Identification of new protein biomarkers associated with the boar fertility using iTRAQ-based quantitative proteomic analysis

In this study, we performed the isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis in the spermatozoa of Landrace boars with different fertility potentials and investigated the ability of sperm acrosome associated 4 (SPACA4) and IZUMO family member 2 (IZUMO2) to predict the reproductive perform of boars. The iTRAQ results revealed that 202 proteins were up-regulated and 43 proteins were down-regulated in the spermatozoa from high fertility boars. SPACA4 and IZUMO2 protein levels were significantly up-regulated in the spermatozoa from high fertility boars. SPACA4 and IZUMO2 expression were specifically detected in the adult boar testis. SPACA4 levels were positively correlated with Sow's farrowing rate and reproductive efficiency, but not litter size. IZUMO2 were positively correlated with litter size, Sow's farrowing rate and reproductive efficiency. Treating the boar semen with SPACA4 or IZUMO2 antibodies for 30 min and 60 min failed to affect the sperm motility; while treating the semen with SPACA4 antibody significantly reduced the fertilization and cleavage rates. Similar results for fertilization and cleavage rates were found in IZUMO2 antibody-treated semen. Collectively, our results indicated that protein levels of SPACA4 and IZUMO2 in the spermatozoa were positively related to the reproductive performance of Landrace boars.