Prenatal development of rat pancreatic endocrine cells was investigated by the immunoperoxidase technique and the following results were obtained: 1) Glucagon immunoreactive cells are first endocrine element of the pancreas appearing already on day 11 of gestation, when the dorsal pancreatic bud is still quite small and the ventral pancreatic primordium is hardly swollen out. Most of the glucagon reactive cells are located in the epithelium of the foregut and the dorsal pancreatic bud but a few of them are attached to the base of the epithelium from the outside. 2) Insulin and pancreatic polypeptide (PP) immunoreactive cells are first demonstrable in small cell clusters budding from the exocrine tubules on day 14, whereas somatostatin and gastrin reactive cells on day 17 and 18, respectively. These findings support the hypothesis that the majority of pancreatic endocrine cells is derived from the epithelium of the foregut. 3) PP reactive cells, appearing first on day 14, assume gradually a peripheral position in the growing islet of Langerhans. Immediately before birth they attain the population and flattened cell shape comparable to those in adult pancreas. Their counterpart in the duodenum is found as open type basal-granulated cells in the rat fetus. 4) Noteworthily, glucagon-like immunoreactivity is found in some neuron-like cells of the Auerbach's plexus between the muscle layers of the duodenum on day 19.