Obesity was shown to be associated with facilitated gastric emptying, which could either be the cause or the consequence of obesity.1 The anorexigenic neuropeptide, nesfatin-1 (NES-1), when given centrally, suppresses gastric contractions and gastric motility2. High fat diet (HFD) in mice was shown to decrease plasma NES-1 levels.3 However, the impact of HFD on NES-1 induced inhibition of gastric motility, and the peripheral mechanisms underlying the inhibitory effect of NES-1 on gastric emptying were not elucidated yet. In the present study, the involvement of cholecystokinin (CCK)-1, CCK-2 and glucagon-like peptide-1 (GLP-1) receptors in non-caloric liquid emptying and the effect of HFD on NES-1-induced alterations in gastric emptying were investigated. Male Sprague-Dawley rats (10-week-old) were fed with normal diet (ND, 2.7% fat, n=8) or HFD (45% fat, n=8) for 8 weeks. Under ketamine and chlorpromazine (100 and 10 mg/kg,intraperitoneally) anaesthesia, a Gregory cannula was installed in the gastric corpus, and one week later an intracerebroventricular (icv) cannula was placed. After 2 weeks, gastric emptying (GE) rate of saline was determined from the volume and absorbance of fluid recovered 5 min after instillation of phenol red-added saline into the stomach. Gastric emptying was commenced 5 min after icv NES-1 (0.05 µg/0.5 µl) or saline (0.5 µl) injection, which were preceded with subcutaneous injections of either saline or GLP-1 receptor antagonist exendin 9-39 (30 µg/kg) or CCK-1 receptor antagonist devazepide (1 mg/kg) or gastrin/CCK-2 receptor antagonist YM022 (1 mg/kg). At the end of the experiments, verification of icv cannula placement was made using methylene blue. In ND-receiving groups, central NES-1 administration significantly decreased gastric emptying rate (2.62 ± 0.02 ml/5min) compared to icv saline administration (3.06 ± 0.08 ml/5min). None of the antagonists changed the GE rate in icv saline administered rats, but NES-1-induced delay in GE rate was abolished by GLP-1 (3.06 ± 0.13 ml/5 min), CCK-1 (3.25 ± 0.13 ml/5 min, p <0.05) and CCK-2 (3.35 ± 0.13 ml/5 min, p <0.05) receptor antagonist. On the other hand, GE rates in HFD-fed rats were not different among centrally NES-1 (2.95 ± 0.26 ml/5 min) or saline (2.95 ± 0.26 ml/5 min) injected ones, and were similar to saline-injected ND-fed rats, while antagonists did not alter GE rates of HFD-fed rats injected icv with NES-1 or saline. The findings of current study demonstrate that NES-1-induced delay in gastric emptying was mediated via the involvement of GLP-1, CCK-1 and CCK-2 receptors, while this inhibitory effect of NES-1 on gastric emptying rate was not evident when rats were on HFD, suggesting that prolonged consumption of HFD abolishes the inhibitory role of anorexigenic nesfatin-1 on gastric motility and facilitates gastric emptying.