Proteomic identification of boar seminal plasma proteins related to sperm resistance to cooling at 17 °C

Lazari, Franciele Lucca de; Sontag, Elistone Rafael; Schneider, Alexander; Moura, Arlindo A.; Vasconcelos, Fábio Perdigão; Nagano, Celso Shiniti; Dalberto, Pedro Ferrari; Bizarro, Cristiano Valim; Mattos, Rodrigo Costa; Jobim, Mariana; Bustamante‐Filho, Ivan Cunha

doi:10.1016/j.theriogenology.2019.11.023

Cited by 17 publications

(20 citation statements)

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“…The application of this emerging approach for the identification of novel SP quality and fertility biomarkers is currently flourishing [9]. In this regard, great efforts have been devoted to exploring new biomarkers in pig SP through comparative proteomics [10][11][12][13]. Specifically, recent studies uncovered the role of antioxidant enzymes in SP, such as glutathione peroxidase 5 (GPX5) and paraoxonase 1 (PON1), as sperm cryotolerance, quality and/or fertility biomarkers [14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Exploring Seminal Plasma GSTM3 as a Quality and In Vivo Fertility Biomarker in Pigs—Relationship with Sperm Morphology

et al. 2020

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Glutathione S-transferases Mu 3 (GSTM3) is an essential antioxidant enzyme whose presence in sperm has recently been related to sperm cryotolerance, quality and fertility. However, its role in seminal plasma (SP) as a predictor of the same sperm parameters has never been investigated. Herein, cell biology and proteomic approaches were performed to explore the presence, origin and role of SP-GSTM3 as a sperm quality and in vivo fertility biomarker. GSTM3 in SP was quantified using a commercial Enzyme-Linked Immunosorbent Assay (ELISA) kit specific for Sus scrofa, whereas the presence of GSTM3 in testis, epididymis and accessory sex glands was assessed through immunoblotting analysis. Sperm quality and functionality parameters were evaluated in semen samples at 0 and 72 h of liquid-storage, whereas fertility parameters were recorded over a 12-months as farrowing rate and litter size. The presence and concentration of GSTM3 in SP was established for the first time in mammalian species, predominantly synthesized in the epididymis. The present study also evidenced a relationship between SP-GSTM3 and sperm morphology and suggested it is involved in epididymal maturation rather than in ejaculated sperm physiology. Finally, the data reported herein ruled out the role of this antioxidant enzyme as a quality and in vivo fertility biomarker of pig sperm.

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Section: Introductionmentioning

confidence: 99%

Exploring Seminal Plasma GSTM3 as a Quality and In Vivo Fertility Biomarker in Pigs—Relationship with Sperm Morphology

et al. 2020

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“…Also, it contributes to proteolytic networks as an activator of downstream serine proteases and metalloproteases frequently involved tumorigenesis and invasion 48,49 . Recent studies have demonstrated that cathepsin B is overexpressed in the seminal plasma of boars with low seminal quality 45 and boars with high resistance to storage at 17°C 10 . Two possible events could explain these contrasting findings: (a) the intrinsic enzymatic activity of cathepsin B, dependent on maintenance of its native state, and (b) the presence of cathepsin B inhibitors.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, the use of seminal biomarkers associated with seminal resistance to cooling could bring several benefits to boar stud farms. Recently, the association of individual boar sperm resistance to storage at 17°C for up to 144 hours with proteins of the seminal plasma has been already established 10 . However, it remains unclear whether seminal plasma proteins would modulate the resistance to chilling injury at 5°C.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, the association of individual boar sperm resistance to storage at 17°C for up to 144 hours with proteins of the seminal plasma has been already established. 10 However, it remains unclear whether seminal plasma proteins would modulate the resistance to chilling injury at 5°C. Thus, the present study aimed to compare seminal plasma proteomes of boars with high and low resistance to hypothermic preservation at 5°C, identifying putative biomarkers for seminal resistance to cooling.…”

Section: Introductionmentioning

confidence: 99%

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Differential seminal plasma proteome signatures of boars with high and low resistance to hypothermic semen preservation at 5°C

et al. 2020

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Background Hypothermic storage at 5°C has been investigated as an alternative to promote the prudent use of antibiotics for boar artificial insemination doses. However, this temperature is challenging for some ejaculates or boars. Objective The present study aimed to identify putative biomarkers for semen resistance to hypothermic storage at 5°C by comparing the seminal plasma proteomes of boars with high and low seminal resistance to preservation at 5°C. Materials and methods From an initial group of 34 boars, 15 were selected based on the following criteria: ejaculate with ≤20% abnormal spermatozoa and at least 70% progressive motility at 120 hours of storage at 17°C. Then, based on the response to semen hypothermic storage at 5°C, boars were classified into two categories: high resistance—progressive motility of >75% in the three collections (n = 3); and low resistance—progressive motility of <75% in the three collections (n = 3). Seminal plasma proteins were analyzed in pools, and differential proteomics was performed using Multidimensional Protein Identification Technology. Results Progressive motility was lower at 120 hours of storage in low resistance, compared to high resistance boars (P < .05). Acrosome and plasma membrane integrity were not affected by the boar category, storage time, or their interaction (P ≥ .104). Sixty‐five proteins were considered for differential proteomics. Among the differentially expressed and exclusive proteins, the identification of proteins such cathepsin B, legumain, and cystatin B suggests significant changes in key enzymes (eg, metalloproteinases) involved in spermatogenesis, sperm integrity, and fertilizing potential. Discussion and Conclusion Differences in the seminal plasma suggest that proteins involved in the proteolytic activation of metalloproteinases and proteins related to immune response modulation could disrupt key cellular pathways during spermatogenesis and epididymal maturation, resulting in altered resistance to chilling injury. Further in vivo studies focusing on the immunological crosstalk between epithelial cells and gametes might explain how the immune regulators influence sperm resistance to hipothermic storage.

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“…Moreover, boar sperm longevity and survivability varies among individuals [8]. Despite the optimal quality assessed before cooling, ejaculates may respond differently to liquid preservation [9]. Predicting the fertility outcome of cooled or frozen spermatozoa is one of the most relevant aspects in the field of porcine reproduction [5].…”

Section: Introductionmentioning

confidence: 99%

Proteomic Analysis of Fresh and Liquid-Stored Boar Spermatozoa

Bajuk

Zrimšek

Pipan

et al. 2020

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In this study comparative proteomics was used to define changes in the expression of the spermatozoa proteins during liquid storage. Semen from eight boars was analyzed on the day of collection and after liquid preservation at 15–17 °C for three days. Sperm parameters (concentration, motility, morphology, vitality) and percentage of non-capacitated and acrosomal-reacted spermatozoa were determined. Sperm proteins were extracted and separated by two-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and proteomic profiles were computationally compared to highlight differentially expressed protein spots that were, in turn, identified by mass spectrometry. The intensities of four spots were significantly different between fresh and liquid stored sperm. Namely: ATP citrate lyase, chaperonin containing T-complex polypeptide 1 (TCP1) subunit ε and probable phospholipid-transporting ATP-ase were over-expressed in liquid stored sperm, whereas cytosolic non-specific dipeptidase was over-expressed in fresh sperm. These differentially expressed proteins could be used as plausible biomarkers for the evaluation of boar semen quality and spermatozoa survival after liquid storage and could help to address problems associated with sperm preservation.

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Proteomic identification of boar seminal plasma proteins related to sperm resistance to cooling at 17 °C

Cited by 17 publications

References 73 publications

Exploring Seminal Plasma GSTM3 as a Quality and In Vivo Fertility Biomarker in Pigs—Relationship with Sperm Morphology

Exploring Seminal Plasma GSTM3 as a Quality and In Vivo Fertility Biomarker in Pigs—Relationship with Sperm Morphology

Differential seminal plasma proteome signatures of boars with high and low resistance to hypothermic semen preservation at 5°C

Proteomic Analysis of Fresh and Liquid-Stored Boar Spermatozoa

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