Influence of antioxidants on dna intergrity, mitochondrial function and fertilizing potentials of cryopreserved buffalo seprmatozoa

Abstract EN

The objective of the current study was to evaluate the effect of cryopreservation and fortification of freezing extender with some antioxidants on the DNA integrity and the fertilizing potentials of cryopreserved buffalo spermatozoa.

Buffalo semen was collected, evaluated and extended in Tries-based extender supplemented with different concentrations of reduced glutathione (GSH), catalase (CAT), alpha lipoid acid (ALA) and superoxide dismutase (SOD).

Semen was examined post-thawing to evaluate, free ability, mitochondrial function, DNA integrity and the natural and assisted fertilizing potentials.

The main findings emerging from the present • study were that cryopreservation decreased (P<0.t) l) significantly sperm free ability, mitochondrial activity, DNA integrity and fertilizing potentials of frozen-thawed buffalo spermatozoa.

Meanwhile, in vitro provision of freezing extender with GSH, catalase, ALA and SOD had a beneficial effect on the function of the cryopreserved buffalo spermatozoa, in a dose dependent trend.

Fortification of semen extender with 10 mm GSH 100 U/ml catalase, 15 mm ALA or 50 U/ml SOD increased (P

Moreover, improved the mitochondrial function (3.42 ± 0.05, 3.29±0.16, 2.81 ± 0.09 and 3.31 ± 0.08, respectively); DNA integrity (37.67 ± 4.33, 36.33 ± 2.61, 43.33±5.05 and 36.00 ± 5.04%, respectively).

Additionally, in vitro provision of freezing extender with 10 mm GSH, 100 U / ml catalase or 50 U/ml SOD increased in vitro embryo development to the blastocyst stage (11.86, 11.54 and 14.20 %, respectively) and augmented the natural pregnancy rate (62.11, 62.79 and 67.74%, respectively).

In conclusion, cryopreservation promotes DNA fragmentation in buffalo spermatozoa that can be counteracted by the addition of GSH, catalase, ALA and SOD.

These antioxidants appear to play an important role in sperm antioxidant defense strategy in a dose dependent trend and could be of significant benefit in improving the free ability, DNA integrity and the natural and assisted fertilizing capacity of the cryopreserved buffalo spermatozoa.