The indirect immunofluorescence technique was used to examine the pharmacology associated with reserpineinduced alterations in vasopressin and neurophysin (VP/NP) immunoreactivity in the external layer of the median eminence in the rat. Twenty-four hours after injection of reserpine, a selective, marked depletion of VP/NP immunoreactivity from the external layer is apparent. Pretreatment with the monoamine oxidase inhibitors, pargyline and tranylcypromine, prevents the depleting effect of reserpine, indicating that the acute effect of reserpine is mediated by monoamines. Acute intraventricular treatment with 6-hydroxydopamine, but not 5, 7-dihydroxytryptamine, mimicked the reserpineeffect, suggesting that catecholamines mediate reserpine depletion of VP/NP immunoreactivity from the external layer. The experimental results are consistent with a regulatory model in which catecholamines tonically inhibit VP/NP release from terminals in the external layer of the median eminence. Although the studies do not definitively determine the functional relationship between VP and ACTH, the anatomical location of these terminals, the dramatic change in the VP/NP content of these terminals in response to reserpine, and the lack of a response to dehydration suggest that this pool of vasopressin may contribute to ACTH hypersecretion in response to reserpine.