Impact of Seminal Plasma Antioxidants on DNA Fragmentation and Lipid Peroxidation of Frozen-Thawed Horse Sperm

Jaime Catalán; Iván Yánez-Ortiz; Marc Torres-Garrido; Jordi Ribas-Maynou; Marc Llavanera; Isabel Barranco; Marc Yeste; Jordi Miró

doi:10.3390/antiox13030322

Impact of Seminal Plasma Antioxidants on DNA Fragmentation and Lipid Peroxidation of Frozen-Thawed Horse Sperm

Antioxidants (Basel). 2024 Mar 6;13(3):322. doi: 10.3390/antiox13030322.

Authors

Jaime Catalán^{1

2

3}, Iván Yánez-Ortiz^{1

2

3

4}, Marc Torres-Garrido^{1

2}, Jordi Ribas-Maynou^{1

2}, Marc Llavanera^{1

2}, Isabel Barranco⁵, Marc Yeste^{1

2

6}, Jordi Miró³

Affiliations

¹ Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, ES-17003 Girona, Spain.
² Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, ES-17003 Girona, Spain.
³ Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain.
⁴ School of Veterinary Medicine, Faculty of Medical, Health and Life Sciences, International University of Ecuador, Quito 170411, Ecuador.
⁵ Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Murcia, ES-30100 Murcia, Spain.
⁶ Catalan Institution for Research and Advanced Studies (ICREA), ES-08010 Barcelona, Spain.

Abstract

Cryopreservation is a stressful process for sperm, as it is associated with an increased production of reactive oxygen species (ROS). Elevated ROS levels, which create an imbalance with antioxidant capacity, may result in membrane lipid peroxidation (LPO), protein damage and DNA fragmentation. This study aimed to determine whether the membrane LPO and DNA fragmentation of frozen-thawed horse sperm relies upon antioxidant activity, including enzymes (superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT) and paraoxonase type 1 (PON1)); non-enzymatic antioxidant capacity (Trolox-equivalent antioxidant capacity (TEAC), plasma ferric reducing antioxidant capacity (FRAP) and cupric reducing antioxidant capacity (CUPRAC)); and the oxidative stress index (OSI) of their seminal plasma (SP). Based on total motility and plasma membrane integrity (SYBR14⁺/PI^-) after thawing, ejaculates were hierarchically (p < 0.001) clustered into two groups of good- (GFEs) and poor-(PFEs) freezability ejaculates. LPO and DNA fragmentation (global DNA breaks) were higher (p < 0.05) in the PFE group than in the GFE group, with LPO and DNA fragmentation (global DNA breaks) after thawing showing a positive relationship (p < 0.05) with SP OSI levels and ROS production. In addition, sperm motility and membrane integrity after thawing were negatively (p < 0.05) correlated with the activity levels of SP antioxidants (PON1 and TEAC). The present results indicate that LPO and DNA fragmentation in frozen-thawed horse sperm vary between ejaculates. These differences could result from variations in the activity of antioxidants (PON1 and TEAC) and the balance between the oxidant and antioxidant components present in the SP.

Keywords: DNA fragmentation; antioxidant; cryopreservation; horse; lipid peroxidation; semen; seminal plasma; sperm.

Grants and funding

J.C. was funded by the Ministry of Universities (Spain) and by the European Union Next Generation EU Funds (Margarita Salas Scheme), and I.B. was funded by the Spanish Ministry of Science and Innovation (MCIN/AEI/10.13039/501100011033) and the European Union NextgenerationEU/PRTR (Ramón y Cajal Scheme: RYC2021-034546-I). The authors also acknowledge the support from the Ministry of Science and Innovation, Spain (Grant: PID2020-113320RB-I00); the Catalan Agency for the Management of University and Research Grants, Regional Government of Catalonia, Spain (Grant: 2017-SGR-1229 and 2021-SGR-00900); and the Catalan Institution for Research and Advanced Studies, Spain (ICREA).