Comparative Proteomics and Phosphoproteomics Analysis Reveal the Possible Breed Difference in Yorkshire and Duroc Boar Spermatozoa

Abstract

Sperm cells are of unique elongated structure and function, the development of which is tightly regulated by the existing proteins and the posttranslational modifications (PTM) of these proteins. Based on the phylogenetic relationships of various swine breeds, Yorkshire boar is believed to be distinctly different from Duroc boar. The comprehensive differential proteomics and phosphoproteomics profilings were performed on spermatozoa from both Yorkshire and Duroc boars. By both peptide and PTM peptide quantification followed by statistical analyses, 167 differentially expressed proteins were identified from 1,745 proteins, and 283 differentially expressed phosphopeptides corresponding to 102 unique differentially phosphorylated proteins were measured from 1,140 identified phosphopeptides derived from 363 phosphorylated proteins. The representative results were validated by Western blots. Pathway enrichment analyses revealed that majority of differential expression proteins and differential phosphorylation proteins were primarily concerned with spermatogenesis, male gamete generation, sperm motility, energy metabolism, cilium morphogenesis, axonemal dynein complex assembly, sperm–egg recognition, and capacitation. Remarkably, axonemal dynein complex assembly related proteins, such as SMCP, SUN5, ODF1, AKAP3, and AKAP4 that play a key regulatory role in the sperm physiological functions, were significantly higher in Duroc spermatozoa than that of Yorkshire. Furthermore, phosphorylation of sperm-specific proteins, such as CABYR, ROPN1, CALM1, PRKAR2A, and PRKAR1A, participates in regulation of the boar sperm motility mainly through the cAMP/PKA signal pathway in different breeds, demonstrating that protein phosphorylation may be an important mechanism underlying the sperm diversity. Protein–protein interaction analysis revealed that the 14 overlapped proteins between differential expression proteins and differential phosphorylation proteins potentially played a key role in sperm development and motility of the flagellum, including the proteins ODF1, SMCP, AKAP4, FSIP2, and SUN5. Taken together, these physiologically and functionally differentially expressed proteins (DEPs) and differentially expressed phosphorylated proteins (DPPs) may constitute the proteomic backgrounds between the two different boar breeds. The validation will be performed to delineate the roles of these PTM proteins as modulators of Yorkshire and Duroc boar spermatozoa.

Do Motility and Sperm Dose Count Affect In Vivo Fertility in Boar?

“…Differences between breeds in in vivo fertility might be a consequence of certain mechanisms at a molecular level. In this respect, the research of Xu et al [9] suggests multiple changes at the level of proteins and status of phosphorylation between the sperm of Large White and of Duroc breeds, suggesting a correlation between reproductive efficiency and fertilizing potential. They have identified different novel The research of Tremoen et al [8] showed that there are differences between the breeds of Norwegian Landrace and Duroc boars regarding CASA variables and the effect on the total number of piglets at birth, with motility being the most frequently used parameter of sperm quality.…”

“…Differences between breeds in in vivo fertility might be a consequence of certain mechanisms at a molecular level. In this respect, the research of Xu et al [9] suggests multiple changes at the level of proteins and status of phosphorylation between the sperm of Large White and of Duroc breeds, suggesting a correlation between reproductive efficiency and fertilizing potential. They have identified different novel molecular mechanisms that might help to better understand spermatogenesis, spermatozoa motility, energy metabolism and a process of joining spermatozoa and ovum.…”

Quantitative phosphoproteomics analyses reveal the regulatory mechanisms related to frozen-thawed sperm capacitation and acrosome reaction in yak (Bos grunniens)

“…It is worth noting that since sperm are both transcriptionally and translationally silent, their function is highly dependent on the existence of PTMs of the protein complement ( Dun et al, 2012 ; Xu et al, 2021 ). It is necessary for the regulation of sperm function that phosphorylation as the most important PTM can rapidly control the activity of signaling and regulatory proteins ( Parte et al, 2012 ; Dacheux and Dacheux, 2014 ).…”

Should All Fractions of the Boar Ejaculate Be Prepared for Insemination Rather Than Using the Sperm Rich Only?

“…The extender used in our study is recommended for medium-term conservation, which can aid in conserving the sperm quality when different seminal plasma proportions and fraction origin are used. Furthermore, the seminal plasma proteins not only vary between fractions of the ejaculates [ 13 , 14 ], but there are also inter-breed [ 32 ] and inter-male variations [ 33 ]. In our case, no male effect was observed.…”