Investigation Of Sperm Fructose Metabolism During Liquid Storage Pig Semen

Use liquid diluent to extend the pig semen save time for the promotion of the pig in vitro fertilization and good pedigree breeding pig is critical, but the use of diluent preservation process will encounter some problems: add ingredient and the adding amount of diversity preservation solution; Sperm survival time is shorter in the preservation solution; Fertilization rate low. Solve the seminal fluid in the process of the liquid to save save the effect not beautiful can be from two aspects, one is the sperm in the process of metabolic energy supply, second it is timely remove sperm generation of harmful substances in the metabolic process. Furthermore, the existence of the PPP pathway and need for the Krebs cycle in sperm of different species has yet to be confirmed. Therefore, studies on sperm fructose metabolism during liquid storage of pig semen are important not only for enriching sperm energy metabolism but also for improving the technique of liquid semen storage.The objective of this project is to study sperm glucose utilization and metabolic pathways during liquid semen storage. To this end, boar semen will be preserved at 15℃ in the basic PBS extender that does not contain any energy substrates but supplemented with fructose, intermediate metabolism of glucose and /or inhibition of glucolysis, PPP or mitochondria electron transport. Data of sperm motility, viability and acrosome integrite, changes of antioxidant capacity and mitochondrial membrane potential at different time of preservation obtained will be also used to formulate an optimal system that provided the best energy for extended liquid storage of boar semen. The result of this study indicate thatMotility and viability of boar sperm is were highest if he semen is stored in PBS supplemented with 3mM fructose when the semen is storaged at 15℃, and a sperm motility of 30% motility for 5 days.Motility and viability of boar sperm is were highest if he semen is stored in PBS supplemented with 5mM pyruvic acid when the semen is storaged at 15℃, and a sperm motility of 30% motility for 5 days.Compared with control group that in adding 3mM fructose conditions, the save time, motility and viability and mitochondrial function integrity of boar sperm was significant reduced in the dilution of sodium iodoacetate which is inhibitor of glycolytic pathway.Compared with control group that in adding 3mM fructose conditions, the save time and motility of boar sperm was not effected in the dilution of 6-amino-nicotinamide, inhibitor of pentose phosphate pathway. But antioxidant capacity significant decline.Compared with control group that in adding 3mM fructose conditions, the save time of more than 30% motility and motility of boar sperm was remarkable declined in the dilution of rotenone, inhibitor of electron transport chain in oxidative phosphorylation.Through this study, the following conclusions: 1) Under 15 ℃, PBS save diluent to add 3 mm fructose can be beneficial to the pig sperm to survive, to join the large amounts of fructose will reduce the pig sperm in vitro preservation time;2) pig sperm not only can use fructose metabolism, also can use the intermediate with pyruvic acid in the process of sugar metabolism; 3) the glycolytic pathway is to maintain the pig sperm vitality, maintain the important guarantee of mitochondrial function is not affected;4) The motility of boar sperm is not effected by pentose phosphate pathway but antioxidant capacity can be enhanced; 5) Aerobic oxidation is beneficial to the maintenance of the pig sperm motility.