Effect of antifreeze protein I in the freezing solution on in vivo-derived sheep embryos

• Published in: Research in veterinary science 2024-03, Vol.168, p.105132-105132, Article 105132
• Main Authors: Correia, Lucas F.L., Leal, Gabriela R., Brandão, Felipe Z., Batista, Ribrio I.T.P., Souza-Fabjan, Joanna M.G.
• Format: Article
• Language: English
• Subjects: alpha-Fetoproteins; Animals; Antifreeze Proteins; apoptosis; Blastocyst; cryopreservation; Cryopreservation - veterinary; Cryoprotective Agents - pharmacology; Cryoprotector; Cryosurvival; Freezing; glutathione; homeostasis; Metabolism; mitochondria; oxidative stress; Proto-Oncogene Proteins c-bcl-2; reactive oxygen species; Sheep; Slow freezing; survival rate; veterinary medicine; viability
• ISSN: 0034-5288; 1532-2661
• DOI: 10.1016/j.rvsc.2023.105132

This study evaluated the effects of different antifreeze protein type I (AFP I) concentrations added to a slow freezing solution in sheep in vivo-derived embryos. Good-quality embryos were allocated into: AFP-free (CONT); 0.1 μg/mL of AFP I (AFP0.1); or 0.5 μg/mL of AFP I (AFP0.5). After thawing, embryos were in vitro cultured (IVC) for 48 h. At 24 h and 48 h of IVC, dead cells and apoptosis, mitochondrial activity, intracellular reactive oxygen species (ROS), and glutathione (GSH) evaluations were performed. At 24 h, evaluated embryos were submitted to RT-qPCR for metabolism (SIRT2, PRDX1, OCT4, CDX2) and quality (AQP3, CDH1, HSP70, BAX, BCL2) genes. The in vitro survival rate was 56% (22/39) for CONT, 60% (32/53) for AFP0.1, and 53% (23/43) for AFP0.5 (p > 0.05). A tendency (p = 0.09) for a higher blastocyst hatching rate was noted in AFP0.1 (62%) compared to AFP0.5 (33%), and both groups were similar to CONT (50%). An increased (p  0.05) were observed in oxidative stress homeostasis and viability between treatments. A downregulation (p