精子密度、R848浓度和孵育时间对奶山羊精子分选效果的互作影响OA

Immunological separation is a new method to achieve sex control.X and Y sperm can be sep-arated by direct activation of differential proteins,but the sorting system is still imperfect and the separa-tion effect is unstable.In this study,fresh semen from 10 dairy goats was collected.The effects of sperm density(3 × 108,6×108,9 × 108 cells/mL),concentration of activator R848(0,0.2,0.6,1.0,1.4,1.8μmol/L)and incubation time(30,45,60 min)on the isolation of X and Y sperm of dairy goats were ana-lyzed.The proportion of sperm after sorting was detected by flow cytometry.The results showed that,compared to the conventional diluted semen group(X-sperm proportion:51.4％),the proportion of X-sperm was significantly increased(P＜0.01)when collecting the lower stream of semen.Specifically,un-der sperm densities of 3× 108,6 × 108,and 9×108 cells/mL,with the addition of 1.4 μmol/L R848 and twice incubation for 60 min,30 min,and 45 min respectively,the X-sperm proportions reached 87.9％,88.1％,and 84.4％,representing increases of 36.5％,36.7％,and 33.0％compared to the control group.Similarly,compared to the conventional diluted semen group(Y-sperm proportion:51.0％),the propor-tion of Y-sperm was also significantly increased(P＜0.01)when collecting the upper stream of semen.At a sperm density of 6×108 cells/mL with either 1.4 μmol/L R848 incubated for 30 min or 0.6 μmol/L R848 incubated for 45 min,as well as at a sperm density of 9×108 cells/mL with 0.6 μmol/L R848 incubated for 60 min,the Y-sperm proportions were 83.7％,85.7％,and 84.4％,respectively.These corresponded to increases of 32.7％,34.7％,and 33.4％compared to the control group.Under suitable interaction condi-tions,the sperm TLR7/8 sorting system of dairy goats can obtain high-purity sex-controlled semen,and its accuracy reaches 85％,therefore,the goat sperm TLR7/8 sorting system is preliminarily established to improve the sex control technology of dairy goats based on immunological separation.