Identification of sperm-specific proteins associated with high fertility of Punjab Red and RIR x Local cross roosters with SDS-PAGE and Immunoblotting

The present study aimed at characterization of fertility-associated proteins in PR and RIR x Local 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. SDS-PAGE of sperm extracts of PR and RIR x Local cross breeds resulted in resolution of 12 and 23 proteins on 12% acrylamide gels and anti-SP IgG reacted only with 8 and 9 sperm proteins of PR and RIR x Local cross roosters on immunoblots. The SDS-PAGE and immunoblotting analysis also indicated a variation in sperm proteins among two breeds and high/low fertile roosters. It can be concluded that the selection of roosters on the based on proteins of 65/ 25; 70/ 46/ 30 kDa may be specifically associated with high fertility of PR and RIR x Local cross, respectively. The proteins 62 kDa (PR) and 40kDa (RIR x Local cross) may be specifically responsible for low fertility.