Hwang In-Sul, Shim Joohyun, Oh Keon Bong, Lee Haesun, Park Mi-Ryung
Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Wanju 55365, Jeonbuk, Korea.
Columbia Center for Translational Immunology, Department of Medicine, Columbia University Irving Medical Center, New York, NY 10032, USA.
Animals (Basel). 2022 Jun 28;12(13):1662. doi: 10.3390/ani12131662.
cd26 is ubiquitously distributed in the body, particularly in the endothelial and epithelial cells, with the highest expression in the kidney, liver, and small intestine. In humans, cd26 serves as a marker for the embryo implantation phase. However, little is known about the role of cd26 in porcine pre-implantation embryo development. Here, we aimed to examine siRNA-induced cd26 downregulation in the cytoplasm of MII oocytes, to determine whether cd26 is involved in the regulation of porcine pre-implantation embryonic development. The cd26 siRNA was micro-injected into the cytoplasm of MII oocytes, which were then parthenogenetically activated electrically in a medium containing 0.3M Mannitol. Inhibition of the cd26 expression did not affect cleavage but stopped development in the blastocyst stage. Additionally, the cd26 siRNA-treated blastocysts had significantly more apoptotic cells than the untreated blastocysts. Among the 579 transcripts evaluated with transcriptome resequencing, 38 genes were differentially expressed between the treatment and control blastocysts (p < 0.05). Twenty-four genes were upregulated in cd26 siRNA-injected blastocysts, whereas 14 were downregulated. These genes are involved in apoptosis, accumulation of reactive oxygen species, and aberrant expression of ribosomal protein genes. Our results indicate that cd26 is required for proper porcine parthenogenetic activation during embryonic development.
CD26在体内广泛分布,尤其在内皮细胞和上皮细胞中,在肾脏、肝脏和小肠中表达最高。在人类中,CD26作为胚胎着床期的一个标志物。然而,关于CD26在猪着床前胚胎发育中的作用知之甚少。在此,我们旨在检测小干扰RNA(siRNA)诱导的MII期卵母细胞胞质中CD26表达下调情况,以确定CD26是否参与猪着床前胚胎发育的调控。将CD26 siRNA显微注射到MII期卵母细胞的胞质中,然后在含有0.3M甘露醇的培养基中对其进行电孤雌激活。抑制CD26表达不影响卵裂,但阻止胚胎发育至囊胚阶段。此外,经CD26 siRNA处理的囊胚比未处理的囊胚有明显更多的凋亡细胞。在用转录组重测序评估的579个转录本中,处理组和对照组囊胚之间有38个基因差异表达(p<0.05)。在注射CD26 siRNA的囊胚中,24个基因上调,14个基因下调。这些基因参与细胞凋亡、活性氧积累以及核糖体蛋白基因的异常表达。我们的结果表明,CD26是猪胚胎发育过程中正常孤雌激活所必需的。