The central release of vasopressin was studied in anesthetized rats using push-pull perfusions and radioimmunoassay of the hormone. A basal release was observed in the lateral septum and in the lateral ventricle, whereas no vasopressin was detected in the perfusates from the caudate nucleus. Under osmotic stimulation, vasopressin release increased up to 12 and 60 times basal levels following i.p. injections of 5 ml and 10 ml/kg b.wt. of 2 M NaCl, respectively. This increase was blocked by using a calcium-free perfusion medium containing 0.1 mM EGTA. In the lateral ventricle, osmotic stimulation (5 ml/kg of 2 M NaCl i.p.) had the same effect as in the septum. In the caudate nucleus, no release was observed. Hemorrhage also increased the septal release of vasopressin in 5 out of 6 animals tested. Electrical stimulation of the pituitary stalk and of the supraoptic nucleus was used to evoke the release of vasopressin into the bloodstream. Septal release slightly decreased during pituitary stalk stimulation, whereas it did increase during stimulation of the supraoptic region. Our results show that systemic stimuli for vasopressin release evoke both a peripheral and a septal release of the hormone. The dissociation of the effects of electrical stimulation of the pituitary stalk and of the supraoptic nucleus suggests, however, that the vasopressinergic neurones responsible for septal release are distinct from those which project to the neurohypophysis.