Cotreatment with RepSox and LBH589 improves the in vitro developmental competence of porcine somatic cell nuclear transfer embryos.
Cotreatment with RepSox and LBH589 improves the in vitro developmental competence of porcine somatic cell nuclear transfer embryos.
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DOI：
10.1071/rd17543
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发表时间：
2018-10
期刊：
影响因子：
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通讯作者：
Zhao-Bo Luo;Long Jin;Q. Guo;Junxia Wang;Xiao-Xu Xing;Meifu Xuan;Qi-Rong Luo;Guang-Lei Zhang;Xi-jun Yin;Jin-Dan Kang
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文献类型：
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作者：
Zhao-Bo Luo;Long Jin;Q. Guo;Junxia Wang;Xiao-Xu Xing;Meifu Xuan;Qi-Rong Luo;Guang-Lei Zhang;Xi-jun Yin;Jin-Dan Kang
越来越多的证据表明，供体细胞核异常的表观遗传重编程和多能性较低，会导致克隆胚胎发育异常，这也是哺乳动物体细胞核移植（SCNT）效率低下的根本原因。本研究表明，单独使用小分子RepSox处理可上调猪体细胞核移植胚胎中多能性相关基因的表达。使用组蛋白去乙酰化酶抑制剂LBH589处理能显著提高囊胚形成率，而RepSox处理则无此效果。与未处理的胚胎相比，用12.5μM的RepSox和50nM的LBH589共同处理（RepSox + LBH589）24小时，囊胚形成率显著提高（分别为26.9%和8.5%；P < 0.05）。此外，在4细胞期和囊胚期，与对照组相比，RepSox + LBH589处理组中多能性相关基因八聚体结合转录因子4（NANOG）和Y染色体性别决定区框蛋白2（SOX2）的表达显著增加。特别是在囊胚期，RepSox + LBH589组中NANOG的表达量比对照组高135倍。此外，RepSox + LBH589改善了表观遗传重编程。总之，RepSox + LBH589可增加发育重要基因的表达，优化表观遗传重编程，并改善猪体细胞核移植胚胎的体外发育。
Accumulating evidence suggests that aberrant epigenetic reprogramming and low pluripotency of donor nuclei lead to abnormal development of cloned embryos and underlie the inefficiency of mammalian somatic cell nuclear transfer (SCNT). The present study demonstrates that treatment with the small molecule RepSox alone upregulates the expression of pluripotency-related genes in porcine SCNT embryos. Treatment with the histone deacetylase inhibitor LBH589 significantly increased the blastocyst formation rate, whereas treatment with RepSox did not. Cotreatment with 12.5μM RepSox and 50nM LBH589 (RepSox+LBH589) for 24h significantly increased the blastocyst formation rate compared with that of untreated embryos (26.9% vs 8.5% respectively; P<0.05). Furthermore, the expression of pluripotency-related genes octamer-binding transcription factor 4 (NANOG) and SRY (sex determining region Y)-box 2 (SOX2) were found to significantly increased in the RepSox+LBH589 compared with control group at both the 4-cell and blastocyst stages. In particular, the expression of NANOG was 135-fold higher at the blastocyst stage in the RepSox+LBH589 group. Moreover, RepSox+LBH589 improved epigenetic reprogramming. In summary, RepSox+LBH589 increases the expression of developmentally important genes, optimises epigenetic reprogramming and improves the invitro development of porcine SCNT embryos.