Effetc Of LDL Of Pigeon Eggs On HSP70Protein And Apoptosis Related Gene Expression In Cold Shock And Freeze-Thawed Boar Spermatozoa

Boar sperm is very susceptible to cold shock damage, and sensitive to peroxidative damage, because of sperm membrane phospholipids containing a large amount of unsaturated fatty acids and Semen itself is relatively low antioxidant capacity. The main purpose of this study was Evaluation of Boar semen during cryopreservation, supplementation different concentrations of pigeon eggs LDL in Boar semen, When frozen-thawed that SDS-PAGE and Western Blot and PCR_amplification and DNA electrophoresis were used to research the influence of it to The characteristics of Boar sperm vitality:curvilinear velocity (VCL)、 straight linear velocity (VSL)、average path velocity (VAP)、sperm acrosome intact, expression of heat stress protein (HSP70) gene and expression of anti-apoptotic(Bcl-21and Bcl-xl)genes and pro-apoptosis Bak). Results as follow:1、Whether cold shock or frozen-thawed, The characteristics of Boar sperm vitality:curvilinear velocity (VCL)、straight linear velocity (VSL)、average path velocity (VAP) the group of supplementation pigeon eggs LDL were exceed the group of did not supplementation pigeon eggs LDL; The effect of the cold shock group of Supplement9%LDL、frozen-thawed sperm group of Supplement9%LDL(-196℃) was best.2、Coomassie brilliant blue was used for assessment of cold shock and frozen-thawed Sperm acrosome integrity, result showed that sperm acrosome integrity of the cold shock group of supplementation pigeon eggs9%LDL and the frozen-thawed group of9%LDL(-196℃) were best.3、10%SDS-PAGE and Western Blot were used to analyze Different processed groups samples. result showed that With the decreasing temperature, the expression of heat stress protein (HSP70) gene were rise, Supplement9%LDL(-196℃) frozen-thawed sperm is most similar to fresh sperm.9%LDL(-196℃)、9%LDL(-80℃)、3%LDL (-196℃) group were no significant difference. 4、PCR amplification was used to analyze Different processed groups samples of proapoptotic Bcl-xl genes, result showed that the amount of gene expression of fresh semen group were exceed Other treatment groups; the amount of gene expression of3%LDL(-196℃) supplemented group were exceed Other three group except of fresh semen group; but The amount of gene expression of3%LDL(-80℃)and9%LDL(-80℃),9%LDL(-196℃) group were No significant difference.5、PCR amplification was used to analyze Different treatment groups samples of anti-apoptotic Bcl-21genes, result showed that the amount of gene expression of fresh semen group were exceed Other processed group; the amount of gene expression of3%LDL(-196℃) supplemented group were exceed Other three group except of fresh semen group; The amount of gene expression of3%LDL(-80℃) group was lowest; The amount of gene expression of3%LDL(-80℃),3%LDL(-196℃)、9%LDL(-80℃)、9%LDL(-196℃) group were no significant difference.6、PCR amplification was used to analyze Different processed groups samples of proapoptotic Bak genes, result showed that the amount of gene expression of fresh semen group were below Other treatment groups; the amount of gene expression of3%LDL (-80℃) supplemented group were below Other three group except of fresh semen group; The amount of gene expression of3%LDL (-196℃) group was maximum; The amount of gene expression of fresh semen group、3%LDL (-80℃)、9%LDL (-80℃)、9%LDL (-196℃) group were No significant difference.In conclusion, supplemented pigeon eggs LDL in boar semen extender during cryopreservation had a positive effect on post-thawed sperm survivability.