Luu Kien C, Nie Gui Ying, Hampton Anne, Fu Guo-Qiang, Liu Yi-Xun, Salamonsen Lois A
Prince Henry's Institute of Medical Research, Clayton, Victoria 3168, Australia.
Reproduction. 2004 Oct;128(4):433-41. doi: 10.1530/rep.1.00226.
The endometrium is hostile to embryo implantation except during the 'window of receptivity'. A change in endometrial gene expression is required for the development of receptivity. Calbindin-d9k (CaBP-d9k) and calbindin-d28k (CaBP-d28k) are proteins possessing EF-hand motifs which have high affinity for Ca2+ ions. Previously, it has been demonstrated that, in mouse endometrium, the expression of both calbindins is highly regulated during implantation and that both proteins play critical but functionally redundant roles at implantation. This study was the first to determine the expression of these two calbindins in the human and rhesus monkey endometrium. Initial RT-PCR analysis demonstrated that CaBP-d28k but not CaBP-d9k mRNA expression is detectable in the endometrium of both species. Western blot analysis confirmed the presence of immuno-reactive CaBP-d28k protein in the primate endometrium. Furthermore, the endometrial expression pattern of CaBP-d28k mRNA and protein was examined by Northern blot analysis and immunohistochemistry respectively in both species across the menstrual cycle and during early pregnancy. Semi-quantitative statistical analysis of the immunohistochemistry results revealed that, in the human, CaBP-d28k protein expression was maximal in luminal and glandular epithelium during the mid-secretory phase, coinciding with the time when the endometrium is receptive to embryo implantation. Expression in rhesus monkey showed a similar trend. These results suggest that, in the primate endometrium, only CaBP-d28k is expressed and that the specific regulation of this calbindin is potentially important for the establishment of uterine receptivity.
除了在“着床窗口期”,子宫内膜对胚胎着床具有抗性。子宫内膜接受性的发展需要基因表达发生变化。钙结合蛋白-d9k(CaBP-d9k)和钙结合蛋白-d28k(CaBP-d28k)是具有EF-手基序的蛋白质,它们对钙离子具有高亲和力。此前已证明,在小鼠子宫内膜中,两种钙结合蛋白的表达在着床期间受到高度调控,且这两种蛋白质在着床过程中发挥关键但功能冗余的作用。本研究首次确定了这两种钙结合蛋白在人类和恒河猴子宫内膜中的表达情况。最初的逆转录聚合酶链反应(RT-PCR)分析表明,在这两个物种的子宫内膜中均可检测到CaBP-d28k的mRNA表达,而未检测到CaBP-d9k的mRNA表达。蛋白质印迹分析证实了在灵长类动物子宫内膜中存在免疫反应性CaBP-d28k蛋白。此外,分别通过Northern印迹分析和免疫组织化学方法,在两个物种的整个月经周期和妊娠早期检测了CaBP-d28k mRNA和蛋白的子宫内膜表达模式。免疫组织化学结果的半定量统计分析显示,在人类中,CaBP-d28k蛋白表达在分泌中期的腔上皮和腺上皮中最高,这与子宫内膜接受胚胎着床的时间一致。恒河猴的表达也呈现类似趋势。这些结果表明,在灵长类动物子宫内膜中,仅表达CaBP-d28k,且这种钙结合蛋白的特异性调控可能对子宫接受性的建立具有重要意义。