Dynamic Change Patterns of H3K64ac in Early Development of Porcine (Sus scrofa domesticus) Somatic Cell Cloned Embryos
ZHANG Dan-Dan, XU Teng-Teng, GAO Di, NING Wei, QI Xin, RU Zhen-Yuan, ZHANG Xiang-Dong, LI Yun-Sheng, ZHANG Yun-Hai, CAO Zu-Bing*
Anhui Provincial Key Laboratory of Livestock and Poultry Genetic Resources Protection and Biological breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
Abstract：Histone acetylation is the first histone modification to be identified, which regulates gene expression by changing chromatin structure and accessibility, and is widely involved in the regulation of various biological events during early embryonic development of mammals. H3K64ac is a previously unidentified histone modification on the lateral surface of the histone octamer. It is known to be involved in regulating nucleosome stability and promoting gene expression in vivo. However, the role of H3K64ac in the early development of porcine (Sus scrofa domesticus) somatic nuclear transfer embryos is still unknown. In this study, H3K64ac in early embryos derived from parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) were examined by immunofluorescence staining. PA 4-cell embryos were treated with aphidicolin, α-amanitin and cycloheximide, respectively. The results showed that H3K64ac modification was always present in both PA and SCNT embryos, and H3K64ac levels were the maximum at the pronuclear stage, decreased at the 2-cell stage, slightly increased at the 4-cell stage, and remained low status from the 8-cell stage to the blastocyst stage. Comparison analyses of H3K64ac levels between PA and SCNT embryos revealed that H3K64ac levels in SCNT embryos at each stage were significantly higher than those in PA embryos (P<0.05). Inhibition of DNA replication, RNA and protein synthesis did not affect H3K64ac levels in porcine embryos. The results suggested that H3K64ac was abnormally reprogramming during early development of porcine cloned embryos, and the reduction of H3K64ac levels in early embryonic development was an active reactive process independent of DNA replication, RNA and protein synthesis. This study provides a theoretical basis for further study on the molecular mechanism of H3K64ac involved in regulating the development of porcine somatic nuclear transfer embryos.
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