Study on the Separation Effect of Pig X,Y Sperm of Different Acid-Base Diluents Combined with Creatine Monohydrate
The study aimed to explore the separation effect of porcine X,Y sperm by adjusting the acidity and alkalinity of porcine semen diluent combined with creatine monohydrate(CMH).In this study,eight healthy Duroc boars aged 2-4 years were used to collect semen.By adjusting the pH of semen diluent Beltsville(BTS),the optimal pH of the X,Y sperm sorting system was de-termined by sperm computer-aided analysis system(CASA),and the optimal concentration of creatine monohydrate was determined by CASA.The effect of X,Y sperm sorting was evaluated by immunofluorescence staining and flow cytometry,and sperm quality indexes such as acro-some,plasma membrane integrity,and reactive oxygen species content(ROS)were detected by fluorescence labeling and MitoSOX Red staining of sperm ROS.The results showed that when the pH of the diluent was 6.4 and 7.1,there was no significant effect on sperm viability after in-cubation for 60 min(P＞0.05),and the proportion of sperm in the upper layer moving forward was significantly higher than that in the lower layer(P＜0.01).Adding 750 µmol·L-1 creatine monohydrate to the diluent and incubating for 60min significantly improved sperm motility,linear movement rate,curve movement rate,and wobble(P＜0.05).Using acid diluent com-bined with creatine monohydrate to separate pig X,Y sperm,the proportion of X sperm in the upper semen after separation was 63.32％.Alkaline diluent combined with creatine monohydrate was used to separate pig X,Y sperm.After separation,the proportion of Y sperm in the upper semen reached 67.19％.Further study showed that there was no significant difference in acro-some integrity,plasma membrane integrity,and ROS level between the two groups(P＞0.05),while ATP level,mitochondrial membrane potential,and exercise performance were significantly higher than those of the control group(P＜0.05)after separation.To sum up,this study estab-lished a simple and effective separation system of X and Y sperm in pigs by adjusting the acidity and alkalinity of the diluent in parallel with creatine monohydrate.This method did not affect the quality of sperm and improved the motility of sperm after separation,which further provided a theoretical basis for the research and development of new technologies for controlling semen in pigs.
pig spermacid and alkaline diluentcreatine monohydratesexually controlled se-menX,Y sperm sorting
万方数据
维普
