Highly sensitive sequencing reveals dynamic modifications and activities of small RNAs in mouse oocytes and early embryos.
Highly sensitive sequencing reveals dynamic modifications and activities of small RNAs in mouse oocytes and early embryos.
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DOI：
10.1126/sciadv.1501482
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发表时间：
2016-06
期刊：
影响因子：
13.6
通讯作者：
Wu L
中科院分区：
文献类型：
--
作者：
Yang Q;Lin J;Liu M;Li R;Tian B;Zhang X;Xu B;Liu M;Zhang X;Li Y;Shi H;Wu L
敏感测序揭示了早期小鼠胚胎中microRNAs的表达、修饰和活性的动态调节。小RNA在早期胚胎发育中起着重要作用。然而，由于可用于测序分析的RNA稀缺，人们对它们的表达动态和修饰知之甚少。使用一种改进的深度测序方法，只需要10 ng总RNA或50个卵母细胞，我们就可以分析小鼠卵母细胞和早期胚胎中的小RNA。我们发现，microRNA(MiRNA)在受精后不久就开始表达，与卵母细胞相比，精子在受精过程中携带到受精卵中的成熟miRNAs相对较少。有趣的是，在单细胞到双细胞胚胎中，受精卵miRNAs的3‘-单腺苷酸化和寡腺苷化显著增加，这可能保护miRNAs免受在此期间发生的大规模降解。此外，生物信息学分析表明，miRNA的功能从卵母细胞到两细胞阶段受到抑制，在两细胞阶段后似乎重新激活，以调节在胚胎发育中重要的基因。因此，我们的研究为进一步检测早期胚胎中的小RNA提供了一种高度敏感的图谱方法和有价值的数据集。
Expression, modification, and activity of microRNAs are dynamically regulated in early mouse embryos unveiled by sensitive sequencing. Small RNAs play important roles in early embryonic development. However, their expression dynamics and modifications are poorly understood because of the scarcity of RNA that is obtainable for sequencing analysis. Using an improved deep sequencing method that requires as little as 10 ng of total RNA or 50 oocytes, we profile small RNAs in mouse oocytes and early embryos. We find that microRNA (miRNA) expression starts soon after fertilization, and the mature miRNAs carried into the zygote by sperm during fertilization are relatively rare compared to the oocyte miRNAs. Intriguingly, the zygotic miRNAs display a marked increase in 3′ mono- and oligoadenylation in one- to two-cell embryos, which may protect the miRNAs from the massive degradation taking place during that time. Moreover, bioinformatics analyses show that the function of miRNA is suppressed from the oocyte to the two-cell stage and appears to be reactivated after the two-cell stage to regulate genes important in embryonic development. Our study thus provides a highly sensitive profiling method and valuable data sets for further examination of small RNAs in early embryos.