To study a potential locus of action of opioids in the rat dentate gyrus, we examined the localization of the delta opioid receptor (DOR) by immunocytochemistry. Two antisera raised to unique, non-overlapping peptide sequences located within the extracellular N-terminal sequence of DOR were tested. By light microscopy, numerous neurons in the central hilar region were intensely labeled for DOR, while the granule cell layer contained light DOR immunoreactivity. To further characterize hilar neuron cell types which contained DOR, sections through the dentate gyrus were double labeled using immunofluorescence with antisera to DOR and either gamma-aminobutyric acid (GABA), neuropeptide Y (NPY), or somatostatin-28 antisera. Most DOR-labeled perikarya also contained GABA and NPY, while a subpopulation contained somatostatin. Electron microscopic examination of sections labeled for DOR revealed that the immunoreactivity was common in profiles which exhibited the morphological characteristics of granule cells, as well as those of non-granule cells. DOR immunoreactivity was located at postsynaptic sites within neuronal perikarya (2%), dendrites (27%), and dendritic spines (22%); as well as in presynaptic axon terminals (25%) and glia (23%) (n = 279). In dendrites and dendritic spines, DOR immunoreactivity was most often associated with the plasmalemmal surface near asymmetric synapses. In axon terminals, DOR immunoreactivity primarily surrounded small, clear vesicles, and was less consistently found on the plasmalemmal surface. The distribution of DOR-labeled profiles overlapped with, but was not restricted to regions known to contain enkephalin. These data suggest that opiates acting at the DOR can modulate both hilar neurons and granule cells both pre- and postsynaptically.