Silva P G C, Moura M T, Silva R L O, Nascimento P S, Silva J B, Ferreira-Silva J C, Cantanhêde L F, Chaves M S, Benko-Iseppon A M, Oliveira M A L
2Laboratório de Biotécnicas Reprodutivas,Departamento de Medicina Veterinária,Universidade Federal Rural de Pernambuco,Brasil.
1Laboratório de Biotécnicas Reprodutivas,Departamento de Medicina Veterinária,Universidade Federal Rural de Pernambuco,Brasil.
Zygote. 2018 Aug;26(4):270-278. doi: 10.1017/S0967199418000175. Epub 2018 Jul 23.
SummaryPluripotency-associated transcription factors (PATFs) modulate gene expression during early mammalian embryogenesis. Despite a strong understanding of PATFs during mouse embryogenesis, limited progress has been made in ruminants. This work aimed to describe the temporal expression of eight PATFs during both sheep and cattle preimplantation development. Transcript availability of PATFs was evaluated by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) in eggs, cleavage-stage embryos, morulae, and blastocysts. Transcripts of five genes were detected in all developmental stages of both species (KLF5, OCT4, RONIN, ZFP281, and ZFX). Furthermore, CMYC was detected in all cattle samples but was found from cleavage-stage onwards in sheep. In contrast, NR0B1 was detected in all sheep samples but was not detected in cattle morulae. GLIS1 displayed the most significant variation in temporal expression between species, as this PATF was only detected in cattle eggs and sheep cleavage-stage embryos and blastocysts. In silico analysis suggested that cattle and sheep PATFs share similar size, isometric point and molecular weight. A phenetic analysis showed two patterns of PATF clustering between cattle and sheep, among several mammalian species. In conclusion, the temporal expression of pluripotency-associated transcription factors differs between sheep and cattle, suggesting species-specific regulation during preimplantation development.
摘要
多能性相关转录因子（PATFs）在哺乳动物早期胚胎发育过程中调节基因表达。尽管对小鼠胚胎发育过程中的PATFs有深入了解，但在反刍动物方面进展有限。本研究旨在描述绵羊和牛着床前发育过程中8种PATFs的时间表达情况。通过逆转录定量聚合酶链反应（RT-qPCR）评估卵、卵裂期胚胎、桑葚胚和囊胚中PATFs的转录本可用性。在两个物种的所有发育阶段均检测到5个基因的转录本（KLF5、OCT4、RONIN、ZFP281和ZFX）。此外，在所有牛样本中均检测到CMYC，但在绵羊中从卵裂期开始才检测到。相反，在所有绵羊样本中均检测到NR0B1，但在牛桑葚胚中未检测到。GLIS1在物种间的时间表达变化最为显著，因为该PATF仅在牛卵以及绵羊卵裂期胚胎和囊胚中检测到。计算机分析表明，牛和绵羊的PATFs具有相似的大小、等电点和分子量。聚类分析显示，在几种哺乳动物物种中，牛和绵羊的PATFs存在两种聚类模式。总之，绵羊和牛的多能性相关转录因子的时间表达存在差异，这表明在着床前发育过程中存在物种特异性调控。
