Abstract：
Vitrification of porcine immature oocytes at the germinal vesicle (GV) stage reduces subsequent embryo yield and changes at the molecular level may occur during embryonic development. Therefore, the present study used porcine parthenogenetic embryos as a model to investigate the effect of GV oocyte vitrification on the transcriptional profiles of the resultant embryos at the 4-cell and blastocyst stages using the Smart-seq2 RNA-seq technique. We identified 743 (420 up-regulated and 323 down-regulated) and 994 (554 up-regulated and 440 down-regulated) differentially expressed genes (DEGs) from 4-cell embryos and blastocysts derived from vitrified GV oocytes, respectively. Functional enrichment analysis of DEGs in 4-cell embryos showed that vitrification of GV oocytes influenced regulatory mechanisms related to transcription regulation, apoptotic process, metabolism and key pathways such as the MAPK signaling pathway. Moreover, DEGs in blastocysts produced from vitrified GV oocytes were enriched in critical biological functions including cell adhesion, cell migration, AMPK signaling pathway, GnRH signaling pathway and so on. In addition, the transcriptomic analysis and quantitative real-time PCR results were consistent. In summary, the present study revealed that the vitrification of porcine GV oocytes could alter gene expression patterns during subsequent embryonic developmental stages, potentially affecting their developmental competence.
摘要：
在生发囊泡（GV）阶段，猪未成熟卵母细胞的玻璃化会降低随后的胚胎产量，并且在胚胎发育过程中可能会发生分子水平的变化。因此,本研究使用猪孤雌生殖胚胎作为模型，使用Smart-seq2RNA-seq技术研究GV卵母细胞玻璃化对4细胞和胚泡阶段所得胚胎转录谱的影响。我们从玻璃化GV卵母细胞的4细胞胚胎和胚泡中鉴定出743（420上调和323下调）和994（554上调和440下调）差异表达基因（DEG），分别。4细胞胚胎中DEGs的功能富集分析表明，GV卵母细胞的玻璃化影响与转录调节相关的调节机制，凋亡过程，代谢和关键通路，如MAPK信号通路。此外，玻璃化GV卵母细胞产生的胚泡中的DEGs富含关键的生物学功能，包括细胞粘附，细胞迁移，AMPK信号通路,GnRH信号通路等。此外,转录组学分析和实时定量PCR结果一致。总之,本研究表明，猪GV卵母细胞的玻璃化可以在随后的胚胎发育阶段改变基因表达模式，可能会影响他们的发展能力。
