Scriptaid affects histone acetylation and the expression of development-related genes at different stages of porcine somatic cell nuclear transfer embryo during early development

Abstract

Although the somatic cell nuclear transfer (SCNT) technique has been used extensively for cloning and generating transgenic pigs, the cloning efficiency is still very low. It has been proposed that the low efficiency of this technique is the result of incomplete epigenetic reprogramming and abnormal gene expression during early embryonic development. In this study, we investigate the effect of Scriptaid, a low-toxicity histone deacetylase inhibitor, on the developmental competence of porcine SCNT embryos. We found that treating SCNT embryos with 500 nmol/L Scriptaid for 15 h after activation significantly enhanced the blastocyst formation rate (27.7%) compared with the untreated group (control) (12.2%, P<0.05). Using an immunofluorescence technique to measure the average fluorescence intensity, we also found that treating SCNT embryos with Scriptaid increased the level of histone acetylation on histone H3 at lysine 14 (acH3K14). Furthermore, treating embryos with Scriptaid increased the expression level of three genes that play important roles during embryonic development (Oct4, Klf4 at the blastocyst stage and Nanog at the 4-cell stage). Moreover, the expression level of the apoptosis-related gene Caspase-3 was significantly lower in the Scriptaid-treated SCNT embryos compared with the control SCNT embryos at the 4-cell and blastocyst stages. In conclusion, these results indicate that Scriptaid treatment improves the development and nuclear reprogramming of porcine SCNT embryos.

Improvement of Cloned Embryos Development by Co-Culturing with Parthenotes: A Possible Role of Exosomes/Microvesicles for Embryos Paracrine Communication

“…Oct4, Sox2, c-Myc, Klf4, and Nanog are all considered the pluripotency genes in most mammals, with differences in temporal and spatial expression in different animals (du Puy et al, 2011). These genes are the key players in reprogramming of the NT-derived embryos, and they were reported to be lower in NT embryos if compared to other in vitro-derived embryos (Niwa et al, 2000;Rodríguez et al, 2012;Zhang et al, 2011;Zhou et al, 2013). The full mechanism of network interaction between these genes is still not elucidated in the pig because of their unique expression in this species (du Puy et al, 2011); however, there were some published reports regarding the mutual interaction of two (Rodda et al, 2005) or more pluripotency genes (du Puy et al, 2011).…”

Successful cloning of an adult breeding boar from the novel Chinese Guike No. 1 swine specialized strain

“…Although the exact mechanisms underlying the epigenetic remodeling and reprogramming of somatic cell nuclei in a cytoplasm of both reconstructed oocytes and descendant blastomeres of resultant cloned embryos are still unclear, increasing lines of evidence suggest that improper or incomplete epigenetic modifications of donor nuclear genome, such as DNA methylation and histone acetylation, are closely associated with the low overall efficiency of pig cloning (Zhao et al 2010b ; Whitworth and Prather 2011 ; Lee and Prather 2013 , 2014 ). Up to now, several types of epigenetic drugs, such as non-specific DNA methyltransferase inhibitors (e.g., 5-aza-2′-deoxycytidine) and non-specific histone deacetylase inhibitors (e.g., trichostatin A, scriptaid and oxamflatin), have been used for epigenomic transformation of in vitro cultured nuclear donor cells, in vitro maturing nuclear recipient oocytes and activated nuclear-transferred oocytes, resulting in not only significant enhancement of the in vitro developmental capacity of porcine cloned embryos (Himaki et al 2010 ; Mao et al 2012 , 2015 ; Ning et al 2012 ; Park et al 2012 ; Samiec and Skrzyszowska 2010c , 2011 , 2012a , 2014b ; Xu et al 2013 ; Zhou et al 2013 ; Cong et al 2013 ; Bohrer et al 2014 ; Luo et al 2014 ; Liang et al 2015 ; Samiec et al 2015 ; Whitworth et al 2015 ), but also the improvement in the efficiency of generating viable cloned offspring in pigs (Zhao et al 2009 , 2010a ). Thus, such successful strategy can be used in our future work for producing cloned pigs with a higher efficiency.…”