Characterization of the steroidogenic responsiveness and ultrastructure of purified zona fasciculata/reticularis cells from bovine adrenal cortex before and after primary culture

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Abstract
Analysis by electron microscopy indicated that after 3 days of primary culture, purified bovine adrenal zonal fasciculata/reticularis (ZF/ZR) cells showed improved integrity of their ultrastructure, with an increased density of lipid droplets and smooth endoplasmic reticulum. The basal cortisol output was significantly (P < 0·05) greater on day 3 of culture than for the freshly isolated cells in six out of seven experiments. Similarly, in six experiments with ACTH (1 nmol/l) and five experiments with angiotensin II (10 nmol/l), the stimulated cortisol secretion was significantly (P < 0·01 for all 11 experiments) higher on day 3 of culture than in freshly isolated cells. No significant increase in cortisol secretion above basal was observed with noradrenaline at any concentration in the freshly isolated cells, whereas a dose-dependent increase in cortisol secretion was observed on day 3 of culture in all of four experiments. These findings were supported by cyclic (c) AMP output measured in one such experiment. Thus the basal cAMP output and that stimulated by ACTH (1 nmol/l) were significantly higher after culture (P < 0·001, n = five wells for basal comparison; P < 0·05, n = three wells for ACTH at 1 nmol/l). In agreement with the cortisol results, cAMP production was unaffected by any concentration of noradrenaline in the freshly isolated cells, whereas a dose-dependent rise was found after culture. Angiotensin II at all concentrations had no effect on cAMP production in freshly isolated or cultured cells. These studies demonstrate that the primary culture of purified bovine ZF/ZR cells increases basal steroidogenesis and leads to an enhanced responsiveness to the physiological stimuli, ACTH and angiotensin II. In addition, primary culture of purified bovine ZF/ZR cells revealed a noradrenaline response which was not readily observed in freshly isolated, purified ZF/ZR cells. Journal of Endocrinology (1989) 121, 317–324

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