Zhao Jianguo, Hao Yanhong, Ross Jason W, Spate Lee D, Walters Eric M, Samuel Melissa S, Rieke August, Murphy Clifton N, Prather Randall S
National Swine Resource and Research Center, University of Missouri, Columbia, Missouri 65211, USA.
Cell Reprogram. 2010 Feb;12(1):75-83. doi: 10.1089/cell.2009.0038.
Faulty epigenetic reprogramming of somatic nuclei is likely to be a major cause of low success observed in all mammals produced through somatic cell nuclear transfer (SCNT). It has been demonstrated that the developmental competence of SCNT embryos in several species were significantly enhanced via treatment of histone deacetylase inhibitors (HDACi) such as trichostatin A (TSA) to increase histone acetylation. Here we report that 50 nM TSA for 10 h after activation increased the developmental competence of porcine SCNT embryos constructed from Landrace fetal fibroblast cells (FFCs) in vitro and in vivo, but not at higher concentrations. Therefore, we optimized the application of another novel HDACi, Scriptaid, for development of porcine SCNT embryos. We found that treatment with 500 nM Scriptaid significantly enhanced the development SCNT embryos to the blastocyst stage when outbred Landrace FFCs and ear fibroblast cells (EFCs) were used as donors compared to the untreated group. Scriptaid increased the overall cloning efficiency from 0.4% (untreated group) to 1.6% for Landrace FFCs and 0 to 3.7% for Landrace EFCs. Moreover, treatment of SCNT embryos with Scriptaid improved the histone acetylation on Histone H4 at lysine 8 (AcH4K8) in a pattern similar to that of the in vitro fertilized (IVF) embryos.
体细胞核的表观遗传重编程缺陷很可能是通过体细胞核移植（SCNT）产生的所有哺乳动物成功率低的主要原因。已经证明，通过曲古抑菌素A（TSA）等组蛋白去乙酰化酶抑制剂（HDACi）处理以增加组蛋白乙酰化，可显著提高几种物种的SCNT胚胎的发育能力。在此，我们报告，激活后用50 nM TSA处理10小时可提高体外和体内由长白猪胎儿成纤维细胞（FFC）构建的猪SCNT胚胎的发育能力，但更高浓度则无此效果。因此，我们优化了另一种新型HDACi Scriptaid在猪SCNT胚胎发育中的应用。我们发现，与未处理组相比，当使用远交系长白猪FFC和耳成纤维细胞（EFC）作为供体时，用500 nM Scriptaid处理可显著提高SCNT胚胎发育到囊胚阶段的能力。Scriptaid将长白猪FFC的总体克隆效率从0.4%（未处理组）提高到1.6%，将长白猪EFC的克隆效率从0提高到3.7%。此外，用Scriptaid处理SCNT胚胎可改善组蛋白H4赖氨酸8（AcH4K8）上的组蛋白乙酰化，其模式类似于体外受精（IVF）胚胎。
