Lee Min-Jung, Kim Se-Woong, Lee Hyun-Gi, Im Gi-Sun, Yang Byoung-Chul, Kim Nam-Hyung, Kim Dong-Hoon
National Institute of Animal Science, Suwon 441-706, Korea.
J Reprod Dev. 2011 Feb;57(1):34-42. doi: 10.1262/jrd.10-012a. Epub 2010 Sep 10.
We studied the effects of trichostatin A (TSA), a histone deacetylase inhibitor, on the development of bovine somatic cell nuclear transfer (SCNT) embryos by investigating (1) the optimal concentration and treatment time of TSA for development of bovine SCNT embryos, (2) the status of histone acetylation in TSA-treated and control SCNT embryos and (3) the expression of histone acetylation- and deacetylation-related genes in TSA-treated and control SCNT embryos. We observed that 50 nM TSA-treatment for 20 h following fusion resulted in more efficient in vitro development of bovine SCNT embryos to the blastocyst stage. In regard to histone H4K5 acetylation, half of the control SCNT embryos faintly displayed histone H4K5 signals 30 min after electrofusion, while most of the TSA-treated SCNT embryos displayed histone H4K5 signals within 30 min after electrofusion. Furthermore, the expressions of HDAC1 and HDAC2 in the blastocysts were significantly lower (P<0.05) in the TSA-treated SCNT than in the control SCNT. However, the expression of GCN5 and HAT1 did not differ between the TSA-treated and control SCNT. In conclusion, we demonstrated that TSA-treatment after SCNT in bovine embryos can dramatically improve the practical applications of current cloning techniques.
我们通过研究（1）曲古抑菌素A（TSA，一种组蛋白去乙酰化酶抑制剂）对牛体细胞核移植（SCNT）胚胎发育的最佳浓度和处理时间，（2）经TSA处理和未经处理的SCNT胚胎中组蛋白乙酰化状态，以及（3）经TSA处理和未经处理的SCNT胚胎中组蛋白乙酰化和去乙酰化相关基因的表达，来研究TSA对牛SCNT胚胎发育的影响。我们观察到，融合后用50 nM TSA处理20小时可使牛SCNT胚胎在体外更有效地发育至囊胚阶段。关于组蛋白H4K5乙酰化，一半的对照SCNT胚胎在电融合后30分钟微弱显示组蛋白H4K5信号，而大多数经TSA处理的SCNT胚胎在电融合后30分钟内显示组蛋白H4K5信号。此外，经TSA处理的SCNT胚胎囊胚中HDAC1和HDAC2的表达显著低于对照SCNT胚胎（P<0.05）。然而，经TSA处理和对照的SCNT胚胎中GCN5和HAT1的表达没有差异。总之，我们证明了牛胚胎SCNT后进行TSA处理可显著改善当前克隆技术的实际应用。
