Hoversland R C, Dey S K, Johnson D C
J Reprod Fertil. 1982 Sep;66(1):259-63. doi: 10.1530/jrf.0.0660259.
Rabbit blastocysts were homogenized by sonication, and centrifuged at 105 000 g for 60 min. The pellet was resuspended and incubated in phosphate buffer containing [1 beta-3H]testosterone and a NADPH generating system. The amount of 3H2O produced was determined by liquid scintillation counting. Enzyme activity was calculated, after subtracting blank values obtained with boiled embryos, and expressed as pig testosterone aromatized per embryo per hour. Aromatase activity was undetectable to low on Day 5 and increased on Day 6 of pregnancy. There was a 10-fold increase in activity in Day-6 embryos cultured for 24 h, with a further 6-fold increase in activity in Day-6 embryos cultured for 48 h. The enzyme had an apparent Km of 0.77 microM and was completely inhibited by an aromatase inhibitor. The results clearly indicate that the rabbit blastocyst has an increasing capacity for aromatization of testosterone at about the time of implantation.
兔胚泡经超声处理匀浆，然后在105000g条件下离心60分钟。将沉淀重悬于含有[1β-3H]睾酮和NADPH生成系统的磷酸盐缓冲液中进行孵育。通过液体闪烁计数法测定产生的3H2O量。在减去用煮沸胚胎获得的空白值后计算酶活性，并表示为每个胚胎每小时芳香化的猪睾酮量。妊娠第5天芳香化酶活性检测不到或很低，在妊娠第6天增加。培养24小时的第6天胚胎活性增加了10倍，培养48小时的第6天胚胎活性进一步增加了6倍。该酶的表观Km为0.77微摩尔，被芳香化酶抑制剂完全抑制。结果清楚地表明，兔胚泡在着床时左右对睾酮的芳香化能力不断增强。
