Reduced levels of intracellular reactive oxygen species and apoptotic status are not correlated with increases in cryotolerance of bovine embryos produced in vitro in the presence of antioxidants
Nathália A. S. Rocha-Frigoni A , Beatriz C. S. Leão A , Ériklis Nogueira B , Mônica F. Accorsi A and Gisele Z. Mingoti A C
+ Author Affiliations
- Author Affiliations
A Department of Animal Health School of Veterinary Medicine, UNESP–Universidade Estadual Paulista, Araçatuba, SP 16050-680, Brazil.
B Brazilian Agricultural Research Corporation, EMBRAPA Pantanal, Corumbá, MS 79320-900, Brazil.
C Corresponding author. Email: gmingoti@fmva.unesp.br
Reproduction, Fertility and Development 26(6) 797-805 https://doi.org/10.1071/RD12354
Submitted: 2 November 2012 Accepted: 14 May 2013 Published: 6 June 2013
Abstract
The effects of intracellular (cysteine and β-mercaptoethanol) and extracellular (catalase) antioxidant supplementation at different times during in vitro production (IVM and/or in vitro culture (IVC)) on bovine embryo development, intracellular reactive oxygen species (ROS) levels, apoptosis and re-expansion rates after a vitrification–thawing process were examined. Blastocyst frequencies were not affected by either antioxidant supplementation (40.5%–56.4%) or the timing of supplementation (41.7%–55.4%) compared with control (48.7%; P > 0.05). Similarly, antioxidants and the moment of supplementation did not affect (P > 0.05) the total number of blastomeres (86.2–90.5 and 84.4–90.5, respectively) compared with control (85.7). However, the percentage of apoptotic cells was reduced (P < 0.05) in groups supplemented during IVM (1.7%), IVC (2.0%) or both (1.8%) compared with control (4.3%). Intracellular ROS levels measured in Day 7 blastocysts were reduced (P < 0.05) in all groups (0.60–0.78), with the exception of the group supplemented with β-mercaptoethanol during IVC (0.88), which did not differ (P > 0.05) from that in the control group (1.00). Re-expansion rates were not affected (P > 0.05) by the treatments (50.0%–93.0%). In conclusion, antioxidant supplementation during IVM and/or IVC reduces intracellular ROS and the rate of apoptosis; however, supplementation does not increase embryonic development and survival after vitrification.
Additional keywords: oxidative stress, redox status.
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