Suberoylanilide hydroxamic acid, a novel histone deacetylase inhibitor, improves the development and acetylation level of miniature porcine handmade cloning embryos

Abstract

Histone deacetylase inhibitors (HDACis) can change the histone acetylation and significantly enhance the developmental competence of the pre-implantation SCNT embryo. To select a proper histone deacetylase inhibitor to improve the success rate and potentially developmental ability of handmade cloning (HMC) embryos of miniature porcine, we compared the effect of two histone deacetylase inhibitors (SAHA vs. VPA) on HMC embryo development, their histone acetylation level and the expression level of relevant genes. The blastocyst rate and number of blastocyst cells of HMC embryos treated with SAHA (SAHA-HMC) or VPA (VPA-HMC) were significantly higher than those of control (Control-HMC), respectively, but there were no significant difference between SAHA-HMC and VPA-HMC groups. In addition, the acetylation level (AcH4K8) of Control-HMC and VPA-HMC embryos at the blastocyst stage, respectively, was significantly lower than that of in vitro fertilized (IVF) and SAHA-HMC embryos. However, the acetylation H4K8 of the blastocysts had no significant difference between SAHA-HMC and the IVF groups. The SAHA-HMC blastocysts indicated comparative expression levels of Oct4 and HDAC1 (histone deacetyltransferase gene) with those of IVF blastocysts. In contrast, the expression levels of Oct4 were lower and those of HDAC1 were higher in the VPA-HMC and Control-HMC blastocysts, respectively, compared to those of the IVF blastocysts. Our results demonstrated that the HMC embryos treated by SAHA could promote the pre-implantation development and increase the levels of histone H4K8 acetylation and the expression of the OCT4 gene, yet decrease the expression of the HDAC1 gene to the comparable level of the IVF embryos. Our results proved that SAHA may be a better histone deacetylase inhibitor for porcine HMC compared to VPA, and furthermore, it may indicate that SAHA can effectively correct the abnormal histone acetylation during the HMC embryo development and subsequently improve the full-term developmental potential of the HMC embryos after embryo transplantation.

Epigenetic Features of Animal Biotechnologies

“…Analyses performed on imprinted genes also showed improved reprogramming after HDACi treatments: in the porcine SCNT protocols, scriptaid treatment of the donor cells rescued disrupted methylation at H19 imprinted region after SCNT and oxamflatin treatment improved the DNA methylation profile at the imprinted XIST locus that lead to higher in vitro developmental rates and offspring rates (Xu et al 2013;Hou et al 2014). Similar results were obtained on developmental key genes Oct4, Nanog and Sox2 (Huan et al 2015;Jin et al 2017;Sun et al 2017).…”

Functional role of increased acetylated tubulin in porcine oocyte microtubule structure, meiotic maturation and embryogenesis

“…Oocyte maturation process and formula follow our previous experiments (36). Cumulus oocyte complexes (COCs) were cultured in the maturation medium containing 15 IU/mL eCG and 10 IU/mL hCG for 20-22h, and transferred to free hormone maturation medium for the remaining 20-22h.…”

“…Fertilization process and formula follow our previous experiments (36). The matured cumulus was gently blown and digested.…”

“…Reverse transcription and QRT-PCR procedures were performed according to our previous study (36). Comparison of relative expression levels of microtubule assembly genes.…”

HDAC6 Is Involved in the Histone Deacetylation of In Vitro Maturation Oocytes and the Reprogramming of Nuclear Transplantation in Pig

“…The N-terminal acetylation of histones H3 and H4 is an important part of epigenetic modification and nuclear transplantation [1]. The H4K8 acetylation of pseudopronucleus is close to that of male and female pronucleus, which upregulates the expression of pluripotent genes and improves the efficiency of embryo development [1,2]. Histone H4K5 and H3K9/K14 acetylation are involved in the protamine replacement and the regulation of chromosome structure [3,4].…”