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）

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 increase...

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Published in:蛋白质与细胞：英文版 2014 (5), p.382-393
Main Author: Yuran Song Tang Hai Ying Wang Runfa Guo Wei Li Liu Wang Qi Zhou
Format: Article
Language:English
Subjects:
体外发育
体细胞核移植胚胎
去乙酰化
基因表达
组蛋白乙酰化
育猪
遗传修饰
酶抑制剂
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Summary: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 treat- ment with various histone deacetylase inhibitors （HDACi）, including Trichostatin A, Scriptaid, and m-carboxycin- namic 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 dis- covered the optimum dosage and time for CBHA treat- ment： incubating SCNT embryos with 2 pmol/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 pou5fl, cdx2, and the imprinted genes like igf2. Despite these promising in vitro results and histone reprogramming, the full term development was notsignificantly 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.
ISSN:1674-800X
1674-8018