Neuropeptide-Y (NPY) and substance-P (SP), two peptides contained in the chromaffin granules of adrenal medullary cells, were found to partially inhibit both basal ACTH-stimulated release of aldosterone and 18-hydroxy-corticosterone by isolated rat zone-glomerulosa cells, without affecting the overall post-pregnenolone yield or basal progesterone output. Conversely, the exposure to both peptides increased 11-deoxy-corticosterone and corticosterone secretion. These data indicate that NPY and SP are able to exert a direct suppression of 18-hydroxylase activity in rat zona-glomerulosa cells, without conceivably altering the earlier steps of aldosterone synthesis. The possible physiological implications of these findings are discussed in light of previous studies suggesting a net adrenoglomerulotrophic effect of NPY and SP in vivo.