Oxytocin receptors are regulated during parturition and lactation. Gonadal steroids are thought to be key players in this regulation. It is not well documented how oxytocin receptor gene expression is regulated in the CNS. In this study we analyzed potential estrogen effects on the oxytocin receptor mRNA levels in some areas integral to the limbic-hypothalamic system, namely the ventromedial nucleus of the hypothalamus (VMH), posterior medial nucleus of amygdala (MeAmyg), and arcuate nucleus (ARC), as well as the caudate putamen (CPu), CA1 region of the hippocampus, anterior pituitary, and uterine tissue of ovariectomized (OVX) female rats. By in situ hybridization we observed a 4.4-fold increase in oxytocin receptor mRNA levels in the VMH after 48 h of estrogen treatment when compared to OVX rats. Smaller increases were observed in the MeAmyg, hippocampus, and anterior pituitary (3.18, 1.76, and 2.55, respectively). No changes in oxytocin receptor mRNA levels were observed in the CPu or ARC after estrogen treatment. A similar finding resulted from slot-blot analysis of total mRNA extracts. In uterine tissue, 48 h of estrogen treatment increased oxytocin receptor mRNA level in the myometrium (3.13-fold). No changes in oxytocin receptor mRNA levels were observed after 12 and 24 h of estrogen treatment. These findings suggest that the estrogenic regulation of oxytocin receptor binding in both CNS and uterine tissues may in part be mediated by de novo synthesis of oxytocin receptor mRNA or by alterations in the stability of oxytocin receptor gene transcripts.