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This study aimed to define sperm membrane protein markers of semen freezability of boars with the aid of a proteomic approach. Semen from fourteen adult boars were subjected to slow freezing and rapid thawing. After thawing, sperm vigor and motility were analyzed, and based on these results, animals were separated into two groups: good (GFEs) and poor freezability (PFEs). Sperm membrane proteins were extracted and subjected to two-dimensional electrophoresis. Stained gels were analyzed by computerized resources to indicate differentially expressed protein spots, that were identified by mass spectrometry.
Six animals showed good freezability with average sperm vigor and motility of 2.2 ± 0.8 and 41.8 ± 22.9, respectively, whereas eight boars showed poor freezability, with 1.9 ± 0.6 and 26.8 ± 17.5 of sperm vigor sperm motility, respectively. An average of 263 ± 62.2 spots per gel and 234.2 ± 54.6 of spots consistently present in all gels were detected. The intensities of five spots were significantly different between groups. Fc fragment of IgG binding protein and lactadherin were more intense in the PFE group, while Arylsulfatase A and F-actin capping protein subunit alpha 1 were more expressed in the GEF group. Fc fragment of IgG was higher in sperm membrane of poor freezability boars agglutinating sperm and decreasing mobility. Lactadherin was higher in sperm membrane of poor freezability boars possibly, leading to gamete oxidative stress. Arylsulfatase A was higher in sperm membrane of good freezability boars, playing role in post-thawing energy metabolism. F-actin capping alpha 1 was higher in sperm of good freezability boars, binding to actin filaments of the sperm tail.
Based on their functions and interactions with other proteins, we conclude that these four sperm membrane proteins may act as potential markers of boar semen freezability.
Daianny B.Guimarães, Tatyane B.Barros, Maurício F.van Tilburg, Jorge A.M.Martins, Arlindo A.Moura, Frederico B.Moreno, Ana C.Monteiro-Moreira, Renato A.Moreira, RicardoToniolli. Sperm membrane proteins associated with the boar semen cryopreservation. Animal Reproduction Science. Volume 183, August 2017, Pages 27-38. https://doi.org/10.1016/j.anireprosci.2017.06.005
