Vitrification of bovine oocytes can impair subsequent embryo development mostly due to elevated oxidative stress. This study was aimed at examining whether glutathione, a known antioxidant, can improve further embryo development when added to devitrified oocytes for a short recovery period. Bovine in vitro matured oocytes were vitrified using an ultra-rapid cooling technique on electron microscopy grids. Following warming, the oocytes were incubated in the recovery medium containing glutathione (0, 1.5, or 5 mmol L-1) for 3 h (post-warm recovery). Afterwards, the oocytes were lysed for measuring the total antioxidant capacity (TAC), activity of peroxidase, catalase and glutathione reductase, and ROS formation. The impact of vitrification on mitochondrial and lysosomal activities was also examined. Since glutathione, added at 5 mmol L-1, significantly increased the TAC of warmed oocytes, in the next set of experiments this dose was applied for post-warm recovery of oocytes used for IVF. Glutathione in the recovery culture did not change the total blastocyst rate, while increased the proportion of faster developing blastocysts (Day 6-7), reduced the apoptotic cell ratio and reversed the harmful impact of vitrification on the actin cytoskeleton. These results suggest that even a short recovery culture with antioxidant(s) can improve the development of bovine devitrified oocytes.

Agricultural and Food Centre Research Institute for Animal Production Nitra Hlohovecká 2 95141 Lužianky Slovakia

Department of Biochemistry Faculty of Science Palacký University Olomouc Šlechtitelů 27 77900 Olomouc Czech Republic

Department of Botany and Genetics Constantine the Philosopher University Nitra Tr A Hlinku 1 94974 Nitra Slovakia

Department of Zoology Faculty of Science Palacký University Olomouc 17 Listopadu 50 77900 Olomouc Czech Republic

Institute of Biotechnology Faculty of Biotechnology and Food Science Slovak University of Agriculture in Nitra Tr A Hlinku 2 94976 Nitra Slovakia

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Reactive oxygen and nitrogen species: multifaceted regulators of ovarian activity†

Astaxanthin Added during Post-Warm Recovery Mitigated Oxidative Stress in Bovine Vitrified Oocytes and Improved Quality of Resulting Blastocysts