title: Combination of S-adenosylhomocysteine and scriptaid, a non-toxic epigenetic modifying reagent, modulates the reprogramming of bovine somatic-cell nuclear transfer embryos.
abstract: The goal of this study was to improve the development of bovine somatic-cell nuclear transfer (SCNT) embryos by optimizing the combination of DNA methyltransferases inhibitor S-adenosylhomocysteine (SAH) and histone deacetylase inhibitor Scriptaid (SPD). A. 4 × 4-factor design of different drug combinations (0, 0.75, 1.0, and 1.5 mM SAH and 0, 5, 250, and 500 nM SPD) was used to identify an optimal combination of 0.75 mM SAH and 250 nM SPD that improved the developmental competence of bovine SCNT embryos. Further experiments using this combination revealed that methylation levels of CpG islands near exon 1 of the pluripotent gene SOX2; the epigenetic-related gene HDAC3 and DNMT3a; imprinted genes XIST and PEG3; as well as apoptosis-related genes BCL2 and BAX were returned to levels similar to those of in vitro fertilized (IVF) embryo after treatment, which also normalized transcript levels for these genes. This combination also returned global DNA methylation to a normal level, correcting H4K12ac levels while enhancing H3K9ac levels. Thus, the combined application of 0.75 mM SAH and 250 nM SPD can significantly improve the reprogramming of bovine SCNT embryos by stabilizing how embryos utilize their genomes.
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi Province, People's Republic of China; Key Laboratory of Biological Technology, Ministry of Agriculture, Yangling, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Wang
authors.forename: Yuankun
authors.initials: Y
authors.lastname: Sang
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Song
authors.initials: S
authors.lastname: Hua
mesh_terms: D000818:Animals; D001483:Base Sequence; D001947:Breeding / Q000379:methods*; D002417:Cattle / Q000196:embryology*; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation / Q000187:drug effects; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors; D017931:DNA Primers / Q000235:genetics; D004338:Drug Combinations; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000196:embryology; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005455:Fluorescent Antibody Technique; D006652:Histological Techniques; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006655:Histone Deacetylases / Q000235:genetics; D006898:Hydroxylamines / Q000494:pharmacology*; D008969:Molecular Sequence Data; D053652:Nuclear Transfer Techniques; D011804:Quinolines / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012435:S-Adenosylhomocysteine / Q000494:pharmacology*; D055748:SOXB1 Transcription Factors / Q000235:genetics; D017422:Sequence Analysis, DNA; D000099186:Histone Deacetylase 3
chemical_list: D015254:DNA Modification Methylases; D017931:DNA Primers; D004338:Drug Combinations; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases; D006898:Hydroxylamines; D011804:Quinolines; D012435:S-Adenosylhomocysteine; D055748:SOXB1 Transcription Factors; D000099186:Histone Deacetylase 3; C410733:scriptaid