The current study was designed to evaluate the effects of central administration of L-arginine (The precursor of nitric oxide), N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide (NO) synthase inhibitor, selective opioid receptor agonists and involvement of central nitrergic/opioidergic systems on feeding behavior in neonatal layer-type chicks. The results of this study showed that the intracerebroventricular (ICV) injection of L-arginine (400 and 800 nmol) significantly decreased food intake (P < 0.001) but the injection of 200 nmol L-arginine had no effect on cumulative food intake in FD3 chickens (P > 0.05). The ICV injection of L-NAME (200 and 400 nmol) increased food intake (P < 0.001) but 100 nmol of L-NAME had no significant effect (P > 0.05). On the other hand, the co-injection of 100 nmol L-NAME significantly attenuated the anorexigenic effect of 800 nmol L-arginine (P < 0.001). Moreover, the food intake of chicks was significantly decreased by ICV injection of DAMGO (μ-opioid receptor agonist, 125 pmol) (P < 0.001) while both DPDPE (δ-opioid receptor agonist, 40 pmol) and U-50488H (κ-opioid receptor agonist, 30 nmol) significantly stimulated food intake (P < 0.001). In addition, the hypophagic effect of DAMGO was significantly amplified by administration of L-arginine (P < 0.001) while the administration of L-NAME attenuated the hypophagic effect of DAMGO (P < 0.001). In contrast, co-injection of L-arginine or L-NAME with DPDPE had no effect on the hyperphagia induced by DPDPE as well as the hyperphagic effect of U-50488H on food intake was not affected by concurrent injection of L-arginine or L-NAME (P > 0.05). These results suggest that nitrergic and opioidergic systems have an important role on feeding behavior in the CNS of neonatal layer-type chicks and it seems that interaction between them is mediated by μ-opioid receptor.