Abstract
Melatonin has antioxidant and scavenger effects in the cellular antioxidant system. This research investigated the protective effects and underlying mechanisms of melatonin action in porcine somatic cell nuclear transfer (SCNT) embryos. The results suggested that the developmental competence of porcine SCNT embryos was considerably enhanced after melatonin treatment. In addition, melatonin attenuated the increase in reactive oxygen species levels induced by oxidative stress, the decrease in glutathione levels, and the mitochondrial dysfunction. Importantly, melatonin inhibited phospho-histone H2A.X (γH2A.X) expression and comet tail formation, suggesting that γH2A.X prevents oxidative stress-induced DNA damage. The expression of genes involved in homologous recombination and non-homologous end-joining pathways for the repair of double-stranded breaks (DSB) was reduced upon melatonin treatment in porcine SCNT embryos at day 5 of development under oxidative stress condition. These results indicated that melatonin promoted porcine SCNT embryo development by preventing oxidative stress-induced DNA damage via quenching of free radical formation. Our results revealed a previously unrecognized regulatory effect of melatonin in response to oxidative stress and DNA damage. This evidence provides a novel mechanism for the improvement in SCNT embryo development associated with exposure to melatonin.
Highlights
Somatic cell nuclear transfer (SCNT) is an extraordinary and important technology for generating transgenic animals and preserving species[1,2,3]
The numbers of embryos examined in each group are shown in the bars
Data are expressed as the mean ± standard deviation (SD) from at least three separate experiments. *p < 0.05; **p < 0.01
Summary
Somatic cell nuclear transfer (SCNT) is an extraordinary and important technology for generating transgenic animals and preserving species[1,2,3]. DNA methyltransferase inhibitors such as BIX-01294 and 5′-azacytidine (5′-azaC), and histone deacetylase inhibitors such as trichostatin A, scriptaid, and valproic acid have greatly improved the cloning efficiency in several species[9,10,11,12,13]. Another major problem during the production of SCNT embryos is the variation in culture conditions. The HDACi scriptaid effectively enhances DNA damage repair and improves somatic cell reprogramming by increasing the ability of SCNT embryos to develop in vitro[30]. Su et al15. demonstrated that supplementation with exogenous melatonin reduced apoptosis and levels of ROS in bovine SCNT embryos, which improved overall cloning efficiency
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