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Peptidergic systems in the medial hypothalamus (MH) are involved in feeding and energy homeostasis. High protein diets have been suggested to aid in weight loss by increasing satiety and decreasing food intake in human studies as well as animal models. The objective of this study was to evaluate the pattern of changes in orexigenic and anorexigenic neuropeptides (NPs) in the MH of rats fed a high protein diet (HP; 30% protein: 38% CHO: 32% fat) versus a standard protein diet (SP; 15:59:26). Our study focused on the pattern of NP change in the MH when the high protein diet was fed. After 3 weeks rats were sacrificed after an overnight fast, the animals were placed into 3 groups: immediately sacrificed, 1hr or 3hr following a 3g meal. At sacrifice the MH was removed and homogenized in acetic acid. Following centrifugation, proteins were extracted and analyzed for Neuropeptide Y (NPY), Orexin A (OxA; also known as Hypocretin-1), Cocaine-Amphetamine Related Transcript (CART), and α-Melanocortin Stimulating Hormone (α-MSH) by radioimmunoassay. Total food intake levels during the last week were ~10% (p=0.01) lower in the HP group. NPY was 20% lower and α-MSH was 12% higher after 3 weeks of HP. This relationship also remained at 1 hour and to a lesser extent 3 hours post-meal. Our results indicate that the expression of the orexigenic NPs (NPY and OxA) was decreased in rats chronically fed the HP diet, while the expression of anorexigenic NPs (CART and α-MSH) was elevated in these same animals. The comprehensive pattern of changes in the appetite-related NPs might contribute to the reduced food intake in animals fed a HP diet. NIH DK59755