The effect of the mitochondrial-directed antioxidant Mito-TEMPO on the quality of rooster sperm in the cryopreservation and lyophilization cycle

Abstract

In reproductive technologies applied in animal husbandry, using lyophilized semen is only possible with ICSI technology, due to complete loss sperm motility. However, the anatomical structure’s peculiarities and development of avian oocyte not allows use this technology. Strategies aimed at preserving kinetic apparatus of lyophilized sperm are the most relevant for birds. Increasing mitochondrial activity of cells due to its connection with energy metabolism and sperm motility may be one of the ways to solve this problem. However, similar studies on freeze-dried bird semen haven’t been conducted on this moment. The present study assessed effect of mitochondria-targeted antioxidant Mito-TEMPO on viability, mitochondria’s membrane potential and generation of intracellular hydrogen peroxide by rooster spermatozoa in a cryopreservation and lyophilization cycle. Proportion of cells with intact plasma membrane in lyophilized semen was higher using extenders containing an antioxidant. Addition of Mito-TEMPO at concentration of 40 µM to diluents T-20 and T-30 for the purpose of cryopreservation of semen with subsequent freezedrying allowed to increase proportion of cells with intact plasma membrane in lyophilized semen in comparison to control diluents (6.12 ± 0.11 % versus 10.19±0.46% and 13.19±1.30% versus 15.23±1.13%, respectively, p˂0.05). Effective concentration of Mito-TEMPO in diluents intended only for semen cryopreservation was significantly lower - 5 µM. Proportion of cells with intact plasma membrane was 47,97±0,64% versus 55,85±1,69% and 47,25±2,97% versus 63,21±1,78%, respectively, p˂0, 05. Dose-dependent effect of trehalose in cryoprotective diluents has been proven.