A striking sexual dimorphism exists in the distribution of Met-enkephalin (m-ENK) immunoreactive fibers in the preoptic area of the rat brain. A dense plexus of m-ENK fibers, approximately 100 microns wide, is present in the periventricular part of the preoptic area (pePOA) in adult females, but not in males. In the present study, we have examined the time of first expression of this female-typical system during the course of normal brain development. The female-typical plexus of m-ENK fibers in the pePOA is not expressed prepubertally, but first becomes evident during the late peri-pubertal period (usually by 40 days of age). In adult females, the maintenance of immunohistochemically detectable levels of m-ENK in this fiber system is dependent upon the presence of gonadal steroids, especially estradiol. Therefore, we examined whether exposure of pre-pubertal females to estradiol would result in precocious expression of the m-ENK fiber plexus. The results of this experiment demonstrate that exposure to estradiol for 7 days induces the full expression of the m-ENK fiber system in the pePOA of prepubertal females, such that it was indistinguishable from that seen in mature animals. These results demonstrate that while the sexually dimorphic m-ENK system of the pePOA is not normally expressed prepubertally, the neural substrate is nevertheless in place and capable of being activated by exposure to exogenous estradiol.