Expression of eukaryotic elongation initiation factor 1A differentially marks zygotic genome activation in biparental and parthenogenetic porcine embryos and correlates with in vitro developmental potential
Expression of eukaryotic elongation initiation factor 1A differentially marks zygotic genome activation in biparental and parthenogenetic porcine embryos and correlates with in vitro developmental potential
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DOI：
10.1071/rd08072
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发表时间：
2008-01-01
影响因子：
1.9
通讯作者：
Cabot, Ryan A.
中科院分区：
文献类型：
--
作者：
Magnani, Luca;Johnson, Christine M.;Cabot, Ryan A.
合子基因组激活（ZGA）是卵裂发育过程中的一个重要事件。哺乳动物胚胎的体外操作（包括胚胎培养）可能导致ZGA前后的发育停滞。真核细胞延伸起始因子1A（eIF 1A）已被用作某些哺乳动物物种中ZGA的标记。我们假设eIF 1A的表达可用于评估猪的ZGA;我们还假设eIF 1A的表达谱可用于评估体外发育潜力。本研究的目的是确定eIF 1A在猪卵裂发育过程中的表达模式，并评估其在不同质量胚胎中的表达水平。我们使用真实的时间逆转录-聚合酶链反应测定定量eIF 1A转录本在不同的时间点在卵裂发育过程中产生的猪胚胎孤雌激活（PA）和体外受精（IVF）。我们发现eIF 1A在IVF胚胎的两细胞期和PA胚胎的四细胞期被激活。我们发现，在孤雌胚胎中观察到的转录水平的增加是依赖于从头转录。我们发现，与同一时间点的四细胞胚胎相比，激活后48小时的孤雌胚胎中的eIF 1A转录水平发生了变化，表现为两个或八个细胞胚胎。我们的工作支持了eIF 1A是猪ZGA的标志物的假设，其表达谱可用于评估胚胎质量。
Zygotic genome activation (ZGA) is a major event during cleavage development. In vitro manipulation of mammalian embryos (including embryo culture) can result in developmental arrest around the time of ZGA. Eukaryotic elongation initiation factor 1A (eIF1A) has been used as a marker for ZGA in some mammalian species. We hypothesised expression of eIF1A can be used to assess ZGA in the pig; we also hypothesised that the expression profile of eIF1A can be used to assess developmental potential in vitro. The aims of the present study were to determine the expression pattern of eIF1A during porcine cleavage development and to assess its expression levels in embryos of different quality. We used a real- time reverse transcription-polymerase chain reaction assay to quantify eIF1A transcripts at different time points during cleavage development in porcine embryos produced by parthenogenetic activation (PA) and in vitro fertilisation (IVF). We found that eIF1A is activated at the two-cell stage in IVF embryos and at the four-cell stage in PA embryos. We showed that the increase in transcript levels observed in parthenogenetic embryos is dependent on de novo transcription. We found altered levels of eIF1A transcripts in parthenogenetic embryos that presented as either two-or eight-cell embryos 48 h after activation compared with four-cell embryos at the same time point. Our work supports the hypothesis that eIF1A is a marker of porcine ZGA and its expression profile can be used to assess embryo quality.