The transcriptome of human oocytes
The transcriptome of human oocytes
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DOI：
10.1073/pnas.0603227103
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发表时间：
2006-09-19
影响因子：
11.1
通讯作者：
Cibelli, Jose Bernardo
中科院分区：
文献类型：
--
作者：
Kocabas, Arif Murat;Crosby, Javier;Cibelli, Jose Bernardo
从成熟的人类卵母细胞中识别基因和推断的途径可以帮助我们更好地了解卵子发生、卵泡发生、受精和胚胎发育。在取下卵巢后的几分钟内，使用人的M11卵母细胞，并将其转录组与由10个不同的正常人体组织(不包括卵巢)的总RNA组成的参考样本进行比较。使用一种独特的方法进行RNA扩增。用Affymetrix Human Genome U133 Plus 2.0基因芯片阵列进行杂交。与参考样本相比，卵母细胞中有5,331个转录本显著上调，7,074个转录本显著下调。在卵母细胞上调的探针中，有1,430个功能未知。通过将人类卵母细胞显著上调的基因与小鼠卵母细胞以及人和小鼠胚胎干细胞的基因相交，确定了66个转录本的核心组。基因芯片阵列的结果在一组选定的卵母细胞特异性基因中使用RT-PCR进行了验证。在上调的探针组中，表达最多的类别与RNA和蛋白质代谢有关，其次是DNA代谢和染色质修饰。这份报告提供了在体内成熟的人卵母细胞中表达的基因的全面表达基线。进一步了解这些基因的生物学作用可能会扩大我们对减数分裂细胞周期、受精、染色质重塑、谱系承诺、多能性、组织再生和形态发生的了解。
The identification of genes and deduced pathways from the mature human oocyte can help us better understand oogenesis, folliculogenesis, fertilization, and embryonic development. Human metaphase 11 oocytes were used within minutes after removal from the ovary, and its transcriptome was compared with a reference sample consisting of a mixture of total RNA from 10 different normal human tissues not including the ovary. RNA amplification was performed by using a unique protocol. Affymetrix Human Genome U133 Plus 2.0 GeneChip arrays were used for hybridizations. Compared with reference samples, there were 5,331 transcripts significantly up-regulated and 7,074 transcripts significantly down-regulated in the oocyte. Of the oocyte up-regulated probe sets, 1,430 have unknown function. A core group of 66 transcripts was identified by intersecting significantly up-regulated genes of the human oocyte with those from the mouse oocyte and from human and mouse embryonic stem cells. GeneChip array results were validated using RT-PCR in a selected set of oocyte-specific genes. Within the up-regulated probe sets, the top overrepresented categories were related to RNA and protein metabolism, followed by DNA metabolism and chromatin modification. This report provides a comprehensive expression baseline of genes expressed in in vivo matured human oocytes. Further understanding of the biological role of these genes may expand our knowledge on meiotic cell cycle, fertilization, chromatin remodeling, lineage commitment, pluripotency, tissue regeneration, and morphogenesis.