Abstract

Oxidative stress is associated with impaired sperm quality after thawing. Since mitochondria are the main source of reactive oxygen species (ROS) in sperm, the aim of this study was to investigate the effects of targeted mitochondrial antioxidant mitoquinone (MitoQ) and non-targeted mitochondrial antioxidant pentoxifylline (PTX) during cooling and cryopreservation of rooster sperm. Sperm samples were collected from 15 roosters aged 28 wk and diluted with Beltsville extender. After dilution and addition of treatments (50, 100, and 200 pMol MitoQ and 0.5, 0.75, and 1 μM PTX), samples were cooled for 2 h to 4°C and they were first analyzed at this stage and were frozen and re-evaluated after thawing. After the freezing and thawing, level of 100 pMol MitoQ significantly increased total motility (TM), progressive motility (PGM), curvilinear velocity (VCL), membrane integrity, viability, total antioxidant capacity (TAC) and the glutathione peroxidase (GPx), as well as the level of 50 pMol significantly increased TM, PGM, average path velocity (VAP), straight-line velocity (VSL), membrane integrity, viability, and mitochondrial activity. Moreover, these 2 levels (50 and 100 PMol) decreased malondialdehyde and sperm with abnormal morphology. Addition of 0.75 μM PTX also increased total motility compared to the control group and levels of 0.5 and 0.75 μM decreased sperm with abnormal morphology. It could be concluded the addition of MitoQ and PTX can be useful for sperm cryopreservation industry and reduce the harmful effects of freeze-thawing.

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