In the literature, neuropeptide Y (NPY) has been described in the brain and peripheral nerves. More recently, it has also been detected in endocrine cells of hamster, embryonic mouse, and rat pancreas. However, the presence of NPY in avian embryos and the possible colocalization of this peptide with the other pancreatic hormones have not been reported previously. In this study, NPY presence was studied by immunocytochemical methods in the endocrine pancreas of domestic duck during pre- and postnatal development. NPY immunoreactivity (IR) was detected in embryos and adult animals. Around hatching the intensity of IR in endocrine cells decreased. Double immunohistochemical staining revealed that: 1) NPY-IR is extensively colocalized in small and mixed islets with insulin-IR both in embryos and in adults; and 2) in early embryos NPY-IR occasionally colocalized with glucagon and somatostatin. In early embryos, the colocalization of NPY-IR with several pancreatic hormones could be related to the presence of multi-hormonal progenitor cells. The close relation between insulin and NPY, both in embryos and adults, led us to hypothesize a key role for NPY on insulin cells of duck pancreas.