[en] 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
3 blot. Purified active MPO was added in PBS 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). Inac- tive 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 inactiva- tion in extender. Thawed semen motility was negatively correlated with MPO activity in pellet, becoming a potential freezability predictor.
Research Center/Unit :
ULiège - FMV - Clinique Equine LINALUX-MLS Centre Européen du Cheval ULiège - CORD
Disciplines :
Biotechnology Animal production & animal husbandry Veterinary medicine & animal health
Author, co-author :
Ponthier, Jérôme ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Franck, Thierry ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Parrilla Hernandez, Sonia ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Département clinique des animaux de compagnie et des équidés
Niesten, Ariane ; Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
de la Rebière de Pouyade, Geoffroy ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Serteyn, Didier ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Deleuze, Stefan ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Obstét. et path. de la reprod. des anim. de comp. et équidés
Language :
English
Title :
Concentration, Activity and Biochemical Characterization of Myeloperoxidase in Fresh and Post-thaw equine semen and their Implication on Freezability
Publication date :
2014
Journal title :
Reproduction in Domestic Animals
ISSN :
0936-6768
eISSN :
1439-0531
Publisher :
Blackwell Publishing, Berlin, Germany
Volume :
49
Pages :
285–291
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
SPERMPO
Funders :
ULg - FMV - Clinique Equine LINALUX-MLS Centre Européen du Cheval ULg - CORD
Agarwal A, Sharma RK, Nallella KP, Thomas AJ Jr, Alvarez JG, Sikka SC, 2006: Reactive oxygen species as an independent marker of male factor infertility. Fertil Steril 86, 878-885.
Aitken RJ, 1995: Free radicals, lipid peroxidation and sperm function. Reprod Fertil Dev 7, 659-668.
Art T, Franck T, Lekeux P, de Moffarts B, Couetil L, Becker M, Kohnen S, Deby-Dupont G, Serteyn D, 2006: Myeloperoxidase concentration in bronchoalveolar lavage fluid from healthy horses and those with recurrent airway obstruction. Can J Vet Res 70, 291-296.
Baumber J, Ball BA, Gravance CG, Medina V, Davies-Morel MC, 2000: The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation. J Androl 21, 895-902.
Baumber J, Vo A, Sabeur K, Ball BA, 2002: Generation of reactive oxygen species by equine neutrophils and their effect on motility of equine spermatozoa. Theriogenology 57, 1025-1033.
Edwards SW, Nurcombe HL, Hart CA, 1987: Oxidative inactivation of myeloperoxidase released from human neutrophils. Biochem J 245, 925-928.
Franck T, Grulke S, Deby-Dupont G, Deby C, Duvivier H, Peters F, Serteyn D, 2005: Development of an enzyme-linked immunosorbent assay for specific equine neutrophil myeloperoxidase measurement in blood. J Vet Diagn Invest 17, 412-419.
Franck T, Kohnen S, Deby-Dupont G, Grulke S, Deby C, Serteyn D, 2006: A specific method for measurement of equine active myeloperoxidase in biological samples and in in vitro tests. J Vet Diagn Invest 18, 326-334.
Franck T, Kohnen S, Boudjeltia KZ, Van Antwerpen P, Bosseloir A, Niesten A, Gach O, Nys M, Deby-Dupont G, Serteyn D, 2009: A new easy method for specific measurement of active myeloperoxidase in human biological fluids and tissue extracts. Talanta 80, 723-729.
Franck T, Votion DM, Ceusters J, La Rebiere D, de Pouyade G, Mouithys-Mickalad A, Niesten A, Fraipont A, Van Erck E, Goachet AG, Robert C, Serteyn D, 2010: Specific immuno-extraction followed by enzymatic detection (SIEFED) of myeloperoxidase and mitochondrial complex I in muscular microbiopsies: preliminary results in endurance horses. Equine Vet J 42(Suppl 38), 296-302.
Graham-Pole J, Davie M, Willoughby ML, 1977: Cryopreservation of human granulocytes in liquid nitrogen. J Clin Pathol 30, 758-762.
Grulke S, Franck T, Gangl M, Peters F, Salciccia A, Deby-Dupont G, Serteyn D, 2008: Myeloperoxidase assay in plasma and peritoneal fluid of horses with gastrointestinal disease. Can J Vet Res 72, 37-42.
Henkel R, Kierspel E, Stalf T, Mehnert C, Menkveld R, Tinneberg HR, Schill WB, Kruger TF, 2005: Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients. Fertil Steril 83, 635-642.
Johansson CS, Matsson FC, Lehn-Jensen H, Nielsen JM, Petersen MM, 2008: Equine spermatozoa viability comparing the NucleoCounter SP-100 and the eosin-nigrosin stain. Anim Reprod Sci 107, 325-326.
Mathy-Hartert M, Bourgeois E, Grulke S, Deby-Dupont G, Caudron I, Deby C, Lamy M, Serteyn D, 1998: Purification of myeloperoxidase from equine polymorphonuclear leucocytes. Can J Vet Res 62, 127-132.
Mazzilli F, Rossi T, Sabatini L, Pulcinelli FM, Rapone S, Dondero F, Gazzaniga PP, 1995: Human sperm cryopreservation and reactive oxygen species (ROS) production. Acta Eur Fertil 26, 145-148.
Meseguer M, Garrido N, Simon C, Pellicer A, Remohi J, 2004: Concentration of glutathione and expression of glutathione peroxidases 1 and 4 in fresh sperm provide a forecast of the outcome of cryopreservation of human spermatozoa. J Androl 25, 773-780.
O'Flaherty CM, Beorlegui NB, Beconi MT, 1999: Reactive oxygen species requirements for bovine sperm capacitation and acrosome reaction. Theriogenology 52, 289-301.
Ortega-Ferrusola C, Garcia BM, Gallardo-Bolanos JM, Gonzalez-Fernandez L, Rodriguez-Martinez H, Tapia JA, Pena FJ, 2009: Apoptotic markers can be used to forecast the freezeability of stallion spermatozoa. Anim Reprod Sci 114, 393-403.
Parrilla-Hernandez S, Deleuze S, Van Den Berghe F, Ponthier J, 2012: Raw semen concentration directly influences CASA velocity pathways. J Equine Vet Sci 32, 505.
Ponthier J, de la Rebiere de Pouyade GDesvals M, Spalart M, Franck T, Serteyn D, Deleuze S, 2010a: Is neutrophil elastase associated with myeloperoxidase concentration and post-thawing parameters in equine frozen semen? Anim Reprod Sci 121S, S200-S202.
Ponthier J, Franck T, Detilleux J, Mottart E, Serteyn D, Deleuze S, 2010b: Association between Myeloperoxidase Concentration in Equine Frozen Semen and Post-Thawing Parameters. Reprod Domest Anim 45, 811-816.
Ponthier J, Desvals M, Franck T, de la Rebiere de Pouyade G, Spalart M, Palmer E, Serteyn D, Deleuze S, 2012: Myeloperoxidase in equine semen: concentration and Localization during freezing processing. J Equine Vet Sci 32, 32-37.
Ponthier J, Teague SR, Franck T, de la Rebiere de Pouyade G, Serteyn D, Brinsko SP, Love CC, Blanchard TL, Varner DD, Deleuze S, 2013: Effect of non-sperm cells removal with single-layer colloidal centrifugation on Myeloperoxidase concentration in post-thaw equine semen. Theriogenology 80, 1082-1087.
de la Rebiere G, Riggs LM, Moore JN, Franck T, Deby-Dupont G, Hurley DJ, Serteyn D, 2010: Equine neutrophil elastase in plasma, laminar tissue, and skin of horses administered black walnut heartwood extract. Vet Immunol Immunopathol 135, 181-187.
Roumeguere T, Delree P, Van Antwerpen P, Rorive S, Vanhamme L, de Ryhove LD, Serteyn D, Wespes E, Vanhaerverbeek M, Boudjeltia KZ, 2012: Intriguing location of myeloperoxidase in the prostate: a preliminary immunohistochemical study. Prostate 72, 507-513.
Serteyn D, Grulke S, Franck T, Mouithys-Mickalad A, Deby-Dupont G, 2003: La myeloperoxydase des neutrophiles, une enzyme de défense au capacité oxydantes. Neutrophil myeloperoxidase, an oxidant defensive enzyme. Ann Méd Vét 147, 79-93.
Sion B, Janny L, Boucher D, Grizard G, 2004: Annexin V binding to plasma membrane predicts the quality of human cryopreserved spermatozoa. Int J Androl 27, 108-114.
Taurog A, Dorris ML, 1992: Myeloperoxidase-catalyzed iodination and coupling. Arch Biochem Biophys 296, 239-246.
Wang A, Fanning L, Anderson DJ, Loughlin KR, 1997a: Generation of reactive oxygen species by leukocytes and sperm following exposure to urogenital tract infection. Arch Androl 39, 11-17.
Wang AW, Zhang H, Ikemoto I, Anderson DJ, Loughlin KR, 1997b: Reactive oxygen species generation by seminal cells during cryopreservation. Urology 49, 921-925.
Wang Y, Sharma RK, Agarwal A, 1997c: Effect of cryopreservation and sperm concentration on lipid peroxidation in human semen. Urology 50, 409-413.
