Department of Obstetrics and Gynaecology, University of Otago, Christchurch 8011, New Zealand.
Hum Reprod. 2012 Sep;27(9):2747-55. doi: 10.1093/humrep/des233. Epub 2012 Jun 27.
To use contemporary biochemical markers to characterize mRNA/gene expression in the potentially fertile secretory endometrium to confirm its identification based on histological characteristics in order to develop a clinically applicable test.
Nine, fertile, cycling Caucasian women were sampled from one IVF clinic. Endometrial samples were collected from them in two to four menstrual cycles at 2 and 7 days post first significant rise in blood LH. Separate endometrial glands and stroma populations were obtained by laser microdissection. Linear polymerase chain reaction amplified mRNAs which were hybridized to both Affymetrix U133 Plus2 and Agilent 4 × 44K microarrays followed by gene set analysis. Four histopathologists reviewed the sample set using the same histological criteria to date and characterize the non-receptive and potentially receptive samples.
mRNA expression of microdissected glands and stroma provided molecular signatures that characterized the two specific phases of the cycle with distinct clustering patterns. Cell proliferation and five other associated biological pathways were significantly down-regulated when the endometrium is considered potentially receptive accompanied by an increase in secreted glycoproteins mRNAs in the potentially receptive glands. Reported histological findings identified the presence of one histological feature characteristic of each phase: glandular mitoses indicated a non-receptive endometrium, whereas a potentially receptive endometrium was distinguished by supranuclear vacuolation.
This study defined a transcriptome characteristic of active cell proliferation in the non-receptive samples with a marked overall down-regulation of this pathway in potentially receptive samples-suggesting a transitional state associated with receptivity but not implantation. However, microarrays involve expensive, specialized testing and require significant post-data analysis. Sampling according to endocrinological and molecular prediction improved the consistency of histological assessment and allowed reliable histological markers of glandular mitosis in the non-receptive phase and supranuclear vacuolation of the potentially receptive endometrium to be identified. Thus, histology can provide an affordable, clinically applicable test in the context of reproduction.
利用当代生化标志物来描述有生育能力的分泌期子宫内膜中的 mRNA/基因表达,以确认基于组织学特征的鉴定,并开发一种临床适用的检测方法。
从一家试管婴儿诊所招募了 9 名生育能力正常的白人女性。在血液 LH 首次显著升高后 2 天和 7 天,从她们的 2 至 4 个月经周期中采集子宫内膜样本。通过激光显微切割获得分离的子宫内膜腺体和基质群体。线性聚合酶链反应扩增的 mRNA 与 Affymetrix U133 Plus2 和 Agilent 4 × 44K 微阵列杂交,然后进行基因集分析。四位组织病理学家使用相同的组织学标准回顾样本集,对非接受性和潜在接受性样本进行评估和特征描述。
显微切割腺体和基质的 mRNA 表达提供了分子特征,这些特征可区分周期的两个特定阶段,聚类模式明显不同。当子宫内膜被认为具有潜在接受性时,细胞增殖和其他五个相关的生物学途径显著下调,同时潜在接受性腺体中分泌糖蛋白 mRNA 增加。报告的组织学发现确定了每个阶段存在一个组织学特征:腺体有丝分裂表明非接受性子宫内膜,而潜在接受性子宫内膜的特征是核上空泡。
本研究定义了非接受性样本中活跃细胞增殖的转录组特征,潜在接受性样本中该途径明显下调,表明与接受性而非着床相关的过渡状态。然而,微阵列涉及昂贵的、专业化的测试,并且需要大量的数据分析。根据内分泌学和分子预测进行采样提高了组织学评估的一致性,并允许识别非接受期腺体有丝分裂和潜在接受期子宫内膜核上空泡的可靠组织学标志物。因此,组织学可以在生殖背景下提供一种经济实惠、临床适用的检测方法。