Département des Sciences Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, Québec, Québec G1V 0A6, Canada.
Reproduction. 2009 Jul;138(1):95-105. doi: 10.1530/REP-08-0533. Epub 2009 Apr 21.
Gene expression profiling is proving to be a powerful approach for the identification of molecular mechanisms underlying complex cellular functions such as the dynamic early embryonic development. The objective of this study was to perform a transcript abundance profiling analysis of bovine early embryonic development in vivo using a bovine developmental array. The molecular description of the first week of life at the mRNA level is particularly challenging when considering the important fluctuations in RNA content that occur between developmental stages. Accounting for the different intrinsic RNA content between developmental stages was achieved by restricting the reaction time during the global amplification steps and by using spiked controls and reference samples. Analysis based on intensity values revealed that most of the transcripts on the array were present at some point during in vivo bovine early embryonic development, while the varying number of genes detected in each developmental stage confirmed the dynamic profile of gene expression occurring during embryonic development. Pair-wise comparison of gene expression showed a marked difference between oocytes and blastocysts profiles, and principal component analysis revealed that the majority of the transcripts could be regrouped into three main clusters representing distinct RNA abundance profiles. Overall, these data provide a detailed temporal profile of the abundance of mRNAs revealing the richness of signaling processes in early mammalian development. Results presented here provide better knowledge of bovine in vivo embryonic development and contribute to the progression of our current knowledge regarding the first week of life in mammals.
基因表达谱分析被证明是一种强大的方法,可以用于识别复杂细胞功能(如动态早期胚胎发育)背后的分子机制。本研究的目的是使用牛发育数组对牛早期胚胎发育进行转录丰度谱分析。考虑到在发育阶段之间发生的 RNA 含量的重要波动,在 mRNA 水平上对生命的第一周进行分子描述特别具有挑战性。通过限制全局扩增步骤中的反应时间,并使用加标对照和参考样品,实现了对不同发育阶段内在 RNA 含量的考虑。基于强度值的分析表明,在体内牛早期胚胎发育的某个阶段,阵列上的大多数转录本都存在,而在每个发育阶段检测到的不同数量的基因证实了胚胎发育过程中发生的基因表达动态特征。基因表达的两两比较显示卵母细胞和囊胚之间的差异显著,主成分分析表明,大多数转录本可以分为三个主要簇,代表不同的 RNA 丰度谱。总体而言,这些数据提供了 mRNA 丰度的详细时间图谱,揭示了早期哺乳动物发育中信号转导过程的丰富性。本研究结果为牛体内胚胎发育提供了更好的认识,并有助于我们对哺乳动物生命第一周的现有知识的进展。