Effects Of Low Temperatures,Capacitation And Low-Density Lipoprotein Of Pigeon Eggs On Cytoskeleton Integrity In Boar Spermatozoa

The main purpose of the research was to detect the effects of capacitation and low temperature on Integrity of the cytoskeleton in boar sperm, including tubulin and actin which were the main compositions of cytoskeleton and tried to explain the reason why motility decreased dramatically after low temperature cytopreservation in biology molecular field. LDL extracted from pigeon eggs was used as reagent instead of yolk to detect its effect on cytoskeleton. SDS-PAGE and Western Blot were used to detect the content changes of tubulin and actin after capacitation and low temperature. Results as follow:1. CM was used to capacitate and incubate4hours at38℃,5%C02. SDS-PAGE was used to analyse the content of tubulin and actin. Result showed that content of tubulin did not change between capacitation and fresh boar sperm, but actin decreased slightly.2. SDS-PAGE was used to analyze changes of kinds and content of protein, result showed that kinds and content of some protein changed after low temperature treatments, but no difference after capacitation.3. Boar sperm treated by low temperature24hours and Western Blot was used to analyze the content changes of tubulin and actin. Rsesult showed that tubulin decreased slightly but actin decreased dramatically.-196℃was better than-80℃. 4. LDL extracted from pigeon eggs was used as reagent instead of yolk to protect boar sperm. The result of Western Blot showed that LDL groups were better than yolk groups in both low temperatures, and no significant difference between temperatures.5. Sperm motility and acrosome integrity and membrane integrity rate were detected, result showed that LDL groups were better than yolk groups in all above.