Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning, China.
Theriogenology. 2013 Nov;80(8):878-86. doi: 10.1016/j.theriogenology.2013.07.013. Epub 2013 Sep 3.
It has been reported that buffalo (Bubalus bubalis) embryos reconstructed by somatic cell nucleus transfer (SCNT) can develop to the full term of gestation and result in newborn calves. However, the developmental competence of reconstructed embryos is still low. Recently, it has been reported that treating donor cells or embryos with trichostatin A (TSA) can increase the cloning efficiency in some species. Thus, the present study was undertaken to improve the development of buffalo SCNT embryos by treatment of donor cells (buffalo fetal fibroblasts) with TSA and explore the relation between histone acetylation status of donor cells and developmental competence of SCNT embryos. Treatment of donor cells with either 0.15 or 0.3 μM TSA for 48 hours resulted in a significant increase in the cleavage rate and blastocyst yield of SCNT embryos (P < 0.05). Meanwhile, the expression level of HDAC1 in donor cells was also decreased (0.4-0.6 fold, P < 0.05) by TSA treatment, although the expression level of HAT1 was not affected. Further measurement of the epigenetic maker AcH4K8 in buffalo IVF and SCNT embryos at the eight-cell stage revealed that the spatial distribution of acH4K8 staining in SCNT embryos was different from the IVF embryos. Treatment of donor cells with TSA resulted in an increase in the AcH4K8 level of SCNT embryos and similar to fertilized counterparts. These results suggest that treatment of donor cells with TSA can facilitate their nucleus reprogramming by affecting the acetylated status of H4K8 and improving the in vitro development of buffalo SCNT embryos. The AcH4K8 status at the eight-cell stage can be used as an epigenetic marker for predicting the SCNT efficiency in buffalos.
据报道，通过体细胞核移植（SCNT）重构的水牛胚胎可以发育到足月并产生新生牛犊。然而，重构胚胎的发育能力仍然较低。最近，有报道称，用曲古抑菌素 A（TSA）处理供体细胞或胚胎可以提高一些物种的克隆效率。因此，本研究通过用 TSA 处理供体细胞（水牛胎儿成纤维细胞）来提高水牛 SCNT 胚胎的发育，并探讨供体细胞组蛋白乙酰化状态与 SCNT 胚胎发育能力之间的关系。用 0.15 或 0.3 μM TSA 处理供体细胞 48 小时，可显著提高 SCNT 胚胎的卵裂率和囊胚率（P < 0.05）。同时，TSA 处理也降低了供体细胞中 HDAC1 的表达水平（0.4-0.6 倍，P < 0.05），而 HAT1 的表达水平不受影响。进一步测量体外受精和 SCNT 胚胎在 8 细胞阶段的组蛋白乙酰化标记物 AcH4K8 表明，SCNT 胚胎中 AcH4K8 染色的空间分布与体外受精胚胎不同。用 TSA 处理供体细胞可增加 SCNT 胚胎的 AcH4K8 水平，并与受精胚胎相似。这些结果表明，用 TSA 处理供体细胞可以通过影响 H4K8 的乙酰化状态来促进核重编程，从而提高水牛 SCNT 胚胎的体外发育。8 细胞阶段的 AcH4K8 状态可用作预测水牛 SCNT 效率的表观遗传标记。
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