Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.
Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.
Theriogenology. 2022 Jan 1;177:140-150. doi: 10.1016/j.theriogenology.2021.10.017. Epub 2021 Oct 20.
It has been reported that N6-methyladenosine (m6A) methyltransferase-like 3 (METTL3) plays an important role in zygote genome activation during embryonic development, but the effects of METTL3 under oxidative stress in the early development of goat embryos remain largely unknown. In this study, zygotes were monitored at 72 and 168 h after fertilization, and they developed to the 8-cell stage and blastocyst stage under hypoxic conditions and normoxic conditions. Single-cell transcriptome sequencing was performed at the 8-cell stage and the blastocyst stage in the goat embryos, the differentially expressed METTL3 was screened from the sequencing results. We found that microinjection of small interfering RNA (siRNA) against METTL3 caused developmental arrest, both 8-cell rates (37.45 ± 2.21% vs. 47.09 ± 1.38%; P < 0.01) and blastocyst rates of Si-METTL3 (12.17% ± 2.84 vs. 20.83 ± 3.61%; P < 0.01) in Si-METTL3 group were significantly decreased compared with that of control under hypoxic conditions, significant changes were found in the m6A-related genes and the expression levels of critical transcription factors, such as, NANOG, GATA3, CDX2 and SOX17, were decreased. This study revealed the key role of METTL3 in the regulation of embryonic development under oxidative stress, and laid the foundation for further study of the crucial mechanism of oxidative stress during the early embryonic development of goats.
据报道，N6-甲基腺苷（m6A）甲基转移酶样 3（METTL3）在胚胎发育过程中的合子基因组激活中发挥重要作用，但 METTL3 在山羊胚胎早期发育过程中的氧化应激下的作用在很大程度上仍不清楚。在这项研究中，在受精后 72 和 168 小时监测受精卵，在缺氧和常氧条件下它们发育到 8 细胞期和囊胚期。对山羊胚胎的 8 细胞期和囊胚期进行单细胞转录组测序，从测序结果中筛选出差异表达的 METTL3。我们发现，针对 METTL3 的小干扰 RNA（siRNA）的微注射导致发育停滞，在缺氧条件下，8 细胞率（37.45±2.21%比 47.09±1.38%；P<0.01）和 Si-METTL3 的囊胚率（12.17%±2.84%比 20.83%±3.61%；P<0.01）明显低于对照组，m6A 相关基因的表达水平和关键转录因子（如 NANOG、GATA3、CDX2 和 SOX17）的表达水平均显著降低。这项研究揭示了 METTL3 在氧化应激下调节胚胎发育的关键作用，为进一步研究山羊早期胚胎发育过程中氧化应激的关键机制奠定了基础。
