Effect of several antioxidants on thawed ram spermatozoa submitted to 37ºC up to four hours

Thawed ram spermatozoa were incubated at 37 ◦C in presence of dehydroascorbic acid (DHA), TEMPOL (TPL), N-acetyl-cysteine (NAC) and rutin (RUT), at 0.1 and 1 mM, in order to test their effects on sperm physiology. Cryopreserved spermatozoa from four rams were thawed, pooled, washed and incubated in...

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Detalles Bibliográficos
Autores: Mata Campuzano, María, Álvarez García, Mercedes, Álvarez Rodríguez, Manuel, Anel López, Luis, Paz, Paulino de, Garde López-Brea, José Julián, Martínez Pastor, Felipe
Tipo de recurso: artículo
Fecha de publicación:2012
País:España
Institución:Universidad de Castilla-La Mancha
Repositorio:RUIdeRA. Repositorio Institucional de la UCLM
OAI Identifier:oai:ruidera.uclm.es:10578/7474
Acceso en línea:http://hdl.handle.net/10578/7474
Access Level:acceso abierto
Palabra clave:Antioxidantes
Estrés oxidativo
Ganado ovino
Espermatozoides
Vitamina C
Descripción
Sumario:Thawed ram spermatozoa were incubated at 37 ◦C in presence of dehydroascorbic acid (DHA), TEMPOL (TPL), N-acetyl-cysteine (NAC) and rutin (RUT), at 0.1 and 1 mM, in order to test their effects on sperm physiology. Cryopreserved spermatozoa from four rams were thawed, pooled, washed and incubated in TALP-Hepes with 1 mM or 0.1 mM of each antioxidant, performing a replicate with induced oxidative stress e2+/ascorbate). Motility (CASA), viability and mitochondrial membrane potential (flow cytometry) were analyzed at 2 and 4 h. Lipoperoxidation (MDA production), intracellular ROS and DNA status (TUNEL) were analyzed at 4 h. Antioxidants, except DHA 0.1 mM, decreased motility and kinematic parameters, but had little effect on viability or mitochondrial activity. Except 1 mM DHA, the antioxidants reduced ROS at 4 h. Moreover, NAC 1 mM, rutin and TEMPOL reduced ROS and DNA damage in presence of oxidative stress. NAC, rutin 1 mM and TEMPOL reduced lipoperoxidation in presence of oxidative stress. However, DHA did not affected lipoperoxidation. At 1 mM, DHA increased DNA damage in absence of oxidative stress. DHA effects could arise from spermatozoa having a low capacity for reducing it to ascorbic acid, and it may be tested in presence of other antioxidants or reducing power. Future research should focus in testing if the inhibition of motility observed for NAC, rutin and TEMPOL is reversible. These antioxidants might be useful at lower temperatures (refrigerated storage or cryopreservation) when their protective effects could be advantageous