Department of Molecular Biology, Cellular Biology and Biochemistry, Brown University, Providence, RI 02912, USA.
Dev Biol. 2012 Feb 1;362(1):104-13. doi: 10.1016/j.ydbio.2011.11.015. Epub 2011 Dec 3.
microRNAs (miRNAs) are small noncoding RNAs that mediate post-transcriptional gene regulation and have emerged as essential regulators of many developmental events. The transcriptional network during early embryogenesis of the purple sea urchin, Strongylocentrotus purpuratus, is well described and can serve as an excellent model to test functional contributions of miRNAs in embryogenesis. We examined the loss of function phenotypes of major components of the miRNA biogenesis pathway. Inhibition of de novo synthesis of Drosha and Dicer in the embryo led to consistent developmental defects, a failure to gastrulate, and embryonic lethality, including changes in the steady state levels of transcription factors and signaling molecules involved in germ layer specification. We annotated and profiled small RNA expression from the ovary and several early embryonic stages by deep sequencing followed by computational analysis. miRNAs as well as a large population of putative piRNAs (piwi-interacting RNAs) had dynamic accumulation profiles through early development. Defects in morphogenesis caused by loss of Drosha could be rescued with four miRNAs. Taken together our results indicate that post-transcriptional gene regulation directed by miRNAs is functionally important for early embryogenesis and is an integral part of the early embryonic gene regulatory network in S. purpuratus.
微小 RNA(miRNA)是一种小型非编码 RNA,可介导转录后基因调控,并且已成为许多发育事件的重要调控因子。紫色海胆(Strongylocentrotus purpuratus)早期胚胎发生的转录网络已得到很好的描述,可作为测试 miRNA 在胚胎发生中功能贡献的极佳模型。我们检查了 miRNA 生物发生途径的主要成分丧失功能的表型。在胚胎中抑制 Drosha 和 Dicer 的从头合成会导致一致的发育缺陷,原肠胚形成失败以及胚胎致死,包括涉及胚层特化的转录因子和信号分子的稳态水平的变化。我们通过深度测序和计算分析,对卵巢和几个早期胚胎阶段的小 RNA 表达进行注释和分析。miRNA 以及大量假定的 piRNA(piwi 相互作用 RNA)在早期发育过程中具有动态积累谱。由 Drosha 缺失引起的形态发生缺陷可以通过四种 miRNA 进行挽救。总之,我们的结果表明,miRNA 指导的转录后基因调控对于早期胚胎发生是功能上重要的,并且是 S. purpuratus 早期胚胎基因调控网络的组成部分。