Kocabas Arif Murat, Crosby Javier, Ross Pablo J, Otu Hasan H, Beyhan Zeki, Can Handan, Tam Wai-Leong, Rosa Guilherme J M, Halgren Robert G, Lim Bing, Fernandez Emilio, Cibelli Jose Bernardo
Cellular Reprogramming Laboratory, Department of Animal Science, Michigan State University, East Lansing, MI 48824, USA.
Proc Natl Acad Sci U S A. 2006 Sep 19;103(38):14027-32. doi: 10.1073/pnas.0603227103. Epub 2006 Sep 12.
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 II 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.
从成熟人类卵母细胞中鉴定基因和推导相关途径，有助于我们更好地理解卵子发生、卵泡发生、受精及胚胎发育过程。人类减数分裂中期II卵母细胞在从卵巢取出后的数分钟内被使用，其转录组与一个参考样本进行了比较，该参考样本由来自10种不同的不包括卵巢的正常人类组织的总RNA混合物组成。通过使用独特的方案进行RNA扩增。使用Affymetrix人类基因组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5331个转录本显著上调，7074个转录本显著下调。在卵母细胞上调的探针组中，有1430个功能未知。通过将人类卵母细胞、小鼠卵母细胞以及人类和小鼠胚胎干细胞中显著上调的基因进行交叉比对，鉴定出了一个由66个转录本组成的核心组。使用RT-PCR在一组选定的卵母细胞特异性基因中对基因芯片阵列结果进行了验证。在上调的探针组中，最主要的类别与RNA和蛋白质代谢相关，其次是DNA代谢和染色质修饰。本报告提供了在体内成熟的人类卵母细胞中表达的基因的全面表达基线。对这些基因生物学作用的进一步了解可能会扩展我们对减数分裂细胞周期、受精、染色质重塑、细胞谱系定向分化、多能性、组织再生和形态发生的认识。
