Effect of Fractionated Seminal Plasma on Sperm Characteristics following Cryopreservation of Boar Semen

Abstract

This study aimed to investigate the effect of fractionated seminal plasma (SP) on boar sperm characteristics following cryopreservation. Gel filtration chromatography yielded two fractions: SP1 with more than 40 kDa (>40 kDa) and SP2 with less than 40 kDa (<40 kDa). The fractionated SP (SP1 and SP2), whole seminal plasma (wSP) and Beltsville Thawing Solution (BTS) were used for the treatment of semen before freezing-thawing. Besides the analysis of sperm motility characteristics, plasma membrane integrity (PMI), acrosome integrity, and viability (Vybrant Apoptosis Assay) were analyzed in pre-freeze and post-thaw (PT) semen. Among the analyzed pre-freeze sperm parameters, rapid movement was markedly affected by boar and treatment. Furthermore, boar and treatment were significant sources of variations in PT semen quality. Treatment with wSP caused a marked reduction in PT semen quality compared with BTS, SP1 or SP2. Wide variations in PT acrosome integrity and viability were observed in spermatozoa treated with BTS and the fractionated SP, being significantly higher in the SP1-and SP2-treated samples. However, PT semen quality did not differ between semen samples treated with SP1 and SP2. Representative electrophoretic profiles of sperm proteins from each treatment showed quantitative and qualitative differences, indicating varying effects of the cryopreservation procedure on the sperm membrane integrity. The findings of this study indicated that the cryoprotective effects of the SP components varied among boars and that different components of the fractionated SP exerted varying effects on sperm functions following cryopreservation. It could be suggested that the variable protective protein components of either fractionated SP ameliorated alterations in the sperm membranes during cryopreservation, resulting in reduced susceptibility to cryo-damage.