Abstract The contribution of the adrenal medulla and/or the sympathetic nerves to the plasma levels of Met-enkephalin was investigated. Rats were divided into four groups: sham-operated/saline, sham-operated/guanethidine, adrenal-demedullated/saline, demedulla-ted/guanethidine. After 4 weeks of injection with either saline or guanethidine (25 mg/kg/day), animals were cannulated in the left carotid artery for blood sampling. Three days later, blood samples were taken before and at 2 and 30 min of restraint stress. Adrenal demedullation lowered basal plasma epinephrine levels markedly and prevented entirely the increase induced by restraint stress. Chronic guanethidine treatment lowered basal plasma norepinephrine levels and decreased the response to stress. Guanethidine treatment increased the basal plasma epinephrine level without affecting the response to stress. The combination of guanethidine plus adrenal demedullation lowered basal plasma concentrations of all three catecholamines and further attenuated the norepinephrine response. Restraint stress increased plasma native and peptidase-derivable Met-enkephalin. Adrenal demedullation, resulting in greater than 95% depletion of adrenal catecholamines and significant depletion of adrenal Met-enkephalin, did not inhibit the stress-induced increase in plasma Met-enkephalin, and in fact, was associated with a potentiated response to stress. Guanethidine treatment with or without demedullation increased baseline plasma native Met-enkephalin and abolished the stress-induced increase in plasma native and peptidase-derivable Met-enkephalin. Thus, the stress-induced increase in plasma Metenkephalin results from release from sympathetic nerves, rather than adrenal medulla. However, the sympathetic nerves and adrenal medulla together do not appear to account entirely for basal concentrations of circulating Met-enkephalin. Hepatic portal vein plasma concentration of native Met-enkephalin was greater than that in the carotid artery, suggesting contribution from the gastrointestinal tract; however, evisceration did not decrease plasma native Met-enkephalin. Some compensatory mechanism results in elevation of basal plasma native Met-enkephalin in sympathectomized rats. Also, in the absence of the adrenal medulla there is a compensatory increase in the amount of Met-enkephalin released into the circulation in response to stress.