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Chunk #0 — RESEARCH DESIGN AND METHODS — Animals.

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Nicotine induces negative energy balance through hypothalamic AMP-activated protein kinase.
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We used adult male Sprague-Dawley rats (9–11 weeks old; Animalario-General USC). Rats were housed in a temperature-controlled (22°C) room, with a 12-h light/dark cycle (7:00 to 19:00). The experiments were performed in agreement with the International Law on Animal Experimentation and approved by the USC Bioethics Committee and the Ministry of Science and Innovation of Spain (PS09/01880). For all of the experimental groups and analytic methods, we used 9–16 rats per group.