Proteomic comparison of non-sexed and sexed (X-bearing) cryopreserved bull semen

It is widely recognized that quality of spermatozoa in sexed semen (SS) samples is not as great as for conventional, non-sexed semen (NS). There are differences in qualitative and biochemical variables between spermatozoa in NS and SS. Information, however, is lacking on molecular differences, especially concerning spermatozoa proteomic differences is SS and NS. The objective of this study was to compare commercially available NS and SS bull semen by evaluating sperm quality variables in conjunction with a proteomics approach. Results from flow cytometry and computer-assisted sperm analyses indicated there was less sperm motility, viability, mitochondrial potential and acrosome integrity in sperm from SS. Results from proteomic analysis indicated sperm from NS and SS samples were characterized by different protein profiles. There was identification of 70 sperm proteins that differed in abundance and six protein spots that were different in extent of carbonylation. Sperm from SS had altered structures of enzymes involved in glycolysis, oxidative phosphorylation and maintenance of a constant adenylate energy charge. Furthermore, sperm from SS had alterations of several flagella substructures, especially outer dense fiber proteins, which were less abundant and more carbonylated than in sperm from NS. In sperm of SS, compared with NS, there were differences in abundance of proteins involved in capacitation, acrosome reaction and sperm-egg fusion as well as lesser abundances of sperm surface proteins. Results enable a greater understanding of differences between sperm from NS and SS samples, thereby contributing to development of improved protocols for more effective protection of sexed spermatozoa during processing.