Porcine zygotic genome activation (ZGA) occurs along with global epigenetic remodeling at the 4-cell stage. These processes are regulated by histone acetylation, which requires acetyl-coenzyme A (CoA). Pyruvate dehydrogenase complex (PDC) is a crucial enzyme in glucose metabolism that converts pyruvate into acetyl-CoA. In mammalian cells, acetyl-CoA is produced by pyruvate dehydrogenase alpha 1 (PDHA1) translocated into the nucleus in special conditions. To determine whether zygotic PDHA1 plays a critical role in promoting histone acetylation during ZGA, a CRISPR/Cas9 genome editing system using multiple guide RNAs was employed to generate a PDHA1-targeted parthenogenetic embryo model. Results of immunofluorescent staining showed that the nuclear accumulation of PDHA1 during ZGA was significantly inhibited by PDHA1 targeting. Meanwhile, the 4-cell arrest rate significantly increased at 72 h after activation, indicating impeded embryonic development. In addition, nuclear histone acetylation significantly decreased when PDHA1 was targeted, and quantitative PCR showed that expression of several zygotic genes was significantly decreased in the PDHA1-targeting group compared to the control group. Overexpression of PDHA1 recovered the nuclear PDHA1, H3K9Ac and H3K27Ac and EIF1A expression levels. Moreover, the 5-to-8-cell-stage embryo development rate was only partially rescued. In conclusion, expression of zygotic origin PDHA1 contributes to porcine ZGA by maintaining histone acetylation in porcine embryos.

中文翻译：

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丙酮酸脱氢酶α1的核积累促进组蛋白乙酰化，并且对于猪胚胎中的合子基因组激活是必不可少的。

猪合子基因组激活（ZGA）与4细胞阶段的全局表观遗传重塑一起发生。这些过程受组蛋白乙酰化的调节，这需要乙酰辅酶A（CoA）。丙酮酸脱氢酶复合物（PDC）是葡萄糖代谢中的关键酶，可将丙酮酸转化为乙酰CoA。在哺乳动物细胞中，乙酰辅酶A是由丙酮酸脱氢酶α1（PDHA1）在特殊条件下转移到细胞核中产生的。为了确定合子PDHA1是否在促进ZGA期间促进组蛋白乙酰化中起关键作用，采用了使用多个引导RNA的CRISPR / Cas9基因组编辑系统来生成靶向PDHA1的孤雌生殖胚胎模型。免疫荧光染色的结果表明，PDHA1靶向显着抑制了ZGA期间PDHA1的核积累。与此同时，活化后72小时，4-细胞阻滞率显着增加，表明胚胎发育受阻。此外，当靶向PDHA1时，核组蛋白乙酰化显着降低，定量PCR显示，与对照组相比，靶向PDHA1的组中一些合子基因的表达显着降低。PDHA1的过表达恢复了核PDHA1，H3K9Ac和H3K27Ac和EIF1A的表达水平。此外，仅部分挽救了5至8细胞期胚胎的发育速度。总之，合子来源PDHA1的表达通过维持猪胚胎中的组蛋白乙酰化而有助于猪ZGA。靶向PDHA1时，核组蛋白乙酰化显着降低，定量PCR显示，与对照组相比，靶向PDHA1的组中一些合子基因的表达显着降低。PDHA1的过表达恢复了核PDHA1，H3K9Ac和H3K27Ac和EIF1A的表达水平。此外，仅部分挽救了5至8细胞期胚胎的发育速度。总之，合子来源PDHA1的表达通过维持猪胚胎中的组蛋白乙酰化而有助于猪ZGA。靶向PDHA1时，核组蛋白乙酰化显着降低，定量PCR显示，与对照组相比，靶向PDHA1的组中一些合子基因的表达显着降低。PDHA1的过表达恢复了核PDHA1，H3K9Ac和H3K27Ac和EIF1A的表达水平。此外，仅部分挽救了5至8细胞期胚胎的发育速度。总之，合子来源PDHA1的表达通过维持猪胚胎中的组蛋白乙酰化而有助于猪ZGA。5至8个细胞阶段的胚胎发育率仅部分得以挽救。总之，合子来源PDHA1的表达通过维持猪胚胎中的组蛋白乙酰化而有助于猪ZGA。5至8个细胞阶段的胚胎发育率仅部分得以挽救。总之，合子来源PDHA1的表达通过维持猪胚胎中的组蛋白乙酰化而有助于猪ZGA。