Xiang Decai, Jia Baoyu, Guo Jianxiong, Shao Qingyong, Hong Qionghua, Wei Hongjiang, Quan Guobo, Wu Guoquan
Yunnan Provincial Genebank of Livestock and Poultry Genetic Resources, Yunnan Provincial Engineering Laboratory of Animal Genetic Resource Conservation and Germplasm Enhancement, Yunnan Animal Science and Veterinary Institute, Kunming, China.
Key Laboratory of Animal Gene Editing and Animal Cloning in Yunnan Province, College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China.
Front Genet. 2021 Dec 17;12:753327. doi: 10.3389/fgene.2021.753327. eCollection 2021.
Cryopreservation of porcine cloned zygotes has important implications for biotechnology and biomedicine research; however, lower embryo developmental potential remains an urgent problem to be resolved. For exploring the sublethal cryodamages during embryo development, this study was designed to acquire the mRNA and long non-coding RNA (lncRNA) profiles of 2-cells, 4-cells and blastocysts derived from vitrified porcine cloned zygotes using transcriptome sequencing. We identified 167 differentially expressed (DE) mRNAs and 516 DE lncRNAs in 2-cell stage, 469 DE mRNAs and 565 lncRNAs in 4-cell stage, and 389 DE mRNAs and 816 DE lncRNAs in blastocyst stage. Functional enrichment analysis revealed that the DE mRNAs during embryo development were involved in many regulatory mechanisms related to cell cycle, cell proliferation, apoptosis, metabolism and others. Moreover, the target genes of DE lncRNAs in the three embryonic stages were also enriched in many key GO terms or pathways such as "defense response", "linoleic acid metabolic process", "embryonic axis specification", "negative regulation of protein neddylation", etc., In conclusion, the present study provided comprehensive transcriptomic data about mRNAs and lncRNAs for the vitrified porcine cloned zygotes during different developmental stages, which contributed to further understand the potential cryodamage mechanisms responsible for impaired embryo development.
猪克隆受精卵的冷冻保存对生物技术和生物医学研究具有重要意义;然而,胚胎发育潜力较低仍是一个亟待解决的问题。为了探索胚胎发育过程中的亚致死性冷冻损伤,本研究旨在通过转录组测序获得源自玻璃化猪克隆受精卵的2细胞、4细胞和囊胚的mRNA和长链非编码RNA(lncRNA)图谱。我们在2细胞阶段鉴定出167个差异表达(DE)mRNA和516个DE lncRNA,在4细胞阶段鉴定出469个DE mRNA和565个lncRNA,在囊胚阶段鉴定出389个DE mRNA和816个DE lncRNA。功能富集分析表明,胚胎发育过程中的DE mRNA参与了许多与细胞周期、细胞增殖、凋亡、代谢等相关的调控机制。此外,三个胚胎阶段中DE lncRNA的靶基因也富集在许多关键的基因本体(GO)术语或通路中,如“防御反应”、“亚油酸代谢过程”、“胚胎轴特化”、“蛋白质NEDD化的负调控”等。总之,本研究提供了玻璃化猪克隆受精卵在不同发育阶段的mRNA和lncRNA的全面转录组数据,有助于进一步了解导致胚胎发育受损的潜在冷冻损伤机制。