| The reproductive efficiency of Meishan,indigenous Chinese pig,is significantly higher than that of Duroc boar from the west.However,the underlying moulecular mechanism remains unclear.Actually,apart from the females,sperm quality and fertility ability also affect the conception rate and embryo survival rate.With the development of proteome,many proteins related to sperm motility,fertility ability and farrowing rate which can directly or indirectly influence conception rate or farrowing rate,were identified as reproductive biomarkers.However,few research has been reported if there exists a difference between Meishan and Duroc boar sperm,which thereby influence the reproductive rate.Therefore,the present study compared sperm quality,proteins and protein post-translational modifications between Meishan and Duroc boar spermatozoa to explore the relationship between fertility ability and specific proteins.Furthermore,the present work was hope to provide theoretical basis for the molecular mechanism underlying the difference of male fertility between indigenous Chinese pig and introduced pig to investigate.Three experiments were performed as follows.1.Meishan and Duroc boar sperm quality was compared during the perseveration under different temperatures(17 ℃ and 4 ℃)in vitro.Results showed that sperm motility and energy metabolism level in Meishan and Duroc boar sperm both declined,with the extension of preservation.Furthermore,the motility,ATP level,mitochondrial membrane potential,GAPDH activity and cAMP level in Meishan boar sperm were higher than that in Duroc while conserved at 17 ℃ and 4 ℃.Markedly,sperm motility of Meishan boar was higher than that of Duroc at the first day of preservation(P < 0.05).The results suggested that there may exist differences in cytoskeletal protein components and the expression of metabolism enzyme in sperm between Meishan and Duroc.2.In order to verify the predicted results,iTRAQ labeling technique coupled to LS-MS/MS analysis was used to identify and compare the sperm proteins in Meishan and Duroc.In the present study,1597 kind of proteins were identified and we found 190 kind of proteins differently expressed between Meishan and Duroc boar sperm.Bioinformation analysis showed that the expressed differential proteins including mitochondrial proteins,cytoskeletal proteins and metabolism enzymes,mainly involved in the regulation sperm motility,energy metabolism and fertilization.ODF2,LDHC,CALM,ACE,ENO1,positively related to the farrowing rate,were highly expressed in Meishan boar sperm.However,some proteins including NDUFS2,RAB2 A and some proteasomes,negatively related to the farrowing rate,were highly expressed in the Duroc boar sperm.3.To further validate the differently expressed proteins between Meishan and Duroc boar sperm,we used Western Blot to identify some specific proteins and compare the level of protein post-translational modifications(PTMS)of sperm in Meishan and Duroc.The results showing that SPAG6,ACE2 and CALM,positively related to reproductive ability,were highly expressed in Meishan boar sperm.However,the expression of NDUFS2,negatively related to reproductive ability,was lower in Meishan boar than that in Duroc sperm.Moreover,the levels of sperm protein serine/threonine phosphorylation,tyrosine phosphorylation,acetylation and succinylation in Meishan were higher than that in Duroc.Given to the results,the present study suggested that the highly expression of proteins such as mitochondrial proteins,scaffold proteins,metabolism enzymes and some fertility promoted proteins,coupled with the higher levels of protein phosphorylation,acetylation and succinylation in Meishan boar sperm,may impaired sperm motility and energy metabolism,could be one of the main reasons for explaining the higher quality and reproductive ability of sperm in Meishan than Duroc boar. |
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