ELF4 is critical to zygotic gene activation and epigenetic reprogramming during early embryonic development in pigs

Abstract

Zygotic gene activation (ZGA) and epigenetic reprogramming are critical in early embryonic development in mammals, and transcription factors are involved in regulating these events. However, the effects of ELF4 on porcine embryonic development remain unclear. In this study, the expression of ELF4 was detected in early porcine embryos and different tissues. By knocking down ELF4, the changes of H3K9me3 modification, DNA methylation and ZGA-related genes were analyzed. Our results showed that ELF4 was expressed at all stages of early porcine embryos fertilized in vitro (IVF), with the highest expression level at the 8-cell stage. The embryonic developmental competency and blastocyst quality decreased after ELF4 knockdown (20.70% control vs. 17.49% si-scramble vs. 2.40% si-ELF4; p < 0.001). Knockdown of ELF4 induced DNA damage at the 4-cell stage. Interfering with ELF4 resulted in abnormal increases in H3K9me3 and DNA methylation levels at the 4-cell stage and inhibited the expression of genes related to ZGA. These results suggest that ELF4 affects ZGA and embryonic development competency in porcine embryos by maintaining genome integrity and regulating dynamic changes of H3K9me3 and DNA methylation, and correctly activating ZGA-related genes to promote epigenetic reprogramming. These results provide a theoretical basis for further studies on the regulatory mechanisms of ELF4 in porcine embryos.

Cumulus Cell and Oocyte Gene Expression in Prepubertal Gilts and Sows Identifies Cumulus Cells as a Prime Informative Parameter of Oocyte Quality

“…A panel of reference genes, previously validated and/or used as reliable reference genes in similar studies, namely Actin beta (ACTB) [27,[48][49][50][51], glyceraldehyde-3-phosphate dehydrogenase (GAPDH) [27,46,49,52], hydroxymethylbilane synthase (HMBS aka PBGD) [53][54][55], hypoxanthine phosphoribosyltransferase 1 (HPRT1) [56] and tyrosine 3-Monooxygenase/ Tryptophan 5-Monooxygenase Activation Protein Gamma (YWHAG) [49] were compared using NormFinder [57], which identified ACTB as the most stable reference gene for both CCs and oocytes. The transcript abundance from each gene was hence normalized against the abundance of the reference transcript ACTB.…”