The mechanism and routes through which peptide tyrosine-tyrosine (PYY) exerts its anorectic effects are still largely unknown. In the present study, we investigated the roles of the area postrema (AP), subfornical organ (SFO) and vagus nerve in mediating the anorectic effect of PYY using PYY3-36 conjugated to human serum albumin (PYY3-36-HSA) in rats. PYY3-36-HSA is a large molecule that does not penetrate the blood–brain barrier, and thus provides a useful tool to discriminate between the central (brain) and peripheral actions of this peptide. PYY3-36-HSA induced significant reductions in food and body weight gain up to 24 h after administration. The anorectic effect of PYY3-36-HSA was delayed for 2 h in rats in which both AP and SFO were ablated, while lesion of either of these circumventricular organs in isolation did not influence the feeding responses to PYY3-36-HSA. The PYY3-36-HSA-induced anorectic effect was also reduced during the 3- to 6-h period following subdiaphragmatic vagotomy. Lesions of AP, SFO and AP/SFO as well as subdiaphragmatic vagotomy blunted PYY3-36-HSA-induced expression of c-fos mRNA in specific brain structures including the bed nucleus of stria terminalis, central amygdala, lateral–external parabrachial nucleus and medial nucleus of the solitary tract. In addition, subdiaphragmatic vagotomy inhibited the neuronal activation induced by PYY3-36-HSA in AP and SFO. These findings suggest that the anorectic effect and brain neuronal activation induced by PYY3-36-HSA are dependent on integrity of AP, SFO and subdiaphragmatic vagus nerve.