Oxidative and nitrosative stress in frozen-thawed pig spermatozoa. II: Effect of the addition of saccharides to freezing medium on sperm function |
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| Affiliation: | 1. Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80526, USA;2. Acuity Genetics, Carlyle, IL 62231, USA;3. Membrane Protective Technologies Inc., Fort Collins, CO 80524, USA;1. South-East Zoo Alliance for Reproduction & Conservation, 581705 White Oak Road, Yulee, FL, 32097, USA;2. School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington, Loughborough, LE12 5RD, UK;3. Academic Department of Reproductive and Developmental Medicine, University of Sheffield, Level 4, Jessop Wing, Tree Root Walk, Sheffield, S10 2SF, UK;4. Department of Biomedical Sciences, Colorado State University, Fort Collins, CO, 80521, USA;5. School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD, UK;1. Centro de Pruebas de Porcino, Área de Investigación Ganadera, Subdirección de Investigación y Tecnología, Instituto Tecnológico Agrario, Consejería de Agricultura y Ganadería, Junta de Castilla y León, Ctra Riaza-Toro s/n, 40353 Hontalbilla, Segovia, Spain;2. Centro de Investigación y Tecnología Animal – Instituto Valenciano de Investigaciones Agrarias (CITA-IVIA), 12400 Segorbe, Castellón, Spain |
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| Abstract: | Saccharides have bioprotective properties, with a high capacity to preserve biological proteins and membranes during sperm cryopreservation. The aim of this study was to evaluate how replacing the lactose of cryopreservation media by sucrose (SUC) or trehalose (TRE) at concentrations of 0.2 M (SUC-1 and TRE-1) and 0.25 M (SUC-2 and TRE-2) affects frozen/thawed pig spermatozoa. The media used were composed of medium A (saccharide/egg yolk) and B (saccharide/egg yolk/glycerol), their osmolality being determined prior to freezing. Cell viability, membrane lipid disorder, acrosome integrity, mitochondrial membrane potential (MMP), lipid peroxidation, thiol group oxidation, total reactive oxygen species (ROS), peroxynitrite and superoxide anion (O2●-) were determined through flow cytometry; total motility (TM), progressive motility (PM) and kinetic parameters motility were determined immediately after thawing (T0) and again 30 (T30) and 60 (T60) minutes later. The SUC-2 and TRE-2 groups maintained viability significantly and presented fewer lipid membrane disorders, respectively, both with a significant increase in MMP. The production of O2●- and peroxynitrite was lower in the TRE-2 groups compared to the control (P < 0.05). Total motility at T0 was greater in the TRE-2 group (P < 0.05). Sperm kinetics was not affected by the treatment. The use of saccharides SUC and TRE at a concentration of 0.25 M improves sperm quality, so that both non-penetrating cryoprotectants can be utilized in pig sperm freezing media. |
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| Keywords: | Trehalose Sucrose Cryopreservation Spermatozoa Pig Reproduction Oxidative stress Nitrosative stress |
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