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Myeloperoxidase activity decreases in equine semen freezing extenders
Ponthier, Jérôme; Franck, Thierry; Niesten, Ariane et al.
2013In Amir, Arav (Ed.) Proceedings of the 3rd Cryo Congress
Peer reviewed
 

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Keywords :
Semen; Equine; Stallion; Freezing; Cryopreservation; Myeloperoxidase; Reactive oxygen species
Abstract :
[en] Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in and released by neutrophils, and associated with decreased post-thaw motility of equine semen. This study aimed to compare MPO activity in pure equine freezing extender, raw and post-thaw semen. Active MPO Concentration (AMC) was measured with Specific Immunologic Extraction Followed by Enzymatic Detection in 20 ejaculates. Raw semen intra cellular AMC was determined in the supernatant after membrane lysis, each pellet containing 100x106 spermatozoa. AMC was also assayed in supernatants of semen frozen following a conventional method using INRA FreezeTM (IMV, France). Effect of freezing procedure on AMC was tested by experimentally adding 500ng of purified active MPO (Calbiochem, Germany) in 4 samples either with 5ml of PBS or INRA FreezeTM before assay. AMC was higher in sperm-rich pellet (0.306ng/ml) than in post-thaw semen (0.002ng/ml) (p=0.058). After experimental MPO addition, no activity variation was observed during the freezing procedure (after dilution, 1, 2 hours of cooling and post-thawing) within the same medium. Purified MPO activity was decreased in INRA FreezeTM when compared to PBS at all timings of sampling (p=0.0286). When all samples were pooled, remaining activity in INRA FreezeTM was 23.93±13.13%. MPO fixation on large proteins contained in the extender experimentally reduces AMC, as previously observed in plasma. However, AMC decrease observed during semen freezing is more important than after experimental addition. That could be explained by a MPO interaction with seminal plasma, a partial MPO release or a MPO inactivation during equine semen freezing.
Research center :
ULiège - FMV - Obstétrique et pathologie de la reproduction des équidés
ULiège - Center for Oxygen Research and Development (CORD)
Centre Européen du cheval - LINALUX-Mont le Soie
Disciplines :
Veterinary medicine & animal health
Animal production & animal husbandry
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
Niesten, Ariane ;  Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
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
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 :
Myeloperoxidase activity decreases in equine semen freezing extenders
Publication date :
23 March 2013
Event name :
3rd international congress on controversies in cryopreservation of stem cells, reproductive cells tissue and organs (CRYO 3)
Event place :
Berlin, Germany
Event date :
21 - 23 mars 2013
Audience :
International
Main work title :
Proceedings of the 3rd Cryo Congress
Editor :
Amir, Arav
Publisher :
Cemtecmed
Pages :
A-2
Peer reviewed :
Peer reviewed
Name of the research project :
SPERMPO
Funders :
ULg - FMV - Obstétrique et pathologie de la reproduction des équidés
ULg - Center for Oxygen Research and Development (CORD)
EquiDir - Centre Européen du cheval - Linalux-Mont le Soie [BE]
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since 29 April 2013

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