Miyoshi Kazuchika, Mori Hironori, Mizobe Yamato, Akasaka Eri, Ozawa Akio, Yoshida Mitsutoshi, Sato Masahiro
Laboratory of Animal Reproduction, Faculty of Agriculture, Kagoshima University, Korimoto, Kagoshima, Japan.
Cell Reprogram. 2010 Feb;12(1):67-74. doi: 10.1089/cell.2009.0032.
The present study was carried out to examine the effects of valproic acid (VPA), a histone deacetylase inhibitor, on in vitro development of miniature pig somatic cell nuclear transfer (SCNT) embryos and on expression of a mouse Oct-3/4 promoter-driven enhanced green fluorescent protein (EGFP) gene (EGFP expression only detected in Oct-3/4-expressing cells) introduced into donor cells for SCNT during their development. The addition of 4 mM VPA to embryo culture medium for 48 h after activation significantly (p < 0.01) increased the blastocyst formation rate of SCNT embryos compared with the control, whereas VPA did not affect their cleavage rate. The rate of SCNT embryos expressing EGFP at 5 days of culture was not affected by the presence or absence of VPA treatment. At 7 days of culture, however, the addition of 4 mM VPA to embryo culture medium for 48 h after activation significantly (p < 0.05) increased the rate of SCNT embryos expressing EGFP compared with the control. The results indicate that VPA enhances the ability of miniature pig SCNT embryos to develop into blastocysts and maintains the ability of them to express Oct-3/4 gene.
本研究旨在检测组蛋白去乙酰化酶抑制剂丙戊酸(VPA)对小型猪体细胞核移植(SCNT)胚胎体外发育的影响,以及对在SCNT供体细胞发育过程中导入的小鼠Oct-3/4启动子驱动的增强型绿色荧光蛋白(EGFP)基因(仅在表达Oct-3/4的细胞中检测到EGFP表达)表达的影响。激活后在胚胎培养基中添加4 mM VPA 48小时,与对照组相比,显著(p < 0.01)提高了SCNT胚胎的囊胚形成率,而VPA不影响其分裂率。培养5天时,有无VPA处理对SCNT胚胎表达EGFP的比率没有影响。然而,在培养7天时,激活后在胚胎培养基中添加4 mM VPA 48小时,与对照组相比,显著(p < 0.05)提高了SCNT胚胎表达EGFP的比率。结果表明,VPA增强了小型猪SCNT胚胎发育成囊胚的能力,并维持了它们表达Oct-3/4基因的能力。