Serial analysis of gene expression in mouse uterus at the implantation site
Serial analysis of gene expression in mouse uterus at the implantation site
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小鼠子宫植入部位基因表达的系列分析
DOI：
10.1074/jbc.m511512200
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发表时间：
2006-04-07
影响因子：
4.8
通讯作者：
Yang, ZM
中科院分区：
文献类型：
--
作者：
Ma, XH;Hu, SJ;Yang, ZM
虽然已有寡核苷酸芯片、基因芯片、差异显示反转录-聚合酶链式反应等方法被用于筛选着床相关分子，但胚胎着床发生的机制仍不清楚。本研究的目的是通过基因表达序列分析(SAGE)来分析妊娠第5天小鼠子宫着床点和着床点基因表达的差异。在我们的两个11bp标签的SAGE文库中，植入部位的标签总数为48,121个，植入部位的标签总数为50,227个。有1039个标签在植入部位特异表达，1252个标签在植入部位特异表达。根据p值，有195个标签在植入部位显著上调，261个标签在植入部位显著上调，其中100个基因在植入部位单一匹配，127个基因在植入部位单一匹配。通过逆转录-聚合酶链式反应，在14个有显著变化的基因上验证了植入部位和植入间部位的标签比例。原位杂交结果显示，1810014L12Rik、Psmb5、CD63、NPM1、Fads3和Tagln2基因在植入部位的表达高于植入间部位。与植入部位相比，Ddx39在妊娠第5天植入部位的瘤下基质细胞有较强的表达。Ddx39在着床部位的表达是由活跃的囊胚特异性诱导的。此外，雌激素显著上调去卵巢小鼠Ddx39的表达。在我们的SAGE数据中，在小鼠子宫中发现了许多与着床相关的基因。我们的数据可能为未来胚胎着床的研究提供有价值的来源。
Although oligonucleotide chips, cDNA microarrays, differential display reverse transcription-PCR, and other approaches have been used to screen implantation-related molecules, the mechanism by which embryo implantation occurs is still unknown. The aim of this study was to profile the differential gene expression between interimplantation site and implantation site in mouse uterus on day 5 of pregnancy by serial analysis of gene expression (SAGE). In our two SAGE libraries of 11-bp tags, the total numbers of tags sequenced were 48,121 for the interimplantation site and 50,227 for the implantation site. There were 1,039 tags specifically expressed at interimplantation site, and 1,252 tags specifically expressed at the implantation site. Based on the p value, there were 195 tags significantly up-regulated at the interimplantation site and 261 tags significantly up-regulated at the implantation site, of which 100 genes were single matched at the interimplantation site and 127 genes were single matched at the implantation site, respectively. By reverse transcription-PCR, the tag ratio between the implantation site and interimplantation site was verified on 14 significantly changed genes. Using in situ hybridization, 1810014L12Rik, Psmb5, Cd63, Npm1, Fads3, and Tagln2 were shown to be highly expressed at the implantation site compared with the interimplantation site. Compared with the interimplantation site, Ddx39 was strongly expressed in the subluminal stromal cells at the implantation site on day 5 of pregnancy. Ddx39 expression at the implantation site was specifically induced by active blastocysts. Additionally, Ddx39 expression was significantly up-regulated by estrogen in the ovariectomized mice. In our SAGE data, many implantation-related genes were identified in mouse uterus. Our data could be a valuable source for future study on embryo implantation.