This study was designed to compare the effects of single and repeated administration of a discrete 2-min restraint stress on serotonin (5-HT) and catecholamine neuron activity in various regions of rat brain. A single 2-min restraint stress significantly increased the 5-hydroxyindoleacetic acid (5-HIAA) and 5-HT responses in hypothalamus and cerebral cortex and the 5-HIAA response in brainstem. A second 2-min restraint stress applied 90 min after the initial stress did not appreciably alter the steady-state concentrations of 5-HIAA and 5-HT nor did it produce any further changes in the 5-HIAA and 5-HT responses compared to those seen following a single stress in these 3 brain regions. In addition, the synthesis rate of 5-HT in anterior hypothalamus, posterior hypothalamus, hippocampus and brainstem was not altered by a second stress applied 90 min after the initial stress. In contrast, a second 2-min restraint stress applied 30 or 60 min after the initial stress significantly increased the 5-HIAA concentration in hypothalamus, cerebral cortex and brainstem. Also, the synthesis rate of 5-HT was greater following application of a second stress at 30 min than following either a single stress or a second stress applied at 90 min. Following application of a single 2-min restraint stress the hypothalamic concentration of norepinephrine (NE) was significantly decreased at 5 min after onset of the stress and returned to prestress levels by 15 min; the hypothalamic dopamine (DA) concentration was significantly increased at 30 min after the onset of the stress, while the hypothalamic epinephrine (EPI) concentration remained unchanged. A second 2-min restraint stress applied at 30 min markedly lowered NE concentrations in whole and mediobasal hypothalamus but not in laterobasal hypothalamus, and the NE concentrations remained decreased for a period lasting at least 60 min; there was a significant decrease in the hypothalamic EPI concentration 60 min after application of the second stress at 30 min. In addition, the synthesis rate of catecholamines was significantly greater in anterior but not in posterior hypothalamus after application of a second stress 30 min after the initial stress than following either a single stress or a second stress applied at 90 min. Negative correlations were demonstrated between increased synthesis rates of both hypothalamic 5-HT and anterior hypothalamic catecholamines and decreased corticosterone response to single and repeated stress.(ABSTRACT TRUNCATED AT 400 WORDS)