Cryopreservation And Cryopreservation Of Donkey Semen

The research on the preservation of various mammalian semen has reached a high level and has been widely promoted.The semen semen preservation technique is still not mature enough in the experimental stage.The actual production and reproduction still adopts the method of artificial insemination after dilution or dilution of fresh essence.This study attempted to use semen graphene oxide(GO)as a semen protective substance for semen semen preservation in order to explore the method of semen semen preservation.GO is a product of graphene after oxidation and is amphiphilic.According to the carbon skeleton structure unique to GO,it was found that GO not only effectively inhibited recrystallization and ice crystal growth,but also modified the morphology of ice crystals,forming a curved surface between GOs and inhibiting ice crystal growth by Gibbs-Thompson effect.In this study,basal solution(Tris30.4 g/L,citric acid 17.0 g/L,glucose 15.0 g/L,fructose 15.0 g/L,penicillin 0.6 g/L,streptomycin 1.0 g/L)+15% yolk As a diluent,the relative optimum amount of graphene oxide added was determined by cryopreservation,followed by comparison of different pretreatment effects,comparison of different freezing procedures,comparison of fumigation height and fumigation time of liquid nitrogen steam fumigation,and comparison of different thawing methods.I hope to get an economical,simple and effective freezing method.The results showed that the sperm motility,plasma membrane integrity rate and acrosome integrity rate of the basal solution +15% egg yolk + 10% graphene oxide were significantly higher than other groups(p<0.01),and the deformity rate was significantly lower than other.Group(p<0.01).Before cryopreservation,the two pretreatment methods: direct centrifugation and dilution five times centrifuged for freezing,there was no significant difference in sperm motility,deformity rate,plasma membrane integrity rate and acrosome integrity rate after thawing(p>0.05),but Comparing the sperm quality index,it was found that the direct centrifugation method(0.52)was better than the dilution five-fold centrifugation method(0.50);the sperm motility of the p1 group was significantly higher than that of the other two groups after the freeze-thaw method(p<0.01),and other indicators were not.Significant difference(p>0.05);when steaming by steam fumigation,the sperm motility was significantly higher than the other two groups(p<0.01)when the height of the liquid nitrogen surface was 4cm,and 10 min after the liquid nitrogen surface was fumigation for 10 min.The sperm motility rate was significantly higher than the other two groups(p<0.01);comparing the 37°C 30 s thawing and 60°C 6s thawing methods,the 60°C 6s thawing method sperm thawing activity was significantly higher than the 37°C 30 s thawing method(p<0.01),there was no significant difference between other indexes(p>0.05).The expression of RPL31,PLP1 and PAPSS2 genes in fresh and frozen-thawed semen was detected by q RT-PCR.Compared with RPL31 The PLP1 gene was significantly down-regulated(p<0.01),and the PAPSS2 gene was significantly upregulated.(p<0.01).Some possibilities are provided for explaining the mechanism of sperm cryopreservation during cryopreservation.