Studys On The Modification Of Ubiquitination And Sumoylation Of Ai In Capacitated And Cryo-capacitated In Boar Sperm

Capacitation gives sperm the ability to enhance activity,it enables the sperm to interact with the oocyte zona pellucida(ZP),then the fertilization process is completed.There are many similarities between capacitation and cryo-capacitation.Studies have shown that protein phosphorylation during sperm capacitation is a very important event.Capacitation may be regulated by the Ubiquitin-proteasome system(UPS),and studies also have shown that small ubiquitin-related modifier(SUMO)is associated with phosphorylation and Ubiquitination.Therefore,the purpose of this study was to explore the differences in the modification levels of ubiquitination and sumoylation of acrosomal enzyme inhibitor(AI)in capacitated and hypothermic capacitated boar sperm,to further understand the protein modification and degradation in the prcess of sperm capacitation,to provide the basis for improving the quality of pig sperm after frozen-thawed.Duroc boar semen was selected in this study,the subjects were divided into four groups:fresh semen(control group),capacitation treatment,inducing acrosome reaction treatment and cryo-capacitation treatment.Detection was performed by CASA,SDS-PAGE and Western blot,each group was repeated five times.The results showed that:1.Sperm protein tyrosine phosphorylation levels of the capacitation treatment,inducing acrosome reaction treatment,cryo-capacitation treatment groups were significantly higher than those in the control group(P<0.05);2.The expression of sperm acrosin inhibitor of the capacitation treatment,inducing acrosome reaction treatment,cryo-capacitation treatment groups were significantly lower than that in the control group(P<0.05);3.The ubiquitin expression level of sperm in capacitation and inducing acrosome reaction groups was significantly higher than that in control group(P<0.05),no tyrosine phosphorylation of sperm protein was observed after adding E2 inhibitor;4.The sumoylation expression level of sperm in capacitation and inducing acrosome reaction groups was significantly higher than that in control group(P<0.05),and increased,with the increase of 2-D08 inhibitor concentration,the tyrosine phosphorylation level of sperm protein in each group decreased gradually and was lower than that of control group.The conclusions of this study are that:1.Boar sperm capacitation was accompanied by the increase of sperm protein tyrosine phosphorylation,the enhancement of ubiquitination and ubiquitination modification,and the degradation of acrosomal enzyme inhibitors;2.In vitro inhibition of ubiquitination or sumoylation hindering boar speim capacitation;3.Ubiquitination and sumoylation modification may be used as markers of boar sperm capacitation.