Abstract
BackgroundMature spermatozoa contain numerous epididymal and seminal plasma proteins, which full identification through high-throughput technologies may allow for a better understanding of the sperm biology. Therefore, we conducted a global proteomic analysis of boar spermatozoa through shotgun and gel-based methodologies.ResultsThe total proteins were extracted from mature spermatozoa and subjecsted to proteome analyses. Functional analyses of gene ontology representations and pathway enrichments were conducted on the shotgun dataset, followed by immunology and gene expression validations. Shotgun and gel-based approaches allowed the detection of 2728 proteins and 2123 spots, respectively. Approximately 38% and 59% of total proteins were respectively fully and partially annotated, and 3% were unknown. Gene ontology analysis indicated high proportions of proteins associated with intracellular and cytoplasm localizations, protein and nucleic acid binding, hydrolase and transferase activities, and cellular, metabolic, and regulation of biological processes. Proteins associated with phosphorylation processes and mitochondrial membranes, nucleic acid binding, and phosphate and phosphorous metabolics represented 77% of the dataset. Pathways associated with oxidative phosphorylation, citrate cycle, and extra-cellular matrix-receptor interaction were significantly enriched. Protein complex, intracellular organelle, cytoskeletal parts, fertilization and reproduction, and gap junction pathway were significantly enriched within the top 116 highly abundant proteins. Nine randomly selected protein candidates were confirmed with gel-based identification, immunofluorescence detection, and mRNA expression.ConclusionsThis study offers an in-depth proteomic mapping of mature boar spermatozoa that will enable comparative and discovery research for the improvement of male fertility.
Highlights
Mature spermatozoa contain numerous epididymal and seminal plasma proteins, which full identification through high-throughput technologies may allow for a better understanding of the sperm biology
Each semen ejaculate was extended with the Beltsville Thawing Solution (BTS: Minitube of America – MOFA; Verona, WI) at the boar stud facilities and overnight-shipped to our laboratory, in three different occasions
Total of 2406 detected proteins were annotated with NCBI-nr databases, corresponding to 781 (29%) full and 1625 (59%) partial annotations
Summary
Mature spermatozoa contain numerous epididymal and seminal plasma proteins, which full identification through high-throughput technologies may allow for a better understanding of the sperm biology. Spermatozoa are haploid cells produced in the testis with a specific shape allowing them to carry and deliver paternal materials to the oocyte Their biology has a multifaceted array of genetic, proteomic and metabolic differences, which unpredictable interactions with one another could influence the sperm function and male fertility. Spermatozoa are released within the seminiferous tubules upon their production and maintain continuous interactions with surrounding secretions of the genital tract (testes and epididymis) throughout their longitudinal migration within the Feugang et al BMC Genomics (2018) 19:62 epididymis lumen and accessory sex glands (prostate, cowper’s gland and seminal vesicles) These proteinbased interactions contribute to shaping the sperm structure and influence their fertility [10,11,12,13,14].
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