UniGene cDNA array-based monitoring of transcriptome changes during placental development mouse
UniGene cDNA array-based monitoring of transcriptome changes during placental development mouse
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DOI：
10.1073/pnas.231396598
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发表时间：
2001-11-06
影响因子：
11.1
通讯作者：
Himmelbauer, H
中科院分区：
文献类型：
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作者：
Hemberger, M;Cross, JC;Himmelbauer, H
胎盘是胚胎生长和发育所必需的高度特化的器官。在这里，我们应用了早期（妊娠第7.5天）和晚期胚胎（第17.5天）的胚外组织之间的cDNA减法，以产生阶段特异性cDNA池，用于筛选含有25，000个克隆的高密度小鼠UniGene cDNA阵列。共鉴定了638个克隆，488个具有e7.5特异性探针，150个具有e17.5特异性探针。重要的是，363/638（56.9%）的杂交克隆是未知的胎盘发育过程中表达之前。通过北方印迹和原位杂交证实了总共44/44个阳性克隆的差异调节。因此，这种组合的cDNA减法和阵列杂交是非常成功的胎盘发育过程中表达和调控的基因的鉴定。这些包括生长因子和受体，转录和翻译机制的组成部分，细胞周期调节因子，分子伴侣和细胞骨架元件。广泛的原位杂交分析显示胚胎外结构与高密度的差异表达的基因，最引人注目的是胎盘外锥和海绵滋养层。这种大规模的胎盘发生过程中调控的基因鉴定是非常有用的，以进一步阐明胚外发育的分子基础。
The placenta is a highly specialized organ essential for embryonic growth and development. Here, we have applied cDNA subtraction between extraembryonic tissues of early- (day 7.5 of gestation) and late-stage embryos (day 17.5) to generate stage-specific cDNA pools that were used for screening of high-density mouse UniGene cDNA arrays containing 25,000 clones. A total of 638 clones were identified, 488 with the e7.5-specific probe and 150 with the e17.5-specific probe. Importantly, 363/638 (56.9%) of the hybridizing clones were not known to be expressed during placental development before. Differential regulation was confirmed by Northern blot and in situ hybridization for a total of 44, /44 of positive clones. Thus, this combination of cDNA subtraction and array hybridization was highly successful for identification of genes expressed and regulated during placental development. These included growth factors and receptors, components of the transcriptional and translational machinery, cell cycle regulators, molecular chaperones, and cytoskeletal elements. The extensive in situ hybridization analysis revealed extraembryonic structures with a high density of differentially expressed genes, most strikingly the ectoplacental cone and the spongiotrophoblast. This large-scale identification of genes regulated during placentogenesis is extremely useful to further elucidate the molecular basis of extraembryonic development.