Transcriptome analysis of in vivo and in vitro matured bovine MII oocytes
Transcriptome analysis of in vivo and in vitro matured bovine MII oocytes
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DOI：
10.1016/j.theriogenology.2008.10.024
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发表时间：
2009-04-01
期刊：
影响因子：
2.8
通讯作者：
Gardner, D. K.
中科院分区：
文献类型：
--
作者：
Katz-Jaffe, M. G.;McCallie, B. R.;Gardner, D. K.
哺乳动物卵母细胞的体外成熟（IVM）与体内成熟的卵母细胞相比，不支持相同的胚胎发育或妊娠率。因此，环境对卵母细胞完成成熟并获得成功受精和正常胚胎发育所需的mRNA和蛋白质的能力有重要影响。本研究的目的是分析MII卵母细胞在体内和体外条件下的转录组。提取总RNA并将其与Affymetrix GeneChip (R)牛基因组阵列进行杂交，将芯片数据归一化后，分析发现与体内成熟对照相比，IVM后10个基因表达差异，包括Aqp3、Sept7、Abhd4和Siah2 （P < 0.05）。K-means聚类分析结合相关基因本体，确定了受IVM影响的几个生物过程，包括代谢、能量途径、细胞组织和生物发生以及细胞生长和维持。实时荧光定量PCR验证了微阵列数据，也显示了特异性推测印迹基因Igf2r、Peg3和Snrpn在IVM后表达水平的改变（P < 0.05）。不同的IVM转录模式反映了卵母细胞对周围环境的反应。监测关键卵母细胞成熟基因的转录水平可能随后有助于提高IVM的成功。(c) 2009爱思唯尔公司版权所有。
In vitro maturation (IVM) of mammalian oocytes does not support the same rates of embryo development or pregnancy when compared to oocytes that have matured in vivo. Therefore, environment has a significant influence on the oocyte's ability to complete maturation and acquire the mRNA and proteins required for successful fertilization and normal embryonic development. The aim of this study was to analyze the MII oocyte transcriptome between in vivo and in vitro conditions. Total RNA was extracted, processed and hybridized to the Affymetrix GeneChip (R) Bovine Genome Array, Following normalization of the microarray data, analysis revealed 10 differentially expressed genes after IVM compared to in vivo matured controls, including Aqp3, Sept7, Abhd4 and Siah2 (P < 0.05). K-means Cluster analysis coupled with associated gene ontology, identified several biological processes affected by IVM, including metabolism, energy pathways, cell organization and biogenesis, and cell growth and maintenance. Quantitative real-time PCR validated the microarray data and also revealed altered expression levels after IVM of specific putatively imprinted genes, Igf2r, Peg3 and Snrpn (P < 0.05). Distinct IVM transcription patterns reflected the oocyte's response to its surrounding environment. Monitoring transcription levels of key oocyte maturation genes may subsequently assist in improving IVM Success. (c) 2009 Elsevier Inc. All rights reserved.