The periventricular preoptic area (pePOA) is a sexually dimorphic component of the rat forebrain that contains a sexually dimorphic Met-enkephalin immunoreactive (ENK-ir) fiber plexus. This plexus is especially dense in the female while only scattered ENK-ir fibers are present in the pePOA of the male. Abundant estrogen receptive neurons are located in the pePOA of both the female and male. This experiment was conducted to determine if estrogen receptive neurons in the pePOA are postsynaptic targets of ENK-ir terminals. Double label ultrastructural localization of estrogen receptor (ER)-ir neurons and ENK-ir fibers was performed using the chromogens 3,3',5,5'-tetramethylbenzidine (TMB) and diaminobenzidine tetrahydrochloride (DAB), respectively. TMB-stained ER-ir neurons contained electron dense crystalline spicules located predominantly in their nuclei. Flocculent DAB reaction product was distributed over membraneous structures in ENK-ir fibers and terminals. Numerous ER-ir neurons were present in the pePOA of the male and female. In females, many ENK-ir terminals, both synaptic and non-synaptic, contacted the perikarya of ER-ir neurons. In contrast, many fewer ENK-ir terminals made contact on ER-ir neurons in the male. Thus, these results provide morphological evidence that ENK-ir neurons can regulate ER-ir neurons in the pePOA. Moreover, because expression of the ENK-ir pePOA fiber plexus is estrogen-sensitive in the female, these results suggest strongly that estrogen may regulate these neurons both pre- and postsynaptically. Finally, these results provide additional evidence for the involvement of the sexually dimorphic pePOA ENK-ir fibers plexus in the control of estrogen-mediated function in the female.