title: Effects of trichostatin A on In vitro development and DNA methylation level of the satellite I region of swamp buffalo (Bubalus bubalis) cloned embryos.
abstract: Trichostatin A (TSA), a histone deacetylase inhibitor, has been widely used to improve the cloning efficiency in several species. This brings our attention to investigation of the effects of TSA on developmental potential of swamp buffalo cloned embryos. Swamp buffalo cloned embryos were produced by electrical pulse fusion of male swamp buffalo fibroblasts with swamp buffalo enucleated oocytes. After fusion, reconstructed oocytes were treated with 0, 25 or 50 nM TSA for 10 h. The results showed that there was no significant difference in the rates of fusion (82-85%), cleavage (79-84%) and development to the 8-cell stage (59-65%) among treatment groups. The highest developmental rates to the morula and blastocyst stages of embryos were found in the 25 nM TSA-treated group (42.7 and 30.1%, respectively). We also analyzed the DNA methylation level in the satellite I region of donor cells and in in vitro fertilized (IVF) and cloned embryos using the bisulfite DNA sequencing method. The results indicated that the DNA methylation levels in cloned embryos were significantly higher than those of IVF embryos but approximately similar to those of donor cells. Moreover, there was no significant difference in the methylation level among TSA-treated and untreated cloned embryos. Thus, TSA treatments at 25 nM for 10 h could enhance the in vitro developmental potential of swamp buffalo cloned embryos, but no beneficial effect on the DNA methylation level was observed.
authors.affiliation: Embryo Technology and Stem Cell Research Center and School of Biotechnology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand.
authors.forename: Kanokwan
authors.initials: K
authors.lastname: Srirattana
authors.forename: Mariena
authors.initials: M
authors.lastname: Ketudat-Cairns
authors.forename: Takashi
authors.initials: T
authors.lastname: Nagai
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kaneda
authors.forename: Rangsun
authors.initials: R
authors.lastname: Parnpai
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000737:chemistry / Q000502:physiology; D002020:Buffaloes / Q000196:embryology* / Q000235:genetics; D002459:Cell Fusion / Q000662:veterinary; D019976:Cloning, Organism; D004247:DNA / Q000032:analysis; D019175:DNA Methylation / Q000187:drug effects*; D004276:DNA, Satellite / Q000378:metabolism; D046149:Embryo Culture Techniques / Q000662:veterinary; D004622:Embryo, Mammalian / Q000737:chemistry; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D005347:Fibroblasts / Q000737:chemistry; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008297:Male; D009028:Morula / Q000737:chemistry / Q000502:physiology; D053652:Nuclear Transfer Techniques / Q000662:veterinary
chemical_list: D004276:DNA, Satellite; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; C012589:trichostatin A; D004247:DNA