Acute effects of estradiol-17 on plasma volume and uterine cell proliferation in sheep.
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Ovarian steroids play an important role in increasing plasma volume in pregnant females and preparing the uterus for implantation. We hypothesized that a short duration of increased estradiol-17 (E2) would increase plasma volume and uterine cell proliferation in ovariectomized ewes. Adult non-pregnant Romanov ewes (n=15) were ovariectomized. After ovariectomy, ewes were individually housed and were offered water at ad libitum intake and were fed a pelleted diet at maintenance once daily according to body weight. After at least 30 days post-ovariectomy ewes were fasted and received an implant placed in the axillary region that contained 100mg of E2 (E2; n=8) or a sham implant with no E2 (CON, n=7). After 24h, ewes were weighed prior to plasma volume measurement procedures. Plasma volume was determined using the Evans blue dye method. Blood samples were taken at 0 (pre dye injection), 5, 10, 15, 20, 25, 30, 40, 50 and 60min after dye injection. After the final blood collection, ewes were euthanized with an overdose of sodium pentabarbital and uterine weights were recorded. Uterine cross-sections were fixed in formalin for immunohistochemical localization of Ki67 (a marker of proliferating cells) followed by image generation of luminal epithelium and endometrial stroma (5 areas each/tissue section) and analysis to determine the proportion of proliferating cells. Plasma volume tended to be greater in E2 vs CON (2.750.11 vs. 2.540.12L, P=0.07) and uterine weights were greater in E2 vs CON (27.252.35 vs. 17.352.51g, P<0.01). Water intake after implant placement was similar in E2 and CON (3.85 vs. 4.870.67L; P=0.28). Cell proliferation in the luminal epithelium was greater in E2 vs CON (6.55 vs. 1.21.75%, P=0.02) and stromal cells tended to be greater in E2 vs CON (0.59 vs 0.370.06%, P=0.07). Our results demonstrate that E2-treatment tends to increase plasma volume acutely and increases uterine cell proliferation in ewes.
