title: Epigenetic Modification Agents Improve Gene-Specific Methylation Reprogramming in Porcine Cloned Embryos.
abstract: Incomplete DNA methylation reprogramming in cloned embryos leads to poor cloning efficiency. Epigenetic modification agents can improve genomic methylation reprogramming and the development of cloned embryos, however, the effect of epigenetic modification agents on gene-specific methylation reprogramming remains poorly studied. Here, we investigated DNA methylation reprogramming of pluripotency (Oct4) and tissue specific (Thy1) genes during early embryo development in pigs. In this study, we found that compared with in vitro fertilized counterparts, cloned embryos displayed the disrupted patterns of Oct4 demethylation and Thy1 remethylation. When 5-aza-2'-deoxycytidine (5-aza-dC) or trichostatin A (TSA) enhanced the development of cloned embryos, the transcripts of DNA methyltransferases (Dnmt1 and Dnmt3a), histone acetyltransferase 1 (Hat1) and histone deacetylase 1 (Hdac1) and the methylation and expression patterns of Oct4 and Thy1 became similar to those detected in in vitro fertilized counterparts. Further studies showed that Dnmt1 knockdown in cloned embryos enhanced the methylation reprogramming of Oct4 and Thy1 and promoted the activation of Oct4 and the silence of Thy1. In conclusion, our results demonstrated that cloned embryos displayed incomplete gene-specific methylation reprogramming and disrupted expression patterns of pluripotency and tissue specific genes, and epigenetic modification agents improved gene-specific methylation reprogramming and expression pattern by regulating epigenetic modification related genes. This work would have important implications in improving cloning efficiency.
authors.affiliation: College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, Shandong Province, China.
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Huan
authors.affiliation: Shouguang City Hospital of Chinese Medicine, Weifang, Shandong Province, China.
authors.forename: Zhanfeng
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Shouguang City Hospital of Chinese Medicine, Weifang, Shandong Province, China.
authors.forename: Jiguang
authors.initials: J
authors.lastname: Zhang
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang Province, China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhu
authors.affiliation: College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang Province, China.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Liu
authors.affiliation: College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, Shandong Province, China.
authors.forename: Xuexiong
authors.initials: X
authors.lastname: Song
mesh_terms: D000818:Animals; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000187:drug effects*; D019175:DNA Methylation / Q000187:drug effects*; D000077209:Decitabine; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006877:Hydroxamic Acids / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine / Q000196:embryology* / Q000235:genetics*
chemical_list: D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D012333:RNA, Messenger; C012589:trichostatin A; D000077209:Decitabine; D001374:Azacitidine