Effect of Glutathione Supplementation in Cryoprotectant Modification on Tyrosine Phosphorylation, Acrosin Expression and Acrosome Reaction of Post-Thawing Spermatozoa Quality

Abstract

Background: Tyrosine phosphorylation, acrosin, and acrosome reaction play an important role in fertilisation. However, cryopreservation causes changes in tyrosine phosphorylation, acrosin expression, and acrosome reaction which affect the quality of spermatozoa. Cryoprotectant media added with antioxidants is needed to protect Spermatozoa from the effects of cryopreservation so that the quality of spermatozoa can be maintained. Objectives: This research examined the effect of glutathione (GSH) supplementation in cryopreservation media on tyrosine phosphorylation, acrosin expression, and acrosome reaction. In this research, pure modified Cryoprotectant (CPA) was compared with CPA supplemented with GSH in three different concentrations. Materials and Methods: The research sample was male mus musculus albinus strain Deutchland Denken Yoken (DDY). Mice spermatozoa was cryopreserved and several parameters were measured including tyrosine phosphorylation, acrosin expression, and acrosome reaction. Results: The addition of GSH to the modified CPA increased tyrosine phosphorylation, acrosin expression, and acrosome reaction (maintaining acrosome integrity). The group with 1.00 mM GSH obtained the highest results among the other groups. Significant increases were found in tyrosine phosphorylation, acrosin expression, and acrosome reaction after the addition of 1.00 mM GSH. Conclusion: Glutathione supplementation in modified CPA can increase tyrosine phosphorylation, acrosin expression, and acrosome reaction of frozen-thawed spermatozoa. Treatment using GSH at a dose of 1.00 mM is the most effective and modification of CPA with the addition of glutathione can improve the tyrosine phosphorylation, acrosin expression and acrosome reaction in cryopreserved spermatozoa. Keywords: spermatozoa cryopreservation, CPA modification, glutathione (GSH), tyrosine phosphorylation, acrosin expression, acrosome reaction