Proteomic identification of boar seminal plasma proteins related to sperm resistance to cooling at 17 °C.

The modern pig industry relies on extensive use of artificial insemination with cooled semen. It is important that semen doses maintain their quality during processing, transport and storage before insemination to guarantee maximum fertility rates. However, ejaculates may respond differently to liquid preservation at 17 °C, despite the optimal quality assessed before cooling. Thus, the aim of this study was to identify differences in seminal plasma proteome of ejaculates with a higher or lower seminal resistance to storage at 17 °C. A total of 148 ejaculates from 65 sexually mature healthy boars were classified as: High Resistance to cooling (HR, total motility > 60% at 144h) and Low resistance to cooling (LR, total motility <60 at 72h). To identify differentially expressed seminal plasma proteins between HR and LR ejaculates, ten ejaculates of each group were analyzed by 2D SDS-PAGE and ESI-Q-TOF mass spectrometry. The proteins associated with HR ejaculates were cathepsin B (spot 2803 and 6601, p < 0.01); spermadhesin PSP-I (spots 3101 and 3103, p < 0.05); epididymal secretory protein E1 precursor (spot 2101, p < 0.05) and IgGFc binding protein (spot 1603, p < 0.01). The protein associated with LR group was the Major seminal plasma PSPI (spot 9103, p < 0.01). To our knowledge, this is the first report of the association of boar seminal plasma proteins to semen resistance to cold storage at 17 °C. These results suggest the use of these proteins as biomarkers for semen resistance to preservation at 17 °C.

Seminal plasma proteins and their relationship with sperm motility and morphology in boars.

The identification of biomarkers associated with seminal traits could aid in the selection of higher quality ejaculates and benefit the swine industry. The objective of this study was to identify boar seminal plasma proteins associated with sperm motility and morphology. Twenty ejaculates from fifteen adult boars from a commercial boar stud were used for this work. After routine semen collection and analysis, ejaculates were classified into two groups: high-quality semen (HQS) and low-quality semen (LQS), based on sperm motility and morphology. Semen samples were processed for seminal plasma separation and analysis by 2D SDS-PAGE. Total and progressive sperm motility differed between groups (p < 0.001), as well sperm morphology (p < 0.05). The intensity of spots identified as Major seminal plasma PSP-I (PSP-I) and cathepsin B (CTSB) was higher in LQS as compared to HQS samples (p < 0.05). Also, PSP-I was positively associated with major and sperm cauda defects. Sperm motility was negatively correlated with both PSP-I and cathepsin B. We conclude that high concentrations of Major seminal plasma PSP-I and cathepsin B in boar seminal plasma are associated with reduced total and progressive sperm motility and low sperm morphology and might be used as biomarkers for semen quality.