Effect of vitrification on the microRNA transcriptome in mouse blastocysts.
Effect of vitrification on the microRNA transcriptome in mouse blastocysts.
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玻璃化冷冻对小鼠囊胚中 MicroRNA 转录组的影响
DOI：
10.1371/journal.pone.0123451
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Zhu H
中科院分区：
文献类型：
--
作者：
Zhao X;Hao H;Du W;Zhu H
玻璃化冷冻是哺乳动物囊胚冷冻保存中常用的方法，以克服胚胎移植的时间和空间限制。先前的研究表明，玻璃化冷冻胚泡的着床能力受到了损害，而microRNAs(MiRNAs)调控着床的关键基因。然而，关于玻璃化冷冻对囊胚miRNA转录组的影响的信息很少。本研究采用基于miRNA Taqman分析的方法对新鲜和玻璃化冷冻的小鼠囊胚的miRNA转录本进行了分析，并用实时定量聚合酶链式反应(qRT-PCR)对结果进行了验证。然后，使用基因本体论(GO)和京都基因和基因组百科全书(KEGG)数据库对差异表达的miRNAs进行评估。总体而言，在玻璃化冷冻和新鲜的小鼠囊胚中检测到了760个已知的小鼠miRNAs。其中，5种miRNAs的表达水平差异显著：在玻璃化冷冻囊胚中，4种miRNAs(MMU-miR-199a-5p、MMU-miR-329-3p、MMU-miR-136-5p和MMU-miR-16-1-3p)表达上调，而1种miRNAs(MMU-miR-212-3p)表达下调。基于miRNA Taqman法和qRT-PCR法检测的所有miRNAs的表达水平均一致。上调的4个miRNAs可调控877个候选靶基因，下调的miRNA可调控231个基因。生物学分析进一步表明，差异表达的miRNAs主要调控胚胎着床。综上所述，我们的研究结果表明，玻璃化冷冻显著改变了小鼠囊胚的miRNA转录组，这可能会降低玻璃化冷冻囊胚的着床潜力。
Vitrification is commonly used in the cryopreservation of mammalian blastocysts to overcome the temporal and spatial limitations of embryo transfer. Previous studies have shown that the implantation ability of vitrified blastocysts is impaired and that microRNAs (miRNAs) regulate the critical genes for embryo implantation. However, little information is available about the effect of vitrification on the miRNA transcriptome in blastocysts. In the present study, the miRNA transcriptomes in fresh and vitrified mouse blastocysts were analyzed by miRNA Taqman assay based method, and the results were validated using quantitative real-time PCR (qRT-PCR). Then, the differentially expressed miRNAs were assessed using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Overall, 760 known mouse miRNAs were detected in the vitrified and fresh mouse blastocysts. Of these, the expression levels of five miRNAs differed significantly: in the vitrified blastocysts, four miRNAs (mmu-miR-199a-5p, mmu-miR-329-3p, mmu-miR-136-5p and mmu-miR-16-1-3p) were upregulated, and one (mmu-miR-212-3p) was downregulated. The expression levels of all miRNAs measured by the miRNA Taqman assay based method and qRT-PCR were consistent. The four upregulated miRNAs were predicted to regulate 877 candidate target genes, and the downregulated miRNA was predicted to regulate 231 genes. The biological analysis further showed that the differentially expressed miRNAs mainly regulated the implantation of embryos. In conclusion, the results of our study showed that vitrification significantly altered the miRNA transcriptome in mouse blastocysts, which may decrease the implantation potential of vitrified blastocysts.
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影响因子：
3.1
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通讯作者：
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