Exposure of an organism to any of a variety of stressors markedly activates the sympathoadrenal and hypothalamic-pituitary-adrenocortical systems. Interactions of these major stress systems occur at several levels in the periphery and the brain. In the present study, we used sham-operated or adrenalectomized cortisol-treated conscious rats to examine glucocorticoid effects on indices of CA release, metabolism, and synthesis, and on CA biosynthetic enzyme activities and gene expression at baseline and during immobilization stress (IMO). Adrenalectomy (ADX) stimulated basal and stress-induced increments in norepinephrine release, reuptake, metabolism, turnover, and biosynthesis. Loss of adrenomedullary hormones after ADX did not appear to contribute to these increments. Cortisol treatment reversed the ADX effects on CA indices and suppressed catecholaminergic responses to IMO in intact rats. These results suggest that endogenous glucocorticoids restrain responses of catecholamine turnover, synthesis, release, reuptake, and metabolism during stress. In contrast, in intact rats, continuous administration of cortisol lasting for 7 days exaggerated the IMO-induced increases in plasma CA levels. Inhibition of DOPA conversion to dopamine elevated plasma DOPA levels in chronically cortisol-treated stressed rats compared to saline-treated ones, suggesting a cortisol-induced increase in tyrosine hydroxylation. Stress increases TH and PNMT activities and mRNA levels in the adrenal medulla. Hypophysectomy reduced adrenal PNMT but not TH mRNA levels in control and IMO rats. Pretreatment of hypophysectomized animals with ACTH fully restored the control and IMO-induced adrenal PNMT mRNA levels and augmented PNMT but not TH mRNA responses in intact rats. Long-term cortisol administration to intact rats also elevated adrenal PNMT but not TH mRNA levels. The results indicate a suppressive effect of endogenous glucocorticoids and a stimulatory effect of chronically elevated glucocorticoid levels on sympathoadrenal activity during stress. The results also suggest that a nonneuronal, nonpituitary factor contributes to TH gene expression during some forms of stress, whereas pituitary-adrenocortical factors play the essential role in the regulation of PNMT gene expression.