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  • Title: Estrogen stimulation of deoxyribonucleic acid synthesis in uterine epithelial cells which lack estrogen receptors.
    Author: Bigsby RM, Cunha GR.
    Journal: Endocrinology; 1986 Jul; 119(1):390-6. PubMed ID: 3720669.
    Abstract:
    Autoradiographic studies from this laboratory have previously indicated that the uterine epithelium of the neonatal mouse is devoid of estrogen receptors. The neonatal murine uterus is composed of an undifferentiated mesenchyme and a simple columnar epithelium lining the lumen. In the present study, a method for measuring whole cell uptake of 16 alpha-[125I]iodoestradiol ([125I]iodo-E2) was developed and applied to enzymatically separated, isolated epithelial and mesenchymal cells of neonatal (4-5 days postnatal) uteri. Epithelial cells and fibromuscular cells (stromal and myometrial cells) from uteri of 21-day-old animals were used to validate the assay method. A component of the uptake of [125I]iodo-E2 by cells from 21-day-old uteri was shown to be specific, saturable, and of high affinity. Kd values for specific uptake by uterine mesenchymal and epithelial cells were 1.2-1.3 nM. Maximal specific uptake was 9.3 and 4.2 fmol/micrograms DNA for epithelial and mesenchymal cells, respectively. The uterine epithelial cells of 4- and 5-day-old mice showed no measurable specific uptake of [125I]iodo-E2, while the mesenchymal cells from these animals had a maximal specific uptake of 7.9 fmol/micrograms DNA, with a Kd of 1.3 nM. DNA synthesis increased within the uterine epithelium of neonatal mice after estrogen treatment. The thymidine labeling index was doubled 10 h after a single dose of diethylstilbestrol (DES) and returned to pretreatment values by 18 h. The epithelial mitotic index was also 2-fold higher than control values 16-18 h after DES treatment. The increase in the thymidine labeling index was specific to estrogen treatment. DES did not induce the production of estrogen receptors in neonatal uterine epithelium. Epithelial cells of 5-day-old mice that were treated with DES showed no specific [125I]iodo-E2 uptake, while whole cell uptake by mesenchymal cells from these animals exhibited a specific, high affinity component, with a maximal binding of 8.4 fmol/micrograms DNA. Autoradiographic analysis of [3H]estradiol uptake by uterine tissues from 5-day-old mice 12 h after DES treatment did not show nuclear concentration of the steroid in the epithelial cells. These results indicate that the uterine epithelium of the neonatal mouse is indeed devoid of estrogen receptors, and yet the rate of DNA synthesis within this tissue is responsive to estrogen stimulation. The epithelial cells remain devoid of estrogen receptors after DES stimulation, indicating that intraepithelial estrogen receptors are not required for induction of DNA synthesis in these cells in situ. Possible mechanisms by which this phenomenon may occur are discussed.
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