Concentration, activity and biochemical characterization of myeloperoxidase in fresh and post-thaw equine semen and their implication on freezability.
Abstract: Myeloperoxidase (MPO) is a pro-oxidant enzyme associated with decreased motility in thawed equine semen. This study aimed to describe MPO concentration, activity and subunits in raw and thawed semen and to correlate these data with motilities in raw and thawed semen. Semen samples from five stallions were collected four times. Motilities were assessed in raw and thawed semen. MPO assays were performed in raw seminal plasma, raw sperm-rich pellet and thawed semen. Total and active MPO concentrations were, respectively, assayed by enzyme-linked immunosorbent assay and specific immunological extraction followed by enzymatic detection. MPO subunits present in semen were characterized by Western blot. Purified active MPO was added in saline solution and freezing extender to control its activity during freezing procedure. Differences between medians were determined using Kruskal-Wallis test, and correlations were determined using Spearman's test for nonparametric data. Active MPO concentration was low in seminal plasma and thawed semen, but high in pellet (p = 0.0058), as the opposite relation was observed for total MPO concentration (p < 0.0001). In seminal plasma and post-thaw semen, inactive 86-kDa MPO precursor was mainly observed. Purified MPO activity was decreased in the extender (p = 0.0286). MPO activity in pellet was highly correlated with thawed progressive motility (r = -0.5576, p = 0.0086). Inactive MPO precursor and unknown low molecular weight inactive MPO precursor subunits explain low MPO activity in semen. Major MPO activity was observed in pellet, and post-thaw loss of activity is partially explained by MPO inactivation in extender. Thawed semen motility was negatively correlated with MPO activity in pellet, becoming a potential freezability predictor.
© 2014 Blackwell Verlag GmbH.
Publication Date: 2014-01-31 PubMed ID: 24479950DOI: 10.1111/rda.12270Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study seeks to understand the role of myeloperoxidase (MPO), a pro-oxidant enzyme, in the movement of thawed horse semen. The research examines the MPO amount, activity, and structure in fresh and thawed semen, and its correlation with the motility of the semen. Results indicate that MPO activity impacts the motility of thawed semen and could potentially predict the quality of thawed semen.
Semen Sample Collection and Assessment
- Semen samples were collected from five stallions, and this process was performed four times.
- The movement of the semen cells was assessed both in their natural state and after thawing.
- The research carried out MPO tests in raw seminal plasma, a dense fraction of sperm collected from semen, and in thawed semen.
MPO Analysis
- The total and active MPO concentrations were assessed using different biochemical tests.
- MPO structures present in semen were also characterized using a method known as Western blot.
- Active MPO was added in saline solution and freezing extender to check its activity during the freezing process.
- Statistical analysis was performed for differences between median values and to establish correlations.
Findings and Conclusions
- Active MPO concentration was found to be low in seminal plasma and thawed semen but high in the semen-rich pellet.
- The inactive MPO precursor was mainly observed in seminal plasma and post-thaw semen; this and other inactive MPO precursor subunits justify the low MPO activity in semen.
- Semen motility after thawing was negatively correlated with MPO activity in the semen-rich pellet. This indicates that higher MPO activity leads to a decrease in semen motility after thawing.
- Significant MPO activity was observed in the semen-rich pellet, and post-thaw loss of activity is partially explained by MPO inactivation in the freezing extender.
- These findings suggest that MPO activity in the semen-pellet has potential as a predictor of how well semen can withstand the freezing process.
Cite This Article
APA
Ponthier J, Franck T, Parrilla-Hernandez S, Niesten A, de la Rebiere G, Serteyn D, Deleuze S.
(2014).
Concentration, activity and biochemical characterization of myeloperoxidase in fresh and post-thaw equine semen and their implication on freezability.
Reprod Domest Anim, 49(2), 285-291.
https://doi.org/10.1111/rda.12270 Publication
Researcher Affiliations
- Equine Clinic, Veterinary Medicine Faculty, ULg University of Liège, Liège, Belgium; LINALUX-MLS, Centre Européen du Cheval, Vielsalm, Belgium.
MeSH Terms
- Animals
- Cryopreservation / veterinary
- Gene Expression Regulation, Enzymologic / physiology
- Horses / physiology
- Male
- Peroxidase / genetics
- Peroxidase / metabolism
- Semen / enzymology
- Semen Preservation / veterinary
Citations
This article has been cited 2 times.- Parrilla Hernández S, Franck T, Munaut C, Feyereisen É, Piret J, Farnir F, Reigner F, Barrière P, Deleuze S. Characterization of Myeloperoxidase in the Healthy Equine Endometrium. Animals (Basel) 2023 Jan 21;13(3).
- Egyptien S, Deleuze S, Ledeck J, Ponthier J. Sperm Quality Assessment in Stallions: How to Choose Relevant Assays to Answer Clinical Questions. Animals (Basel) 2023 Oct 6;13(19).
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