-fold increase, respectively, in corticosterone and aldosterone output in cultured cells. Moreover, some stimuli such as isoproterenol do not have any effect in isolated cells but do stimulate steroid secretion in cultured cells. In contrast, other stimuli, such as serotonin or DA (via DA 2 receptors) act preferentially in freshly isolated cells. The main observation derived from this study is that glomerulosa cells, under appropriate conditions, are able to respond to their main secretagogues even after 4 days in culture. At this time, glomerulosa cells maintain their ultrastructural characteristics and functional properties and, aside from a few exceptions, demonstrate higher sensitivity to their known stimuli. Culture conditions used in the past 5 years have helped demonstrate the regulatory role of a number of peptides which probably act via paracrine or autocrine pathways. However, the effects of certain stimuli must be studied in freshly isolated conditions since their respective receptor binding sites are lost in culture. In conclusion, comparative studies between isolated and cultured conditions must be undertaken before pursuing studies on either the mechanisms of action or interactions between newly identified regulators of aldosterone secretion.