Stachecki J J, Yelian F D, Leach R E, Armant D R
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, MI 48201.
J Reprod Fertil. 1994 Aug;101(3):611-7. doi: 10.1530/jrf.0.1010611.
This study focused on the effects of ethanol on blastocyst outgrowth and implantation in mice. Blastocysts were exposed to ethanol in Ham's F10 medium and then cultured free of ethanol on fibronectin-coated Petri dishes to assess trophoblast cell adhesion and migration. The time necessary for half of the embryos to outgrow was significantly less (P < 0.05) following treatment with 0.1%, 0.2%, 0.4% or 1.0% (w/v) ethanol for either 5 min or 24 h compared with controls. The rate of trophoblast cell migration was determined by measuring the mean area of outgrowing embryos using an image analysis system. Blastocysts exposed to ethanol for 5 min produced a greater (P < 0.05) average outgrowth area than did stage-matched controls. Acceleration of blastocyst cavitation by ethanol is known to be associated with an increase in the intracellular concentration of calcium. Here, treatment with the calcium ionophore A23187 stimulated (P < 0.05) trophoblast outgrowth and accelerated (P < 0.05) the rate of cell migration. In an attempt to correlate the effect of ethanol on outgrowth in vitro with implantation in utero, cultured blastocysts were either not exposed to ethanol or exposed to 0.1% ethanol for 5 min and transferred 24 h later to the uteri of pseudopregnant dams. The implantation rate (39.4%, n = 376) and the rate of development to term (2.45 pups per mouse, n = 20) were higher (P < 0.05) in mice receiving ethanol-treated embryos compared with those receiving control embryos (20.8%, n = 331; 1.16 pups per mouse, n = 18, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)
本研究聚焦于乙醇对小鼠囊胚孵出及着床的影响。将囊胚置于哈姆氏F10培养基中用乙醇处理,然后在包被纤连蛋白的培养皿中无乙醇培养,以评估滋养层细胞的黏附和迁移。与对照组相比,用0.1%、0.2%、0.4%或1.0%(w/v)乙醇处理5分钟或24小时后,半数胚胎孵出所需时间显著缩短(P<0.05)。用图像分析系统测量孵出胚胎的平均面积来确定滋养层细胞迁移率。暴露于乙醇5分钟的囊胚产生的平均孵出面积比同期对照更大(P<0.05)。已知乙醇加速囊胚空化与细胞内钙浓度增加有关。在此,用钙离子载体A23187处理可刺激(P<0.05)滋养层孵出并加速(P<0.05)细胞迁移率。为了将乙醇对体外孵出的影响与子宫内着床相关联,将培养的囊胚要么不暴露于乙醇,要么暴露于0.1%乙醇5分钟,24小时后移植到假孕母鼠子宫内。与接受对照胚胎的小鼠相比(着床率分别为20.8%,n = 331;每只小鼠产仔1.16只,n = 18),接受乙醇处理胚胎的小鼠着床率(39.4%,n = 376)和足月发育率(每只小鼠产仔2.45只,n = 20)更高(P<0.05)。(摘要截断于250字)