Effect of trolox added to freezing extenders over goat and ram spermatozoa

Lúcia Cristina Pereira Arruda,Ellen Cordeiro Bento Da Silva,Girliane Regina Da Silva,André Mariano Batista,Giovanna Isabella De Souza Couto,Helder Melo De Souza,Maria Madalena Pessoa Guerra,Tânia Maria Sarmento Da Silva

doi:10.33448/rsd-v10i5.14764

Lúcia Cristina Pereira Arruda, Ellen Cordeiro Bento Da Silva + Show 6 more

Open Access

https://doi.org/10.33448/rsd-v10i5.14764

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Abstract

Semen cryopreservation is responsible for decrease the gamete fertility, due to structural and functional damages. Among the various causes, oxidative stress, resulting from the higher generation of reactive oxygen species (ROS), has been attributed to affect semen quality. Thus, it was objectified evaluate the effect of Trolox on ram and goat sperm, subjected to freezing. Semen pools of goat (n=5) and ram (n=6) were diluted in skimmed milk (7% glycerol) or Tris-egg yolk (5% glycerol) extender, respectively, added or not of Trolox (0, 20 or 40 μM/ml) and frozen. After thawing (37 °C/30 s), aliquots of semen were evaluated for lipid peroxidation by high performance liquid chromatography, coupled with a photodiode array detector (HPLC-DAD), and flow cytometry (C11-BODIPY581/591), besides of plasma membrane and acrosome integrity by fluorescence microscopy, and sperm kinetics by computerized sperm analysis (CASA). The antioxidant treatment with Trolox did not determine significant effects (p>0.05) on lipid peroxidation, plasma membrane integrity, acrosomal integrity and on the kinetic parameters evaluated. Thus, it is concluded that Trolox (20 or 40 μM) did not have a protective or deleterious effect on goats and ram sperm, submitted to freezing.

Highlights

The semen cryopreservation process determines cellular changes that contribute to the fertility reduction, when compared to fresh semen (Bicudo et al, 2007), being the plasma membrane one of the most affected structures (Castro et al, 2016)
Based on the exposed above, was objected in this study evaluated the Trolox effect on goat and ram cryopreserved sperm, through the lipid peroxidation, membranes integrity and kinematics evaluation
The results demonstrated that the Trolox addition, at 20 or 40 μM concentrations, to the goat and ram semen cryopreservation extender does not determine significant differences (p>0.05) in the levels of MDA or in the percentage of cells marked with C11-BODIPY581/591, when compared to the control group (Table 1)

Summary

Introduction

The semen cryopreservation process determines cellular changes that contribute to the fertility reduction, when compared to fresh semen (Bicudo et al, 2007), being the plasma membrane one of the most affected structures (Castro et al, 2016). These changes in sperm cells are associated with the biochemical, osmotic, thermal and mechanical stresses, which are seen at different stages of freezing process (Gangawar et al, 2016). The evaluation and control of the oxidative status and antioxidant defenses system is important as an indicator of the male fertility, especially (Colagar et al, 2013)

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