In the present investigation, in vivo microdialysis was used to measure the concentration of estrogen in the parabrachial nucleus (PBN) and plasma of male and ovariectomized female Sprague-Dawley rats supplemented with either estrogen (OVX-E(2)) or saline (OVX-S) following visceral afferent activation. Analysis of dialysate samples prior to vagal stimulation and in non-stimulated controls revealed a continuous concentration of estrogen in the PBN for all treatment groups (male, 38 +/-4 pg ml(-1); OVX-E(2), 38+/-5 pg ml(-1); OVX-S, 33 +/- 4 pg ml(-1)). This concentration of estrogen in the PBN was significantly increased during vagal stimulation in all groups (male, 64+/-4 pg ml(-1); OVX-E(2), 104+/-9 pg ml(-1); OVX-S, 80+/-6 pg ml(-1); P<0.05) and returned to pre-stimulation values within 2 h following termination of the stimulation. Immunohistochemical analysis revealed that estrogen receptor (ERalpha and ERbeta) density in males and ovariectomized saline-replaced female rats was significantly lower than that of estrogen-replaced female rats. These results suggest that estrogen is released into the PBN by an increase in visceral afferent traffic, however, alterations in estrogen receptor populations in the PBN may contribute to an attenuated physiological role of estrogen in the PBN of male and saline-replaced ovariectomized female rats.