Gene expression profiling of uterine stromal cells isolated from Msx1Msx2 floxed and Msx1Msx2 ablated mice on day 4 of pregnancy

Our preliminary study revealed that the homeobox transcription factors, Msx1 and Msx2, are expressed in the mouse uterus during early pregnancy. Further, conditional deletion of Msx1 and Msx2 in mouse uterus leads to implantation failure due to impaired uterine epithelial receptivity. To identify the downstream targets of Msx1Msx2 in the uterus, we performed gene expression profling of uterine stromal cells isolated from Msx1Msx2-null mice and the corresponding controls on day4 of pregnancy (the time of implantation). The microarray results revealed elevated expression of mRNAs corresponding to several members of the fibroblast growth factor family and Wnts in uterine stroma of Msx1Msx2-ablated mice. We performed conditional ablation of Msx1Msx2 in the mouse uterus using the PRcre mouse model. we isolated uterine stromal cells from day4 pregnant mice (n=5 for each genotype), purified total RNA from these cells, pooled these samples and then hybridized to high density affymetrix microarrays.本课题组的预实验显示，同源盒转录因子（homeobox transcription factors）Msx1与Msx2在小鼠妊娠早期的子宫组织中表达。进一步研究发现，在小鼠子宫中条件性敲除Msx1与Msx2，会因子宫上皮接受态受损而导致胚胎着床失败。为鉴定Msx1与Msx2在子宫内的下游靶基因，我们对妊娠第4天（胚胎着床窗口期）的Msx1Msx2双敲除小鼠及对应对照组的子宫基质细胞进行了基因表达谱分析。基因芯片（microarray）检测结果表明，在Msx1Msx2敲除小鼠的子宫基质中，成纤维细胞生长因子家族（fibroblast growth factor family）多个成员及Wnt家族（Wnts）的mRNA表达水平显著升高。本研究采用PR-Cre小鼠模型对小鼠子宫内的Msx1与Msx2实施条件性敲除：我们从妊娠第4天的小鼠体内分离子宫基质细胞（每个基因型组样本量为n=5），提取上述细胞的总RNA并混合样本，随后将混合样本与高密度Affymetrix基因芯片进行杂交。