Simulating the effects of H2O2 and melatonin treatments on the oxidation and reduction state of sperm on two outcomes: DNA uptake in sperm and apoptosis of embryonic cells

Abstract

Abstract Improving the transfer of foreign genes into recalcitrant cells, as in the production of transgenic animal models, is a challenge. Sperm-mediated gene transfer (SMGT) is a cost-effective and rapid method for exogenous DNA transferring into egg by sperm during fertilization. This method can be improved using chemical and natural additives. The objective of this study was to examine how sperm function, DNA uptake, embryonic development, and apoptosis rates are affected by different levels of ROS under varying oxidant and antioxidant conditions in SMGT method. Sperm isolated from mice with and without plasmid were treated with varying concentrations of H2O2 and melatonin. Under oxidation conditions, sperm performance was significantly reduced compared to the melatonin treatment. Embryos from the oxidation group with 100 µM H2O2 treatment showed higher uptake of exogenous DNA, and the proportion of GFP-positive blastocysts was significantly higher than those from higher concentrations. The data demonstrate that oxidative conditions in sperm are associated with increased absorption of exogenous DNA, potentially due to an increase in the mitochondria ability resulting from reactive oxygen species increase and sperm membrane instability. The importance of antioxidants lies in their ability to safeguard DNA against oxidative stress, enhance absorption efficacy, and bolster the stability of the sperm membrane. The study showed that antioxidants reduced the risk of negative effects on sperm quality and function.