title: Effects of histone deacetylase inhibitor oxamflatin on in vitro porcine somatic cell nuclear transfer embryos.
abstract: Low cloning efficiency is considered to be caused by the incomplete or aberrant epigenetic reprogramming of differentiated donor cells in somatic cell nuclear transfer (SCNT) embryos. Oxamflatin, a novel class of histone deacetylase inhibitor (HDACi), has been found to improve the in vitro and full-term developmental potential of SCNT embryos. In the present study, we studied the effects of oxamflatin treatment on in vitro porcine SCNT embryos. Our results indicated that the rate of in vitro blastocyst formation of SCNT embryos treated with 1 μM oxamflatin for 15 h postactivation was significantly higher than all other treatments. Treatment of oxamflatin decreased the relative histone deacetylase (HDAC) activity in cloned embryos and resulted in hyperacetylation levels of histone H3 at lysine 9 (AcH3K9) and histone H4 at lysine 5 (AcH4K5) at pronuclear, two-cell, and four-cell stages partly through downregulating HDAC1. The suppression of HDAC6 through oxamflatin increased the nonhistone acetylation level of α-tubulin during the mitotic cell cycle of early SCNT embryos. In addition, we demonstrated that oxamflatin downregulated DNA methyltransferase 1 (DNMT1) expression and global DNA methylation level (5-methylcytosine) in two-cell-stage porcine SCNT embryos. The pluripotency-related gene POU5F1 was found to be upregulated in the oxamflatin-treated group with a decreased DNA methylation tendency in its promoter regions. Treatment of oxamflatin did not change the locus-specific DNA methylation levels of Sus scrofa heterochromatic satellite DNA sequences at the blastocyst stage. Meanwhile, our findings suggest that treatment with HDACi may contribute to maintaining the stable status of cytoskeleton-associated elements, such as acetylated α-tubulin, which may be the crucial determinants of donor nuclear reprogramming in early SCNT embryos. In summary, oxamflatin treatment improves the developmental potential of porcine SCNT embryos in vitro.
authors: 1 Key Laboratory of Agriculture Animal Genetics, Breeding and Reproduction, College of Animal Science, Huazhong Agricultural University , Wuhan, 430070, China .
authors: Liming
authors: L
authors: Hou
authors: Fanhua
authors: F
authors: Ma
authors: Jinzeng
authors: J
authors: Yang
authors: Hasan
authors: H
authors: Riaz
authors: Yongliang
authors: Y
authors: Wang
authors: Wangjun
authors: W
authors: Wu
authors: Xiaoliang
authors: X
authors: Xia
authors: Zhiyuan
authors: Z
authors: Ma
authors: Ying
authors: Y
authors: Zhou
authors: Lin
authors: L
authors: Zhang
authors: Wenqin
authors: W
authors: Ying
authors: Dequan
authors: D
authors: Xu
authors: Bo
authors: B
authors: Zuo
authors: Zhuqing
authors: Z
authors: Ren
authors: Yuanzhu
authors: Y
authors: Xiong
mesh_terms: D000107:Acetylation / Q000187:drug effects; D000818:Animals; D019976:Cloning, Organism; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000096:biosynthesis; D019175:DNA Methylation / Q000187:drug effects; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D056284:Histone Deacetylase 1 / Q000096:biosynthesis; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006657:Histones / Q000378:metabolism; D006877:Hydroxamic Acids / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D034421:Sus scrofa; D014404:Tubulin / Q000378:metabolism
chemical_list: D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; D050814:Octamer Transcription Factor-3; D014404:Tubulin; C100692:oxamflatin; D004248:DNA (Cytosine-5-)-Methyltransferases; D056284:Histone Deacetylase 1