Abstract
Abstract
Use of histone deacetylase inhibitors (HDACis) is believed to improve the developmental competence and quality of cloned embryos produced. We examined the effects of treatment of buffalo fibroblasts with valproic acid (VPA), a HDACi on these cells and on embryos produced from them by hand-made cloning. VPA treatment (1.5, 3.0, or 4.5 mM) altered (p < 0.05) the growth characteristics and relative expression level of HDAC1, DNMT1, DNMT3a, P53, and CASPASE3, and the global level of H3K9/14ac, H4K5ac, and H3K18ac but not H3K27me3 in the cells. After the use of VPA-treated donor cells for producing embryos, the cleavage and blastocyst rate, and total cell number were not significantly affected; however, the apoptotic index was lower (p < 0.05) for 3.0 or 4.5 mM VPA group than for 1.5 mM VPA group or the controls. In the cloned blastocysts, the expression level of HDAC1 was higher (p < 0.05) and CASPASE3 was lower (p < 0.05), whereas that of DNMT1, DNMT3a, and P53 and the global level of H3K9/14ac were not significantly affected after VPA treatment of donor cells. In conclusion, these results suggest that VPA treatment of donor cells adversely affects their growth characteristics, increases histone acetylation, and alters gene expression but does not improve production rate of cloned embryos.
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