Kowalczyk C L, Stachecki J J, Schultz J F, Leach R E, Armant D R
C. S. Mott Center for Human Growth and Development, Department of Obstetrics and Gynecology, Wayne State University, Detroit, Michigan 48201, USA.
Alcohol Clin Exp Res. 1996 May;20(3):566-71. doi: 10.1111/j.1530-0277.1996.tb01094.x.
During in vitro culture of murine preimplantation embryos, we have observed that exposure to 0.1% ethanol induces an immediate increase in intracellular calcium levels and subsequently accelerates embryogenesis. If the observed effects of ethanol on developing embryos is mediated by its membrane disordering potency, we hypothesized that the relative membrane disordering potencies of related alcohols would correspondingly effect embryonic intracellular calcium levels and developmental rates. Two-cell embryos were exposed to 0.1% ethanol or 0.05 to 1.0% (w/v) n-butanol, n-propanol, isopropanol, 1,2-propanediol, glycerol, or methanol for 24 hr at 37 degrees C, and development to the blastocyst stage was monitored after 5 days. n-Butanol, n-propanol, isopropanol, and methanol treatment caused a dose-dependent inhibition (p < 0.01) of development to the blastocyst stage, whereas 1,2-propanediol or glycerol neither accelerated nor inhibited development. In a second experiment, 8-cell morulae were treated with 1,2-propanediol or glycerol, and cavitation rates were examined. There was no significant difference from control embryos in the onset of cavitation or the blastocoel expansion rate of 1,2-propanediol- or glycerol-exposed embryos, whereas exposure to 0.1% ethanol accelerate cavitation (p > 0.05). In a third experiment, morulae were exposed to 0.1% or 1.0% of each alcohol and were monitored for changes in intracellular calcium levels using the fluorescent indicator, fluo-3-acetoxymethyl ester. There was an immediate increase in intracellular calcium levels when morulae were treated with 1.0% ethanol or n-butanol, but only ethanol induced an increase (p < 0.05) in the level of intracellular calcium at 0.1%. These data suggest that ethanol is unique in its ability to accelerate embryogenesis and that the membrane disordering potency of ethanol does not directly underlie its effects on intracellular calcium release and the acceleration of preimplantation development.
在小鼠植入前胚胎的体外培养过程中，我们观察到，暴露于0.1%的乙醇会导致细胞内钙水平立即升高，随后加速胚胎发育。如果观察到的乙醇对发育中胚胎的影响是由其膜紊乱能力介导的，我们推测相关醇类的相对膜紊乱能力会相应地影响胚胎细胞内钙水平和发育速率。将二细胞胚胎在37℃下暴露于0.1%的乙醇或0.05%至1.0%（w/v）的正丁醇、正丙醇、异丙醇、1,2 - 丙二醇、甘油或甲醇中24小时，5天后监测发育至囊胚阶段的情况。正丁醇、正丙醇、异丙醇和甲醇处理导致发育至囊胚阶段呈剂量依赖性抑制（p < 0.01），而1,2 - 丙二醇或甘油既不加速也不抑制发育。在第二个实验中，用1,2 - 丙二醇或甘油处理8细胞桑椹胚，并检查空化率。暴露于1,2 - 丙二醇或甘油的胚胎在空化开始或囊胚腔扩张率方面与对照胚胎没有显著差异，而暴露于0.1%的乙醇会加速空化（p > 0.05）。在第三个实验中，将桑椹胚暴露于每种醇类的0.1%或1.0%中，并使用荧光指示剂fluo - 3 - 乙酰氧基甲酯监测细胞内钙水平的变化。当桑椹胚用1.0%的乙醇或正丁醇处理时，细胞内钙水平立即升高，但只有乙醇在0.1%时会导致细胞内钙水平升高（p < 0.05）。这些数据表明，乙醇在加速胚胎发育的能力方面是独特的，并且乙醇的膜紊乱能力并非其对细胞内钙释放和植入前发育加速作用的直接基础。
