Effects of Scriptaid on Cell Cycle and Histone Acetylation of Ovine Nuclear Donor Cumulus Cells and their Ability to Support the Development of Somatic Cell Nuclear Transfer Embryos
Compelling evidence suggests that histone deacetylase inhibitor (HDACi) influences the development of somatic cell nuclear transfer (SCNT) embryos. The current study was conducted to determine the effect of pretreatment of donor cumulus cells with Scriptaid (a novel HDACi) on cell cycle, histone acetylation and cloning embryos development in ovine. First, we optimized the efficiency of Scriptaid in a dose (0, 0.1, 0.2, 0.4 and 0.8 μmol/L) and time-dependent (0, 12, 24, 36, and 48 h) manner on the developmental capacity of these embryos. Then, we quantitatively assessed the alterations of acetylation levels in histone H3 lysine 9 (acH3K9) and histone H4 lysine 12 (acH4K12) of cumulus cells and SCNT embryos by immunofluorescence staining. Furthermore, we detected the proportion of G0/G1 phase cells in cumulus cells. We found a significantly improved blastocyst development rates of cloning embryos derived from donor cumulus cells pretreated with a mild dose (0.2 μmol/L) of Scriptaid for 24 hours (21/86 [24.39%] vs. 11/85 [12.91%]; P<0.05). Meanwhile, the levels of acH3K9 and acH4K12 were also improved significantly in cumulus cells and SCNT embryos (P<0.05). Moreover, more cumulus cells pretreated with Scriptaid were in G0/G1 phase compared with control group (84.22% vs. 75.96%, P<0.05). In conclusion, donor cumulus cells treated with Scriptaid is beneficial to early development of SCNT embryos, ascending acH3K9/ acH4K12 and G0/G1 phase cells proportion of cumulus cell. Scriptaid can be used to improve the efficiency of somatic cell nuclear transfer in ovine.
Hui Cao, Wenlong Su, Junjie Li, Bingqiang Wen, Shujun Tian, Shuchun Sun, Lu Li, Hanyang Wang and Jiexin Li