Corcoran D, Fair T, Park S, Rizos D, Patel O V, Smith G W, Coussens P M, Ireland J J, Boland M P, Evans A C O, Lonergan P
School of Agriculture, Food Science and Veterinary Medicine and Centre for Integrative Biology, Conway Institute for Biomolecular and Biomedical Research, College of Life Sciences, University College Dublin, Belfield, Dublin 4, Ireland.
Reproduction. 2006 Apr;131(4):651-60. doi: 10.1530/rep.1.01015.
In vivo-derived bovine embryos are of higher quality than those derived in vitro. Many of the differences in quality can be related to culture environment-induced changes in mRNA abundance. The aim of this study was to identify a range of mRNA transcripts that are differentially expressed between bovine blastocysts derived from in vitro versus in vivo culture. Microarray (BOTL5) comparison between in vivo- and in vitro-cultured bovine blastocysts identified 384 genes and expressed sequence tags (ESTs) that were differentially expressed; 85% of these were down-regulated in in vitro cultured blastocysts, showing a much reduced overall level of mRNA expression in in vitro- compared with in vivo-cultured blastocysts. Relative expression of 16 out of 23 (70%) differentially expressed genes (according to P value) were verified in new pools of in vivo- and in vitro-cultured blastocysts, using quantitative real-time PCR. Most (10 out of 16) are involved in transcription and translation events, suggesting that the reason why in vitro-derived embryos are of inferior quality compared with in vivo-derived embryos is due to a deficiency of the machinery associated with transcription and translation.
体内来源的牛胚胎比体外来源的胚胎质量更高。许多质量差异可能与培养环境引起的mRNA丰度变化有关。本研究的目的是鉴定一系列在体外培养与体内培养的牛囊胚之间差异表达的mRNA转录本。对体内培养和体外培养的牛囊胚进行微阵列（BOTL5）比较，鉴定出384个差异表达的基因和表达序列标签（EST）；其中85%在体外培养的囊胚中表达下调，表明与体内培养的囊胚相比，体外培养的囊胚中mRNA表达的总体水平大幅降低。使用定量实时PCR在新的体内培养和体外培养囊胚样本中验证了23个差异表达基因（根据P值）中的16个（70%）的相对表达。大多数（16个中的10个）参与转录和翻译过程，这表明体外来源的胚胎质量不如体内来源的胚胎的原因是与转录和翻译相关的机制存在缺陷。
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