VARIATION OF SPERM MOTILITY AND MITOCONDRIAL POTENTIAL PARAMETERS DUE TO THE ANTIOXIDANT EFFECT OF TROLOX ON BOAR SEMEN REFRIGERATION

CAMPORINO, A.; CÓRDOBA, M.

CABA

Jornada; XXI Jornadas Anuales de la SAB 2020; 2021

Sociedad Argentina de Biología

Gamete cryopreservation is a fundamental biotechnological strategy in the productive field. However, in swine production, it is not regularly used due to the high concentration of unsaturated fatty acids makes boar sperm more susceptible to peroxidation produced by reactive oxygen species generated during their storage at low temperatures. Our aim was to study the effect of the addition of the synthetic compound derived from alpha-tocopherol called ?Trolox? to Modena modified diluent in porcine semen by means of functional sperm tests: evaluation of sperm motility, pre-capacitation status, sperm viability, membrane integrity, and mitochondrial membrane potential. We worked with a pool of semen samples that were separated into two equal aliquots, only one of them treated with Trolox, and both stored at 17°C. Both samples were evaluated on days 0 (fresh semen), 1, 2 and 5 of refrigeration. Subsequently, motility changes were studied using the Integrated Sperm Analysis System (software version 1.2, PROISER). Pre-capacitation was evaluated by the chlorotetracycline epifluorescent technique and viability by trypan blue staining. The functional integrity of the plasma membrane was evaluated through the Hypoosmotic Test, while the mitochondrial membrane potential was evaluated through the JC-1 fluorochrome. Data were analyzed by ANOVA/ Tukey test (P < 0.05). A significant decrease in progressive motility was observed in samples treated with or without Trolox during the refrigeration process, compared with fresh semen (P < 0.05). The progressive motility in fresh semen was 85.71 ± 1.74. No differences were observed in progresive motility between the samples refrigerated with and without Trolox. An increase in sperm vitality was observed on days 1 and 5 after refrigeration with Trolox compared with fresh semen (P < 0.05). The difference between the percentage of acrosomal and membrane integrity in treated and untreated samples was not significant (P > 0.05). Samples refrigerated with the addition of Trolox presented a significantly lower percentage of pre-capacitation on day 5 post-refrigeration compared with those which were not treated (P < 0.05). The mitochondrial membrane potential was 80.00 ± 4.69% prior to refrigeration and samples treated with Trolox (69.00 ± 7.75%, 58.75 ± 6.80% and 46.33 ± 12.66% at day 1, 2 and 5, respectively) presented higher values compared with the untreated ones (P < 0.05). The addition of Trolox to swine seminal samples produced an increase in their vitality as well as a decrease in the percentage of pre-capacitation and maintained the plasma membrane integrity while producing a decrease in sperm motility with a tendency to maintain a low intracellular redox state due to a low mitochondrial membrane potential.