Diluent pH affects sperm motility via GSK3 α/β-hexokinase pathway for the efficient enrichment of X-sperm to increase the female kids rate of dairy goats

Abstract

Dairy goats are the goats bred with the ability to produce large quantities of milk, and the increase of the female kid rate of breeding dairy goats is beneficial for milk production and economic benefits of dairy goat farms. Our previous study revealed that regulating the pH of dairy goat semen diluent to 6.2 or 7.4 respectively, the proportion of X chromosome bearing sperm (X-sperm) in the up and down layers of the tube after incubation was significantly higher than that of Y chromosome bearing sperm (Y-sperm) i.e. enriched X-sperm. In this study, fresh dairy goat semen collected in different seasons was diluted in different pH solutions to calculate the number and rate of X-sperm and to measure the functional parameters of enriched sperm. The artificial insemination experiments were performed with enriched X-sperm. The mechanisms of regulating the pH of diluent affecting sperm enrichment were further studied. The results showed that the proportion of enriched X-sperm in pH 6.2 and 7.4 diluents of sperm collected in different seasons showed no significantly different, but were significantly higher than that of the control group (pH 6.8). The in vitro functional parameters of X-sperm enriched in pH 6.2 and 7.4 diluent solution were not significantly different from those of the control group (P > 0.05). After artificial insemination with X-sperm enriched in pH7.4 diluent, the proportion of female offspring was significantly higher than that of the control group. It was found that the regulating pH of the diluent affected sperm mitochondrial activity and glucose uptake capacity via phosphorylating NF-κB and GSK3α/β proteins. The motility activity of X-sperm was enhanced under acidic conditions and weakened under alkaline conditions, which was conducive to the effective enrichment of X-sperm. This study demonstrated that the number and proportion of X-sperm enriched using pH 7.4 diluent were elevated, and the proportion of female kids was increased. This technology can be used for the reproduction and production of dairy goats in farms at large scales.