Effects of trichostatin A on in vitro development and transgene function in somatic cell nuclear transfer embryos derived from transgenic Clawn miniature pig cells

Abstract

The present study was carried out to examine the effects of post-activation treatment of trichostatin A (TSA), a histone deacetylase inhibitor, on in vitro development and transgene function of somatic cell nuclear transfer (SCNT) embryos derived from Clawn miniature pig embryonic fibroblast (PEF) transfected with a bacterial endo-b-galactosidase C gene (removal of the a-galactosyl (Gal) epitope). SCNT embryos were incubated with or without TSA (50 or 100 nmol/L) after activation, cultured in vitro and assessed for cleavage, blastocyst formation and transgene function. The rate of blastocyst formation was significantly higher in SCNT embryos treated with 50 nmol/L TSA than that in control (P < 0.05), whereas the rate of cleavage and cell number of blastocyst did not differ. Following labelling with fluorescein isothiocyanate-labelled BS-I-B4 isolectin, the intensity of fluorescence observed on cell-surface was dramatically reduced in transgenic SCNT blastocyst in comparison with non-transgenic SCNT blastocyst. However, the reduction of a-Gal epitope expression in transgenic SCNT blastocyst was not affected by TSA treatment. The results of this study showed that post-activation treatment with 50 nmol/L TSA is effective to improve in vitro developmental capacity of transgenic SCNT miniature pig embryos without the modification of transgene function.

The Effects of 5-Aza-2′- Deoxycytidine and Trichostatin A on Gene Expression and DNA Methylation Status in Cloned Bovine Blastocysts

“…It has been reported that treatment of donor cells (Enright et al, 2005) or early NT embryos (Tsuji et al, 2009) with 5-aza-dC does not increase the developmental potential of cloned embryos. Several histone deacetylase inhibitors (HDACi) such as trichostatin A, valproic acid, scriptaid, and sodium butyrate, have previously been tested as a means to improve SCNT efficiency in various species, including piglets (Beebe et al, 2009;Das et al, 2010;Li et al, 2008a;Zhang et al, 2007;Zhao et al, 2010), NIH miniature pigs (Himaki et al, 2010;Zhao et al, 2009), miniature pigs (Miyoshi et al, 2010), mice (Costa-Borges et al, 2010;Maalouf et al, 2009;Rybouchkin et al, 2006;Van Thuan et al, 2009), rabbits (Shi et al, 2008;Yang et al, 2007a), and cattle (Ding et al, 2008; Iager et al, 2008;Wee et al, 2007). To date, the effect of the HDACi on the developmental competence has been thoroughly studied, whereas there are few investigations that examined how HDACi enhance the epigenetic remodeling ability of somatic cell nuclei in SCNT embryos.…”

“…To improve cloning efficiency, several epigenetic remodeling drugs such as DNA methylation inhibitor 5-aza-2 0 -deoxycytidine (5-aza-dC) (Ding et al, 2008;Enright et al, 2005;Tsuji et al, 2009), histone deacetylase inhibitor (HDACi), trichostatin A (TSA) (Beebe et al, 2009;CostaBorges et al, 2010;Ding et al, 2008;Himaki et al, 2010;Iager et al, 2008;Kishigami et al, 2006;Li et al, 2008a;Maalouf et al, 2009;Meng et al, 2009;Rybouchkin et al, 2006;Shi et al, 2008;Zhang et al, 2007), valproic acid (Costa-Borges et al, 2010;Miyoshi et al, 2010), scriptaid (Van Thuan et al, 2009;Zhao et al, 2009Zhao et al, , 2010, and sodium butyrate (Das et al, 2010;Shi et al, 2003;Yang et al, 2007a) have recently been tested to improve the developmental competence of cloned embryos. For instance, two independent studies showed that TSA treatment improves the ability of mouse SCNT oocytes to develop into blastocyst and progress to full term, thereby significantly improving mouse cloning efficiency (Kishigami et al, 2006;Rybouchkin et al, 2006).…”

The HDAC Inhibitor LAQ824 Enhances Epigenetic Reprogramming and In Vitro Development of Porcine SCNT Embryos

“…Recently, there were several reports that HDACi treatment positively regulates histone acetylation levels, epigenetic reprogramming, gene expression, full term embryo development and DNA methylation levels in a manner similar to these events taking place in in vitro fertilized embryos. The development of porcine SCNT embryos was improved by treatment with HDACi, such as trichostatin A (TSA) [10, 11], valproic acid (VPA) [12], scriptaid [13, 14], oxamflatin [15], sodium butyrate (NaBu) [16], LBH589 [17], CUDC-101 [18], m-carboxycinnamic acid bishydroxamide (CBHA) [3], PXD101 [19], suberoylanilide hydroxamic acid (SAHA) [20] and PCI-24781 [21]. …”

Effects of Histone Acetylation Status on the Early Development ofIn VitroPorcine Transgenic Cloned Embryos

“…However, abnormal transgene expression level and low cloning efficiency are major limitations to producing transgenic animals. Recently, investigations have focused on the ability of HDAC inhibitors to improve SCNT efficiency, although the effects of HDAC inhibitors varied with treatment duration, concentration, species (Bui et al, 2007;Himaki et al, 2010;Lee et al, 2011;Li et al, 2008b;Shi et al, 2008;Zhao et al, 2009), and the type used. Results of many experiments have indicated that acetylation status played a very important role in the process of reprogramming and affected the development of SCNT embryos (Dai et al, 2010;Das et al, 2010;Ding et al, 2008;Li et al, 2008a).…”