Estrogen has been implicated in brain function related to stress responses. We investigated whether estrogen affects psychological stress-induced activation of peptide-containing or nitric oxide-producing neurons in the paraventricular hypothalamic nucleus (PVN) in ovariectomized (OVX) rats, both placebo-treated (OVX + Pla) and estrogen-treated (OVX + E2) by comparison of c-Fos expression using immunohistochemistry. Cage-switch stress increased activation in oxytocinergic neurons in the parvocellular PVN (pPVN) in OVX + Pla, but not in that of OVX + E2, rats. Moreover, the stress-induced activation in NADPH-diaphorase-positive neurons in the pPVN was larger in the OVX + E2 than in the OVX + Pla group. These findings suggest that estrogen attenuates the activation of oxytocinergic neurons in the pPVN, at least in part via nitric oxide.