[en] Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in, and released by, neutrophils during degranulation or after lysis. Post-thaw semen contains MPO and its concentration is associated with decreased sperm motility. Recently, MPO concentration in post-thaw semen was shown to be associated with presence of non-sperm cells. The aim of this study was to describe the effect of centrifugal fractionation of semen prior to cryopreservation on post-thaw concentrations of non-sperm cells and MPO.
The experimental design consisted in freezing semen with or without previous centrifugation through two concentrations of single layer colloid media. Non-sperm cells and MPO concentrations were assessed in pellet and upper layer at each step of the procedure and MPO was detected in cells by immunocytochemistry.
Single layer colloid centrifugation decreased non-sperm cells and MPO concentrations in post-thaw semen. The MPO concentration was correlated with concentration of non-sperm cells in the upper layer of the supernatant. In post-thaw semen, with or without previous single layer colloid centrifugation, MPO concentration was correlated with concentration of non-sperm cells. Overall, neutrophils were rarely observed and non-sperm cells were mainly epithelial cells or cellular debris, as demonstrated by MPO immunocytochemistry.
Following single layer colloid centrifugation, MPO concentration was decreased, and correlations observed in different samples of the experiments revealed an association between concentrations of MPO and non-sperm cells. At all steps of the semen processing and cryopreservation, MPO immunostaining was clearly identified only on non-sperm cells. Our results demonstrate that non-sperm cells present in fresh semen release MPO during freezing.
Research center :
Clinique Equine, Département des sciences cliniques des animaux de compagnie et équidés, Faculté de Médecine Vétéri Equine theriogenology, Large Animal Clinical Sciences Department, Texas A&M university
Disciplines :
Animal production & animal husbandry Veterinary medicine & animal health Biotechnology
Author, co-author :
Ponthier, Jérôme ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Obstétrique et pathologie de la reproduction des équidés
Teague, Sheila; Texas A&M university > Departement of Large Animal Clinical Sciences > Equine Theriogenology
Franck, Thierry ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
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
Brinsko, Steven; Texas A&M university > Departement of Large Animal Clinical Sciences > Equine theriogenology
Love, Charles; Texas A&M university > Departement of Large Animal Clinical Sciences > Equine theriogenology
Blanchard, Terry; Texas A&M university > Departement of Large Animal Clinical Sciences > Equine theriogenology
Varner, Dickson; Texas A&M university > Departement of Large Animal Clinical Sciences > Equine theriogenology
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 :
Effect of non-sperm cells removal with single layer colloidal centrifugation on myeloperoxidase concentration in post-thaw equine semen
Publication date :
December 2013
Journal title :
Theriogenology
ISSN :
0093-691X
eISSN :
1879-3231
Publisher :
Elsevier, New York, United States - New York
Volume :
80
Issue :
9
Pages :
1082-1087
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
SPERMPO
Funders :
ULiège - Université de Liège [BE] Texas A&M university Clinique Equine, Département des sciences cliniques des animaux de compagnie et équidés, Faculté de Médecine Vétérinaire, Ulg
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