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Neuropeptides (NP) B and W are hypothalamic peptides involved in the regulation of feeding and neuro-endocrine axes. Evidence has been provided that NPB and NPW act on both the central and the peripheral branches of the rat hypothalamic-pituitary-adrenocortical axis, and we carried out in vivo and in vitro studies to gain insight into this topic. Reverse transcription-polymerase chain reaction showed the expression of NPB, NPW and their receptors in both adrenal cortex (zonae glomerulosa and fasciculata-reticularis) and adrenal medulla, where immunocytochemistry also detected the presence of abundant NPB- and NPW-immunoreactivity. The acute subcutaneous administration of NPB (0.5 or 1.5 nmol/100 g) did not alter ACTH plasma concentration, while that of NPW (1.5 nmol/100 g) decreased it. Neither NPB nor NPW affected the blood level of aldosterone, while both peptides (0.5 nmol/100 g) raised that of corticosterone. NPB (10(-6) M) lowered ACTH-stimulated aldosterone secretion, and basal and ACTH-stimulated corticosterone production from adrenal quarters containing both cortical and medullary tissues. NPW (10(-6) M) enhanced basal aldosterone secretion from adrenal quarters, and the effect was suppressed by the beta-adrenoceptor antagonist l-alprenolol (10(-5) M). NPW did not affect corticosterone production. Collectively, our findings allow us to draw the following tentative conclusions: i) ACTH-independent extra-adrenal mechanism(s) are operative in vivo, by which NPB and NPW stimulate adrenal glucocorticoid, but not mineralocorticoid secretion; ii) in vitro the interaction of NPB with adrenal medulla activates unknown mechanism(s) hampering adrenocortical steroidogenic machinery; and iii) NPW stimulates in vitro aldosterone secretion by enhancing the release of medullary catecholamines, which in turn activate beta-adrenoceptors located on zona glomerulosa cells.