Comparison between the Effects of Valproic Acid and Trichostatin A on the In Vitro Development, Blastocyst Quality, and Full-Term Development of Mouse Somatic Cell Nuclear Transfer Embryos
Comparison between the Effects of Valproic Acid and Trichostatin A on the In Vitro Development, Blastocyst Quality, and Full-Term Development of Mouse Somatic Cell Nuclear Transfer Embryos
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DOI：
10.1089/cell.2009.0108
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发表时间：
2010-08-01
影响因子：
1.6
通讯作者：
Ibanez, Elena
中科院分区：
文献类型：
--
作者：
Costa-Borges, Nuno;Santalo, Josep;Ibanez, Elena
体细胞核移植（SCNT）后分化核重编程为全能性胚胎状态是不有效的。先前对杂交B6D2F1小鼠株的研究表明，用组蛋白去乙酰化酶抑制剂trichostatin a （TSA）短暂处理SCNT胚胎可显著提高克隆胚胎在体外发育和足月的潜力。在这里，我们比较了两种不同的SCNT方案与TSA，并首次探讨了另一种HDACi丙戊酸（VPA）对小鼠B6CBAF1克隆胚胎的体外发育、囊胚质量和足月发育的影响。5 nM TSA处理的SCNT胚胎在激活期间和激活后的囊胚发育率（31.8%），或100 nM TSA或2 mM VPA处理的SCNT胚胎在激活前和激活期间的囊胚发育率（分别为34.5%和38.3%）明显优于未处理的SCNT胚胎（22.9-25.1%）。与未处理的胚胎相比，hdaci处理的胚胎体外发育率的提高与组蛋白H3赖氨酸14乙酰化水平的提高和囊胚质量的改善有关（通过总细胞和ICM细胞数量的增加来判断）（增加30-35%）。用TSA或VPA处理SCNT胚胎也可以观察到存活的克隆小鼠，而未经处理的SCNT胚胎不能产生存活的克隆小鼠。总之，我们首次证明VPA可以改善B6CBAF1 SCNT胚胎的体外和足月发育，其水平与TSA相似。我们的发现可能为提高其他小鼠品系或物种的克隆效率开辟新的机会。
Reprogramming of differentiated nuclei into a totipotent embryonic state following somatic cell nuclear transfer (SCNT) is not efficient. Previous studies in the hybrid B6D2F1 mouse strain revealed that a transient treatment of the SCNT embryos with the histone deacetylase inhibitor (HDACi) trichostatin A (TSA) significantly enhance the potential of the cloned embryos to develop in vitro and to term. Here, we compare two different SCNT protocols with TSA and explore, for the first time, the effect of another HDACi, valproic acid (VPA), on the in vitro development, blastocyst quality, and full-term development of mouse B6CBAF1 cloned embryos. Rates of blastocyst development in SCNT embryos treated with either 5 nM TSA during and after activation (31.8%) or with 100 nM TSA or 2 mM VPA before and during activation (34.5 and 38.3%, respectively) were clearly superior to those of nontreated SCNT embryos (22.9-25.1%). These increased in vitro development rates of the HDACi-treated embryos were correlated with an increased level of histone H3 lysine 14 acetylation and an improved blastocyst quality, as judged by the increased number of total and ICM cells in comparison to the nontreated embryos (30-35% increase). Treatment of SCNT embryos with TSA or VPA also allowed the obtention of viable cloned mice, whereas none could be produced from untreated SCNT embryos. In conclusion, we have demonstrated for the first time that VPA can improve the in vitro and full-term development of B6CBAF1 SCNT embryos, at a similar level as TSA. Our findings may open new opportunities to improve cloning efficiencies in other mouse strains or species.