title: Alteration of the DNA methylation status of donor cells impairs the developmental competence of porcine cloned embryos.
abstract: Nuclear reprogramming induced by somatic cell nuclear transfer is an inefficient process, and donor cell DNA methylation status is thought to be a major factor affecting cloning efficiency. Here, the role of donor cell DNA methylation status regulated by 5-aza-2'-deoxycytidine (5-aza-dC) or 5-methyl-2'-deoxycytidine-5'-triphosphate (5-methyl-dCTP) in the early development of porcine cloned embryos was investigated. Our results showed that 5-aza-dC or 5-methyl-dCTP significantly reduced or increased the global methylation levels and altered the methylation and expression levels of key genes in donor cells. However, the development of cloned embryos derived from these cells was reduced. Furthermore, disrupted pseudo-pronucleus formation and transcripts of early embryo development-related genes were observed in cloned embryos derived from these cells. In conclusion, our results demonstrated that alteration of the DNA methylation status of donor cells by 5-aza-dC or 5-methyl-dCTP disrupted nuclear reprogramming and impaired the developmental competence of porcine cloned embryos.
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Yan Jun
authors.initials: YJ
authors.lastname: Huan
authors.forename: Zhan Feng
authors.initials: ZF
authors.lastname: Wu
authors.forename: Ji Guang
authors.initials: JG
authors.lastname: Zhang
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhu
authors.forename: Bing Teng
authors.initials: BT
authors.lastname: Xie
authors.forename: Jian Yu
authors.initials: JY
authors.lastname: Wang
authors.forename: Jing Yu
authors.initials: JY
authors.lastname: Li
authors.forename: Bing Hua
authors.initials: BH
authors.lastname: Xue
authors.forename: Qing Ran
authors.initials: QR
authors.lastname: Kong
authors.forename: Zhong Hua
authors.initials: ZH
authors.lastname: Liu
mesh_terms: D000818:Animals; D001374:Azacitidine / Q000031:analogs & derivatives / Q000737:chemistry; D001755:Blastocyst / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D019976:Cloning, Organism / Q000379:methods*; D019175:DNA Methylation; D000077209:Decitabine; D003845:Deoxycytosine Nucleotides / Q000737:chemistry; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects; D005307:Fertilization in Vitro; D005347:Fibroblasts / Q000378:metabolism; D016678:Genome; D009865:Oocytes / Q000166:cytology; D013552:Swine
chemical_list: D003845:Deoxycytosine Nucleotides; C034817:5-methyldeoxycytidine triphosphate; D000077209:Decitabine; D001374:Azacitidine