Studies On Proacrosin Kinetics Activation In Boar Spermatozoa

The main purpose of this experiment was to research freeze-thaw and differences after pre-acrosome were actived into acrosome. Hold sperm at 4℃respectively in 20min,4min,60min,80min,100min, choose the best time for porcine sperm to freeze; and added glycerol in concentration of 2%,4%,6%,8% to the best one, Screening the most suitable concentration of glycerol for pig sperm; selected the most suitable cryoprotectants for pig sperm,and those take a step forward to optimization of pig sperm freezing and thawing process. Analyze the transformation process of pre-acrosome in freeze-thaw and capacitation by SDS-PAGE and Western Blotting. The results are as follows:1,When sperm got balanced at 4℃, the porcine sperm quality which cool balanced in 60min were significantly higher than those in 20min,40min,80min, 100min (P<0.05), sperm quality generated peaks at this time, sperm motility rate was 33.75%.Acrosome integrity rate was 34.25%; hypotonic swelling rate was 31.50%.moreover, sperm quality in treatmental group were significantly higher than control group, thus we can see the importance of balance in condition of 4℃in the process of sperm frozen.Compared 2% and 4% glycerol to sperm pretreatment group, quality of pig sperm were significantly higher than control group and other concentration groups after freezing and thawing (P<0.05), the live rates were 36.2% and 35.5%:acrosomal integrity rates were 35.4% and 35.2%; hypotonic swelling rates were 33.8% and 34.2%. It Showed that a concentration of 2% and 4% of glycerol in the freezing and thawing process can more effectively protect the sperm, improving the quality of sperm. enhancing the value of actual use of frozen-thawed sperm.2,With 5mg/mL Caffeine and 200mg/ml vitamin E as cryoprotectant for freezing and fresh sperm respectively to proceed SDS-PAGE analysis and comparison,Adding caffeine and vitamin E as cryoprotectant for freezing and thawing sperm showed less bands compared to frozen-thawed sperm samples of caffeine-free,Therefore. more similar with the fresh sperm.Such results indicate that caffeine and vitamin E can prevent the transformation of individual proteins in advance. However, caffeine is more effective after comparing.3,In frozen-thawed process, some former acrosomal proteins of boar sperm were stimulated and destroyed, a proportion of 43KD former acrosomal proteins were activated and transformed to 35KDa and 28KDα-andβacrosomal proteins.