Gruidl M, Buyuksal A, Babaknia A, Fazleabas A T, Sivarajah S, Satyaswaroop P G, Tabibzadeh S
Department of Pathology, Moffitt Cancer Center, Tampa, FL 33612, USA.
Mol Hum Reprod. 1997 Apr;3(4):333-42. doi: 10.1093/molehr/3.4.333.
Human endometrium undergoes sequential changes during the menstrual cycle and becomes receptive to implantation during a defined period in the secretory phase. We attempted to identify the genes expressed during this period by representational difference analysis (RDA). When the cDNAs of a proliferative endometrium were used as the driver and the cDNAs of a post-ovulatory day 5 endometrium were used as the tester, a number of bands were identified by RDA. DNA of the cloned RDA products revealed that the majority of the clones contained a fragment of a cDNA identical to that of a crystallin B chain. Northern blot analysis showed that the expression of the alpha crystallin B chain mRNA was absent during the proliferative phase. The expression of the mRNA of alpha crystallin B chain first appeared in the secretory phase, progressively increased during this phase and peaked in the late secretory endometria. The pattern of expression of alpha crystallin B chain mRNA in the endometrium of mature cycling baboons (Papio anubis) was similar to that seen in human endometrium. As revealed by Western blot analysis, the expression of the alpha crystallin B chain protein in human endometrium followed a pattern of expression similar to its mRNA. At the cellular level, the immunoreactive protein first appeared in the surface epithelial cells of human endometrium within the implantation window without significant immunoreactivity in the underlying glandular cells. During the mid- and late secretory phases, the intensity of staining in the epithelial cells was enhanced and an intense immunoreactivity was developed in the glandular epithelium, alpha crystallin B chain was virtually an epithelial product and no immunoreactivity for this protein was detectable in the stromal cells, endothelial cells or lymphoid cells. The expression of alpha crystallin B chain could be regulated, by medroxy progesterone acetate as well as by oestrogen withdrawal, in human endometrial carcinoma cells (EnCa-101), transplanted to nude mice. Based on the data presented here, the known function of alpha crystallin B chain and its distinct pattern of expression in human endometrium, we suggest that this protein is an important factor within the molecular repertoire that makes endometrium receptive to implantation.
人类子宫内膜在月经周期中会经历一系列变化,并在分泌期的特定时间段变得易于着床。我们试图通过代表性差异分析(RDA)来鉴定在此期间表达的基因。当将增殖期子宫内膜的cDNA用作驱动子,排卵后第5天子宫内膜的cDNA用作测试子时,通过RDA鉴定出了许多条带。克隆的RDA产物的DNA显示,大多数克隆包含与晶状体蛋白B链cDNA相同的片段。Northern印迹分析表明,增殖期不存在α晶状体蛋白B链mRNA的表达。α晶状体蛋白B链mRNA的表达首先出现在分泌期,在此期间逐渐增加,并在分泌晚期子宫内膜中达到峰值。成熟的周期性狒狒(Papio anubis)子宫内膜中α晶状体蛋白B链mRNA的表达模式与人类子宫内膜中所见相似。Western印迹分析显示,人类子宫内膜中α晶状体蛋白B链蛋白的表达遵循与其mRNA相似的表达模式。在细胞水平上,免疫反应性蛋白首先出现在着床窗内人类子宫内膜的表面上皮细胞中,而在下方的腺细胞中无明显免疫反应性。在分泌中期和晚期,上皮细胞中的染色强度增强,腺上皮中出现强烈的免疫反应性,α晶状体蛋白B链实际上是一种上皮产物,在基质细胞、内皮细胞或淋巴细胞中未检测到该蛋白的免疫反应性。在移植到裸鼠体内的人子宫内膜癌细胞(EnCa-101)中,α晶状体蛋白B链的表达可受醋酸甲羟孕酮以及雌激素撤退的调节。基于此处提供的数据、α晶状体蛋白B链的已知功能及其在人类子宫内膜中独特的表达模式,我们认为该蛋白是使子宫内膜易于着床的分子库中的一个重要因素。