Cotreatment with RepSox and LBH589 improves the in vitro developmental competence of porcine somatic cell nuclear transfer embryos
Zhao-Bo Luo A , Long Jin A , Qing Guo A , Jun-Xia Wang A , Xiao-Xu Xing A , Mei-Fu Xuan A , Qi-Rong Luo A , Guang-Lei Zhang A , Xi-Jun Yin A B and Jin-Dan Kang A B
+ Author Affiliations
- Author Affiliations
A Jilin Provincial Key Laboratory of Transgenic Animal and Embryo Engineering. Yanbian University, No. 977 Gongyuan Street, Yanji City, Jilin Pro 133002, China.
B Corresponding authors. Email: kangjindan@hotmail.com; yinxj33@msn.com
Reproduction, Fertility and Development 30(10) 1342-1351 https://doi.org/10.1071/RD17543
Submitted: 11 October 2017 Accepted: 22 March 2018 Published: 24 April 2018
Abstract
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 12.5 μM RepSox and 50 nM LBH589 (RepSox + LBH589) for 24 h 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 in vitro development of porcine SCNT embryos.
Additional keywords: histone deacetylase inhibitor, NANOG.
References
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