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Objective: Neuropeptide Y (NPY), given centrally augments food intake and the motivation to work for palatable food. Here, the brain regions were identified through which NPY increases food intake and motivation. Methods: NPY was infused into three brain regions implicated in food intake and motivation: the lateral hypothalamus (LH), nucleus accumbens shell (NAc), and ventral tegmental area (VTA). Motivation for sucrose was assessed using a progressive-ratio schedule of reinforcement in which the effort to obtain successive rewards increased incrementally. To disentangle the effects of NPY on motivation for palatable food from food consumption, free-feeding experiments were performed in which animals had ad libitum access to sucrose pellets. Results: Infusion of NPY into either VTA or NAc increased the motivation to respond for sucrose, whereas infusion of NPY in either NAc or LH increased sucrose consumption. In addition, the effect of intra-VTA NPY on motivation for food was attenuated after pretreatment with the dopamine receptor antagonist alpha-flupenthixol. Conclusions: Specific limbic substrates through which NPY influences consumption of and motivation for palatable food were identified by these data. The motivational effects of NPY are exerted through the VTA, its consummatory effects through the LH, and the NAc is involved in both.