Kinetics of hydrogen peroxide generated from live and dead ram spermatozoa and the effects of catalase and oxidase substrates addition

https://doi.org/10.17221/8662-CJASCitation:Alomar M., Alzoabi M., Zarkawi M. (2016): Kinetics of hydrogen peroxide generated from live and dead ram spermatozoa and the effects of catalase and oxidase substrates addition. Czech J. Anim. Sci., 61: 1-7.
download PDF
The generation of hydrogen peroxide (H2O2) by ram spermatozoa (spz) was measured using a flurometric assay with 10-acetyl-3,7-dihydroxyphenoxazine agent as a probe for H2O2 detection. The kinetics of H2O2 production from both live and dead spz at 1 × 106, 3 × 106, and6 × 106 spz/well concentrations were assessed in the tyrode albumin lactate (TAL) medium every 15 min for 120 min. An increase in H2O2 production from both live and dead spz was noted with a significant difference (P < 0.05) between the 1 × 106 and 6 × 106 spz/well concentrations. Although dead sperm generated higher amounts of H2O2 than live ones, no significant differences (P > 0.05) were observed between the two types of sperm for the three different concentrations. The generation of H2O2 by ram spz was also compared in the presence and absence of nicotinamide adenine dinucleotide phosphate (NADPH) and phenylalanine, substrates of the two specific oxidases. The supplementation with these substrates significantly (P < 0.05) increased the amounts of H2O2 generated from both live and dead spz, but for the two substrates, the increase was higher with dead than with live spz especially when phenylalanine was added. Addition of the antioxidant catalase significantly (P < 0.05) decreased the generation of H2O2 by live and dead spz with no significant differences (P > 0.05) between the two types of sperm before or after the antioxidant addition. This study showed the ability of live and dead ram spz to generate H2O2 in TAL medium. This ability was significantly influenced by the addition of NADPH and phenylalanine and also by the supplementation of the antioxidant catalase.
References:
Aitken R. (): Molecular mechanisms regulating human sperm function. Molecular Human Reproduction, 3, 169-173  https://doi.org/10.1093/molehr/3.3.169
 
AITKEN R. J., BUCKINGHAM D. (1992): Enhanced detection of reactive oxygen species produced by human spermatozoa with 7-dimethyl amino-naphthalin–1, 2-dicarbonic acid hydrazide. International Journal of Andrology, 15, 211-219  https://doi.org/10.1111/j.1365-2605.1992.tb01341.x
 
Aitken R. J., Clarkson J. S. (1987): Cellular basis of defective sperm function and its association with the genesis of reactive oxygen species by human spermatozoa. Reproduction, 81, 459-469  https://doi.org/10.1530/jrf.0.0810459
 
Aitken R. J., Harkiss D., Buckingham D. (1993): Relationship between iron-catalysed lipid peroxidation potential and human sperm function. Reproduction, 98, 257-265  https://doi.org/10.1530/jrf.0.0980257
 
Alomar M., Donnay I. (2006): Assessment of sperm reactive oxygen species production and oxidative stress response in different bulls. Proc. FNRS Contact Group "Competence to Development of the Mammalian Oocyte and Embryo Quality", Louvain-la-Neuve, Belgium.
 
ALVAREZ JUAN G., TOUCHSTONE JOSEPH C., BLASCO LUIS, STOREY BAYARD T. (1987): Spontaneous Lipid Peroxidation and Production of Hydrogen Peroxide and Superoxide in Human Spermatozoa Superoxide Dismutase as Major Enzyme Protectant Against Oxygen Toxicity. Journal of Andrology, 8, 338-348  https://doi.org/10.1002/j.1939-4640.1987.tb00973.x
 
Ball Barry A., Vo Anthony T., Baumber Julie (2001): Generation of reactive oxygen species by equine spermatozoa. American Journal of Veterinary Research, 62, 508-515  https://doi.org/10.2460/ajvr.2001.62.508
 
Baumber J., Sabeur K., Vo A., Ball B.A. (2003): Reactive oxygen species promote tyrosine phosphorylation and capacitation in equine spermatozoa. Theriogenology, 60, 1239-1247  https://doi.org/10.1016/S0093-691X(03)00144-4
 
Bilodeau Jean-Fran�ois, Chatterjee Suvro, Sirard Marc-Andr�, Gagnon Claude (2000): Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Molecular Reproduction and Development, 55, 282-288  https://doi.org/10.1002/(SICI)1098-2795(200003)55:3<282::AID-MRD6>3.0.CO;2-7
 
Bilodeau J.-F, Blanchette S., Gagnon C., Sirard M.-A. (2001): Thiols prevent H2O2-mediated loss of sperm motility in cryopreserved bull semen. Theriogenology, 56, 275-286  https://doi.org/10.1016/S0093-691X(01)00562-3
 
Bilodeau Jean-François, Blanchette Sophie, Cormier Nathaly, Sirard Marc-André (2002): Reactive oxygen species-mediated loss of bovine sperm motility in egg yolk Tris extender: protection by pyruvate, metal chelators and bovine liver or oviductal fluid catalase. Theriogenology, 57, 1105-1122  https://doi.org/10.1016/S0093-691X(01)00702-6
 
De Lamirande E., Gagnon C. (1992): Reactive oxygen species and human spermatozoa. I. Effects on the motility of intact spermatozoa and on sperm axonemes. Journal of Andrology, 13, 368–378.
 
de Lamirande Eve, Lamothe Geneviève (2009): Reactive oxygen-induced reactive oxygen formation during human sperm capacitation. Free Radical Biology and Medicine, 46, 502-510  https://doi.org/10.1016/j.freeradbiomed.2008.11.004
 
El-Sisy G.A., El-Nattat W.S., El-Sheshtawy R.I. (2008): Effect of superoxide dismutase and catalase on viability of cryopreserved Buffalo spermatozoa. Global Veterinaria, 2, 56–61.
 
Farrell P.B., Foote R.H., McArdle M.M., Trouern-Trend V.L., Tardif A.L. (1996): Media and dilution procedures tested to minimize handling effects on human, rabbit and bull sperm for computer-assisted sperm analysis (CASA). Journal of Andrology, 17, 293–300.
 
Fernández-Santos M. R., Domínguez-Rebolledo A. E., Esteso M. C., Garde J. J., Martínez-Pastor F. (2009): Catalase supplementation on thawed bull spermatozoa abolishes the detrimental effect of oxidative stress on motility and DNA integrity. International Journal of Andrology, 32, 353-359  https://doi.org/10.1111/j.1365-2605.2008.00871.x
 
Gavella M., Lipovac V. (1992): Nadh-Dependent Oxidoreductase (Diaphorase) Activity and Isozyme Pattern of Sperm in Infertile Men. Systems Biology in Reproductive Medicine, 28, 135-141  https://doi.org/10.3109/01485019208987691
 
Lapointe S., Sirard M.A. (1998): Catalase and oviductal fluid reverse the decreased motility of bovine sperm in culture medium containing specific amino acids. Journal of Andrology, 19, 31–36.
 
Lopes S., Jurisicova A., Sun J. G., Casper R. F. (1998): Reactive oxygen species: potential cause for DNA fragmentation in human spermatozoa. Human Reproduction, 13, 896-900  https://doi.org/10.1093/humrep/13.4.896
 
Marti E., Mara L., Marti J.I., Muiño-Blanco T., Cebrián-Pérez J.A. (2007): Seasonal variations in antioxidant enzyme activity in ram seminal plasma. Theriogenology, 67, 1446-1454  https://doi.org/10.1016/j.theriogenology.2007.03.002
 
Morales H., Tilquin P., Rees J.F., Massip A., Dessy F., Van Langendonckt A. (1999): Pyruvate prevents peroxide-induced injury of in vitro preimplantation bovine embryos. Molecular Reproduction and Development, 52, 149-157  https://doi.org/10.1002/(SICI)1098-2795(199902)52:2<149::AID-MRD5>3.0.CO;2-4
 
O'Flaherty Cristián, de Lamirande Eve, Gagnon Claude (2006): Positive role of reactive oxygen species in mammalian sperm capacitation: triggering and modulation of phosphorylation events. Free Radical Biology and Medicine, 41, 528-540  https://doi.org/10.1016/j.freeradbiomed.2006.04.027
 
Rivlin J. (2004): Role of Hydrogen Peroxide in Sperm Capacitation and Acrosome Reaction. Biology of Reproduction, 70, 518-522  https://doi.org/10.1095/biolreprod.103.020487
 
Rodriguez P.C., O’Flaherty C.M., Beconi M.T., Beorlegui N.B. (2005): Nitric oxide-induced capacitation of cryopreserved bull spermatozoa and assessment of participating regulatory pathways. Animal Reproduction Science, 85, 231-242  https://doi.org/10.1016/j.anireprosci.2004.05.018
 
Sanocka D., Kurpisz M. (2004): Reactive oxygen species and sperm cells. Reproductive Biology and Endocrinology, 2, 12–18. https://doi.org/10.1186/1477-7827-2-12
 
Shannon P., Curson B. (1982a): Kinetics of the aromatic l-amino acid oxidase from dead bovine spermatozoa and the effect of catalase on fertility of diluted bovine semen stored at 5 degrees C and ambient temperatures. Journal of Reproduction and Fertility, 64, 463–467.
 
Shannon P., Curson B. (1982b): Site of aromatic l-amino acid oxidase in dead bovine spermatozoa and determination of between-bull differences in the percentage of dead spermatozoa by oxidase activity. Journal of Reproduction and Fertility, 64, 469–473.
 
Sicherle C.C., Maia M.S., Bicudo S.D., Rodello L., Azevedo H.C. (2011): Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen supplemented with catalase or Trolox. Small Ruminant Research, 95, 144-149  https://doi.org/10.1016/j.smallrumres.2010.10.011
 
Thannickal V.J., Fanburg B.L. (2000): Reactive oxygen species in cell signaling. American Journal of Physiology – Lung Cellular and Molecular Physiology, 279, 1005–1028.
 
Upreti G.C., Jensen K., Munday R., Duganzich D.M., Vishwanath R., Smith J.F. (1998): Studies on aromatic amino acid oxidase activity in ram spermatozoa: role of pyruvate as an antioxidant. Animal Reproduction Science, 51, 275-287  https://doi.org/10.1016/S0378-4320(98)00082-7
 
download PDF

© 2020 Czech Academy of Agricultural Sciences