Effects of Trichostatin A on In Vitro Development and DNA Methylation Level of the Satellite I
Region of Swamp Buffalo (Bubalus bubalis) Cloned Embryos |
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Authors: | Kanokwan SRIRATTANA Mariena KETUDAT-CAIRNS Takashi NAGAI Masahiro KANEDA Rangsun PARNPAI |
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Institution: | 1)Embryo Technology and Stem Cell Research Center and School of Biotechnology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand;2)NARO National Institute of Livestock and Grassland Science, Ibaraki 305-0901, Japan;3)Division of Animal Life Science, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan |
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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. |
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Keywords: | Cloning DNA methylation Embryo development Swamp buffalo Trichostatin A |
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