犏牛体外受精胚胎的发育转录组分析

doi:10.11843/j.issn.0366-6964.2018.01.011

摘要/Abstract

摘要：

本研究旨在探讨犏牛早期胚胎发育调控机制，为提高犏牛胚胎体外生产效率提供理论基础。以娟珊牛精子体外受精（IVF）牦牛卵母细胞获得的2-、4-、8-细胞、桑椹胚和囊胚5个阶段的杂种胚胎分别提取总RNA，采用Smart-Seq2扩增、构建测序文库，应用RNA-Seq技术进行高通量测序分析。对HiSeq^TM2500测序所得的Raw Reads进行数据过滤后，得到5个发育阶段的犏牛早期胚胎的Clean Reads为47 791 850~63 332 216条，有80.00%~91.13%比对上参考基因的序列。4-细胞期表达的基因数最多（15 400），而桑椹胚表达的基因数最少（9 604）。以|log₂ ratio|≥ 1，Q-value<0.05为阈值，获得2-~4-细胞、4-~8-细胞、8-细胞~桑椹胚及桑椹胚~囊胚4个发育阶段的差异表达基因（DEGs）分别为3 690、6 332、8 965和10 298个。GO分析表明，4个发育阶段的DEGs归类注释都涉及生物过程（BP）、细胞组分（CC）和分子功能（MF）3大类67个二级条目。KEGG分析表明，牦牛早期胚胎发育过程中DEGs富集的主要通路有剪接体（Spliceosome）、神经活性配体-受体互作（Neuroactive ligand-receptor interaction）、细胞因子及其受体相互作用（Cytokine-cytokine receptor interactions）、泛素介导的蛋白水解（Ubiquitin mediated proteolysis）、RNA转运（RNA transport）等通路。本研究利用RNA-Seq技术对犏牛体外受精胚胎在发育不同阶段转录组进行了分析，获得了众多差异基因和有关通路的富集，为完善犏牛胚胎体外生产技术提供了理论基础。

Abstract:

The objective of this study was to investigate the regulatory mechanism of the early development of hybrid embryos of the yak (Bos grunniens) in order to provide theoretical basis for improving efficiency of in vitro production (IVP). Total RNA were extracted from embryos that derived from yak oocytes in vitro fertilized with Jersey cattle at 5 stages of 2-cell, 4-cell, 8-cell, morula and blastocyst and then amplified via the Smart-Seq2 method to construct RNA libraries and sequenced using RNA-seq. A total of 47 791 850-63 332 216 Clean Reads for each in vitro stage after filtering the Raw Reads of HiSeq^TM2500 sequence were obtained, of which, 80.00% to 91.13% Reads were covered in the reference genome. Number of expressed genes were greatest at 4-cell stage (15 400) and lowest at blastocyst stage (9 604). When|log₂ ratio| ≥ 1 and Q-value<0.05 were set as thresholds for identifying differentially expressed genes (DEGs), a total of 3 690, 6 332, 8 965 and 10 298 DEGs were identified for 2-cell-4 cell, 4-cell-8-cell, 8-cell-morula and morula-blastocyst, respectively. The GO distributions of the DEGs for 4 stages were classified into 3 categories:biological processes (BP), cellular components (CC), molecular functions (MF) with a total of 67 subcategories.KEGG enrichment analysis of DEGs showed that the major enrichment pathways were Spliceosome, Neuroactive ligand-receptor interaction, Cytokine-cytokine receptor interactions, Ubiquitin mediated proteolysis, RNA transport etc. In conclusion, this was the report to investigate the mechanism that the early development of hybrid in vitro-derived yak embryos using high-throughput sequencing. A number of DEGs and pathways were discovered, which provided valuable information to improve in vitro production technique for yak crossbred embryos.

中图分类号:

S823.8⁺5.2

字向东, 刘霜, 夏威, 熊显荣, 黄林, 张正帆, 李志雄, 李键, 钟金城, 王利, 朱江江. 犏牛体外受精胚胎的发育转录组分析[J]. 畜牧兽医学报, 2018, 49(1): 92-101.

ZI Xiang-dong, LIU Shuang, XIA Wei, XIONG Xian-rong, HUANG Lin, ZHANG Zheng-fan, LI Zhi-xiong, LI Jian, ZHONG Jin-cheng, WANG Li, ZHU Jiang-jiang. Transcriptome Analysis during Development of Hybrid in vitro-derived Yak Embryos[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(1): 92-101.

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https://www.xmsyxb.com/CN/Y2018/V49/I1/92

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