Characterization of fertility associated sperm proteins in Aseel and Rhode Island Red chicken breeds

Abstract

This study focused on characterization of fertility associated proteins in Aseel and RIR roosters and was conducted on two generations of birds. Roosters were divided into high (>50%) and low fertility groups (<50%) based on sperm function tests and fertility rate in both the generations. Polyclonal antibodies were raised in rabbits against sperm proteins of first generation highly fertile roosters and tested for characterization of fertility associated sperm proteins in the second generation of same roosters. IgG-fraction against proteins (Anti-SP-IgG) was reacted with sperm proteins of both high and low fertile roosters of second generation on immunoblots. Sperm proteins present in highly fertile roosters were further characterized by Mass Spectrometry (MS). Use of SDS-PAGE for evaluation of sperm extracts of Aseel and RIR breeds resulted in resolution of 16 and 10 proteins on 12% acrylamide gels. Anti-SP-IgG reacted with eight and ten sperm proteins of Aseel and RIR roosters on immunoblots. The SDS-PAGE and immunoblotting analysis also indicated a variation in sperm proteins among two breeds and high/low fertile roosters. The MS analysis indicated matching of 20, 30, and 20, 25 kDa proteins (associated with high fertility rate) of Aseel and RIR roosters with immunoglobulin kappa chain variable, phospholipase A2 (PLA2), hypothetical N332-08551 partial and cystatin like partial proteins with a top score of 41, 46, 52 and 43, respectively. Considering the function and importance of matching proteins in male reproduction, these proteins may be further explored as potential markers for fertility evaluation of Aseel and RIR roosters.