Evidence from animal studies suggests that opioidergic system and gherlin have a regulatory role in food intake, but their interaction(s) have not been studied in laying chickens. So in this study, 4 experiments (each included 4 groups) were designed. The first experiment was performed to evaluate the effect of ghrelin on the cumulative food intake. Experiments 2 to 4 were designed to investigate the possibility of μ, δ, or κ opioid receptors mediating ghrelin-induced hypophagia. All drugs were injected intracerebroventricularly (ICV) at 5 days of age. The results of this study showed that the ICV injection of 1.5 nmol ghrelin did not affect cumulative food intake. But ICV injection of ghrelin with doses of 3 and 6 nmol significantly reduced the cumulative food intake (p<0.05). However, co-injection of ghrelin with NTI, and nor-BNI did not show a significant change in decreased food intake compared to ghrelin. Also, opioid μ receptor gene expression significantly increased (p<0.05), but δ and κ opioid receptors gene expression did not significantly change. These results indicated the opioidergic system is involved in developing ghrelin-induced hypophagic effects in laying chickens. Accordingly, this effect of ghrelin to modify the nutritional behavior is possibly mediated by opioid μ receptor.