牦牛新鲜囊胚与玻璃化冻融囊胚转录组的比较分析

doi:10.11843/j.issn.0366-6964.2018.04.007

摘要/Abstract

摘要：

旨在探讨牦牛在囊胚玻璃化冷冻前后转录组差异表达，明确玻璃化冷冻对牦牛囊胚基因表达谱的影响。采用体外受精生产的牦牛新鲜囊胚和玻璃化冻融囊胚为研究对象，分别提取总RNA，使用Smart-seq2方法进行扩增、构建文库、转录组测序（RNA-seq），以|log₂（冻胚/鲜胚）|≥1和Q<0.05为阈值筛选差异表达基因（DEG），并对DEGs进行GO功能富集和KEGG分析。结果表明，冷冻前后牦牛囊胚中分别检测到9 827和13 567个转录本。在两个文库共筛选出11 174个DEGs，其中有7 037个在冻融囊胚上调，4 137个下调。有10 538个DEGs显著富集（P<0.05）到479条GO terms上，共涉及318条通路，其中真核生物的核糖体合成、剪接体和神经活性配体-受体互作等8条通路显著富集（P<0.05）。本研究利用RNA-seq技术分析了牦牛囊胚在玻璃化冷冻前后转录组变化，为探讨牦牛囊胚冷冻损伤机制及进一步完善冷冻方法提供理论依据。

Abstract:

The study aimed to investigate the transcriptome differential expression between pre-and post-vitrified yak blastocysts in order to understand the effect of vitrification on the gene expression profile of yak blastocysts. Total RNAs were extracted respectively from fresh blastocysts (FRB) and vitrified-thawed blastocysts (VTB) that were in vitro-produced from yaks, and then the Smart-seq2 method was used to amplify and construct RNA libraries, and finally transcriptome was sequenced using RNA-seq. The|log₂ (VTB/FRB)| ≥ 1 and Q-value<0.05 were set as thresholds for identifying differentially expressed gene (DEG). The DEGs were then searched against the GO and KEGG database for enrichment analysis. The results showed that 9 827 and 13 567 transcripts were detected in FRB and VTB, respectively. A total of 11 174 DEGs were identified between the 2 libraries, in which 7 037 were up-regulated, and 4 137 were down-regulated in VTB. 10 538 DEGs were significantly enriched in 479 GO terms (P<0.05), and there were 318 pathways involved, of which 8 pathways including ribosome biogenesis in eukaryotes, spliceosome and neuroactive ligand-receptor interaction were significantly enriched (P<0.05). In conclusion, this report analyzed the transcriptome change pre-and post-vitrification of yak blastocysts using RNA-seq, which provides a theoretical basis to explore the mechanism of vitrification damage and improve vitrification technology of yak blastocysts.

中图分类号:

S823.8⁺5.2

蒲思颖, 郑杰, 杨远潇, 王琴, 杨绕芬, 字向东. 牦牛新鲜囊胚与玻璃化冻融囊胚转录组的比较分析[J]. 畜牧兽医学报, 2018, 49(4): 709-717.

PU Si-ying, ZHENG Jie, YANG Yuan-xiao, WANG Qin, YANG Rao-fen, ZI Xiang-dong. Comparative Transcriptome Analysis between Fresh and Vitrified-thawed Blastocysts of the Yak (Bos grunniens)[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(4): 709-717.

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https://www.xmsyxb.com/CN/Y2018/V49/I4/709

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