Epigenetic reprogramming, gene expression and in vitro development of porcine SCNT embryos are significantly improved by a histone deacetylase inhibitor--m-carboxycinnamic acid bishydroxamide (CBHA).

Yuran Song,Liu Wang,Tang Hai,Ying Wang,Qi Zhou,Runfa Guo,Wei Li

doi:10.1007/s13238-014-0034-3

Abstract

Insufficient epigenetic reprogramming of donor nuclei is believed to be one of the most important causes of low development efficiency of mammalian somatic cell nuclear transfer (SCNT). Previous studies have shown that both the in vitro and in vivo development of mouse SCNT embryos could be increased significantly by treatment with various histone deacetylase inhibitors (HDACi), including Trichostatin A, Scriptaid, and m-carboxycinnamic acid bishydroxamide (CBHA), in which only the effect of CBHA has not yet been tested in other species. In this paper we examine the effect of CBHA treatment on the development of porcine SCNT embryos. We have discovered the optimum dosage and time for CBHA treatment: incubating SCNT embryos with 2 μmol/L CBHA for 24 h after activation could increase the blastocyst rate from 12.7% to 26.5%. Immunofluorescence results showed that the level of acetylation at histone 3 lysine 9 (AcH3K9), acetylation at histone 3 lysine 18 (AcH3K18), and acetylation at histone 4 lysine 16 (AcH4K16) was raised after CBHA treatment. Meanwhile, CBHA treatment improved the expression of development relating genes such as pou5f1, cdx2, and the imprinted genes like igf2. Despite these promising in vitro results and histone reprogramming, the full term development was not significantly increased after treatment. In conclusion, CBHA improves the in vitro development of pig SCNT embryos, increases the global histone acetylation and corrects the expression of some developmentally important genes at early stages. As in mouse SCNT, we have shown that nuclear epigenetic reprogramming in pig early SCNT embryos can be modified by CBHA treatment.Electronic supplementary materialThe online version of this article (doi:10.1007/s13238-014-0034-3) contains supplementary material, which is available to authorized users.

Highlights

As in mouse somatic cell nuclear transfer (SCNT), we have shown that nuclear epigenetic reprogramming in pig early SCNT embryos can be modified by carboxycinnamic acid bishydroxamide (CBHA) treatment
In vitro development of porcine SCNT embryos are improved by CBHA treatment To test whether CBHA could improve the early development of porcine SCNT embryos, we treated the SCNT embryos with different concentrations of CBHA and calculated the developmental rates from 2-cell to blastocyst stage (Table 1)
We found that proper CBHA treatment had no effect on the cleavage rates of porcine SCNT embryos, but could significantly increase the in vitro development rate to blastocyst stage

Summary

Introduction

Somatic cell nuclear transfer (SCNT) has been a successful technology to derive cloned animals in many species (Wilmut et al, 1997; Kato et al, 1998; Wakayama et al, 1998; Baguisi et al, 1999; Onishi et al, 2000; Chesne et al, 2002; Shin et al, 2002; Galli et al, 2003; Woods et al, 2003; Zhou et al, 2003; Lee et al, 2005; Li et al, 2006). Like in other animals, SCNT in pigs is inefficient, due to the low cloning efficiency, fetus abnormality, and placenta deficiency

Methods

Results

Conclusion