Using the technique of in vitro steroid autoradiography, the localization and modulation of nuclear estrogen binding sites has been studied in normal human cervix and vagina during the menstrual cycle, pregnancy, and the menopause. Marked differences occur in nuclear estrogen binding between these two organs. Nuclear estrogen binding varies throughout the menstrual cycle in the vaginal epithelium, whereas vaginal stromal cells consistently exhibit nuclear estrogen binding throughout the cycle. In contrast, the cervical squamous and columnar epithelia show much less cyclic variability in nuclear estrogen binding sites. As in the vagina, the cervical stroma consistently binds estrogen. High levels of nuclear estrogen binding sites are found in the vagina of the postmenopausal patient, and lower levels of binding occur postpartum. The implications of these localizations, with special reference to the role of the cervical and vaginal stroma, are discussed.