Effect Of Trehalose On Cryopreservation Of Semen Of Cashmere Goat

Trehalose is a disaccharide with anti-oxidation function and non-permeable freezing protection function.It is widely used in the freezing of cells and semen,now.But its mechanism of action has not been fully elucidated,The purpose of this experiment is to study the effect of different concentrations of trehalose,different cooling rates,different combinations of trehalose and glycerol on the semen freezing of cashmere goats.At the same time,metabolomics was used to analyze the effect of trehalose on sperm cryopreservation from the metabolite level,and to study the mechanism of trehalose on Inner Mongolia cashmere goat sperm cryopreservation.The contents are as follows:1.The effects of different cooling rates on the cryopreservation of semen in Inner Mongolia Cashmere Goats.Compared with the traditional Tris-citric acid-glucose(TCG)extender,trehalose was added to the cashmere goat semen frozen extender,the optimal sperm cooling rate was 0.16?/min,and the sperm motility after freezing and thawing reached 58.3%.When the cooling rate of TCG is between 0.2-0.25?/min,the sperm motility is the highest,reaching 56.1%.Therefore,when trehalose is used to freeze cashmere goat semen,a slower rate should be used for cooling.When the cooling rate is maintained at 0.16?/min,Inner Mongolia cashmere goat semen is best frozen for preservation.2.The effects of different combinations of trehalose and glycerol on the motility,sperm structure and antioxidant performance of cashmere goat semen after freezing and thawing were compared.0 mM(T0),50 mM(T50)trehalose and 0%(G0),1%(G1),2%(G2),3%(G3)glycerol are combined into T0G0,T0G1,T0G2,T0G3,T50G0,T50G1,T50G2,T50G3.In the case of using TCG as the base solution,the combined use of trehalose and glycerin improves the effect of sperm cryopreservation through a synergistic effect,and the optimal concentration for its function is 50 mM trehalose and 1%glycerol.After sperm freezing and thawing,the sperm motility,acrosomal plasma membrane integrity rate and antioxidant performance were significantly higher than other groups(P<0.05).3.Metabolomics was used to analyze the effect of the addition of trehalose and the addition of trehalose-free extender on the cold storage of cashmere goat semen.The metabolites of the seminal plasma after freezing and thawing of the two groups were tested.The results showed that 309 differential metabolites were detected,with 242 differential metabolites up-regulated and 27metabolite down-regulated.The Variable important in projection(VIP),was used to further screen out 126 different metabolites,and there was a correlation between the metabolites.Among them,13 kinds of metabolites can be used as potential biomarkers,10 kinds of potential biomarkers are expressed in the trehalose group,and 3 kinds of potential biomarkers are expressed in the trehalose-free group.The metabolomic level can prove that trehalose reduces the damage of sperm during freezing by reducing ROS levels and inhibiting sperm capacitation in advance.