Toxicity of glycerol for the stallion spermatozoa: Effects on membrane integrity and cytoskeleton, lipid peroxidation and mitochondrial membrane potential

Funding Agencies|Ministerio de Ciencia e Innovacion-FEDER Madrid, Spain|AGL 2010 20758 (GAN)|Inia|RZ2008-00018-00-00|Junta de Extremadura FEDER GR|10010|
Glycerol is, to date, the most widely used cryoprotectant to freeze stallion spermatozoa at concentrations between 2% and 5%. Cryoprotectant toxicity has been claimed to be the single most limiting factor for the success of cryopreservation. In order to evaluate the toxic effects of the concentrations of glycerol used in practice, stallion spermatozoa were incubated in Biggers Whitten and Whittingham (BWW) media supplemented with 0%, 0.5%, 1.5%, 2.5%, 3.5%, and 5% glycerol. In two additional experiments, a hyposmotic (75 mOsm/kg) and a hyperosmotic (900 mOsm/kg) control media were included. Sperm parameters evaluated included cell volume, membrane integrity, lipid peroxidation, caspase 3, 7, and 8 activation, mitochondrial membrane potential, and integrity of the cytoskeleton. Glycerol exerted toxicity at concentrations 3.5% and the maximal toxicity was observed at 5%. The actin cytoskeleton was especially sensitive to glycerol presence, inducing rapid F actin depolymerization at concentrations over 1.5%. The sperm membrane and the mitochondria were other structures affected. The toxicity of glycerol is apparently related to osmotic and nonosmotic effects. In view of our results the concentration of glycerol in the freezing media for stallion spermatozoa should not surpass 2.5%.

Ortega Ferrusola, CUniversity of Extremadura (författare)
Aparicio, I MUniversity of Extremadura (författare)
Miro-Moran, AUniversity of Extremadura (författare)
Morillo Rodriguez, AUniversity of Extremadura (författare)
Gallardo Bolanos, J MUniversity of Extremadura (författare)
Gonzalez Fernandez, LUniversity of Extremadura (författare)
Balao da Silva, C MUniversity of Extremadura (författare)
Rodriguez-Martinez, HeribertoLinköpings universitet,Institutionen för klinisk och experimentell medicin,Hälsouniversitetet(Swepub:liu)herro55 (författare)
Tapia, J AUniversity of Extremadura (författare)
Pena, F JUniversity of Extremadura (författare)
University of ExtremaduraInstitutionen för klinisk och experimentell medicin (creator_code:org_t)

Av författaren/redakt...: Macias Garcia, B; Ortega Ferrusola ...; Aparicio, I M; Miro-Moran, A; Morillo Rodrigue ...; Gallardo Bolanos ...; visa fler...; Gonzalez Fernand ...; Balao da Silva, ...; Rodriguez-Martin ...; Tapia, J A; Pena, F J; visa färre...

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