Abstract
Background: Boar seminal plasma is rich in cytokines, which could influence the capability of spermatozoa to tolerate preservation.Objectives: To evaluate the involvement of boar seminal plasma cytokines in the changes experienced by boar spermatozoa during their storage, either in liquid or frozen state.Materials and Methods: In two separated experiments, semen samples from healthy and fertile boars were split in two aliquots, one centrifuged twice (1,500 ×g for 10 min) to harvest seminal plasma, whereas the other was either commercially extended (3 × 107 sperm/mL) and liquid-stored at 17°C during 144 h (n = 28, Experiment 1) or frozen-thawed using a standard 0.5 mL protocol (n = 27, Experiment 2). Sixteen cytokines were quantified using Luminex xMAP®. Sperm attributes (CASA-evaluated total and progressive motility; flow cytometry-evaluated sperm viability, production of intracellular H2O2 and and levels of lipid peroxidation in viable spermatozoa) were evaluated either at 0, 72, or 144 h of liquid storage (Experiment 1) or before freezing and at 30- and 150-min post-thawing (Experiment 2).Results: Multiple linear regression models, with Bayesian approach for variable selection, revealed that the anti-inflammatory TGF-β2, TGF-β3, IL-1Ra, and IL-4 and the pro-inflammatory IL-8 and IL-18, predicted changes in sperm motility for liquid-stored semen while the anti-inflammatory IFN-γ was included in the models predicting changes in all sperm attributes for cryopreserved semen.Conclusion: Specific boar seminal plasma cytokines would contribute to modulate the structural and metabolic changes shown by spermatozoa during preservation, either in liquid or frozen state.
Highlights
Cytokines comprise a family of ubiquitous low-molecular weight proteins released mainly by immune cells, mediating intercellular regulation of multiple cellular functions [1]
A repertoire of cytokines has been identified in Seminal Cytokines in Semen Preservation seminal plasma (SP) of humans [7], and other several mammalian species [8,9,10,11], including pigs, with quantitative variation among ejaculates [12]
Recent findings, indicating that boar SP is rich in cytokines but with concentrations differing substantially between mature boars routinely used in artificial insemination (AI) programs [12], provides the relevant pre-requisite to evaluate whether these SP-cytokines would affect the capacity of boar spermatozoa to sustain preservation, either at liquid- or at frozen state
Summary
Cytokines comprise a family of ubiquitous low-molecular weight proteins released mainly by immune cells, mediating intercellular regulation of multiple cellular functions [1]. A repertoire of cytokines has been identified in Seminal Cytokines in Semen Preservation seminal plasma (SP) of humans [7], and other several mammalian species [8,9,10,11], including pigs, with quantitative variation among ejaculates [12]. Recent findings, indicating that boar SP is rich in cytokines but with concentrations differing substantially between mature boars routinely used in AI programs [12], provides the relevant pre-requisite to evaluate whether these SP-cytokines would affect the capacity of boar spermatozoa to sustain preservation, either at liquid- or at frozen state. Boar seminal plasma is rich in cytokines, which could influence the capability of spermatozoa to tolerate preservation
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