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

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.