Validation of the CellRox Deep Red® fluorescent probe to oxidative stress assessment in equine spermatozoa
Renata Lançoni, Rubens Paes de Arruda, Maíra Bianchi Rodrigues Alves, Letícia Zoccolaro Oliveira, Gabriel De Carli dos Santos, Kleber Menegon Lemes, Shirley Andréa Florez-Rodriguez, Eneiva CarlaCarvalho Celeghini
Abstract
Considering the importance of ROS influence on sperm functionality and some limitations in sperm oxidative stress assessment methods, a field to studies of new techniques are still open. In this sense, the aim of this study is to validate the ROS detection technique through the CellRox Deep Red Reagent® probe in stallion sperm. Four stallions were used and the analyses were conducted on four replicates of semen samples from each of stallion (n = 16). The results of the polynomial regression presented a quadratic effect, high determination coefficient value (R2 = 0.88) and high significant P value (P < 0.0001). The CellRox Deep Red® fluorescent probe is able to detect reactive oxygen species in equine sperm, indicating accurately the occurrence of oxidative stress in stallion semen.
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References
Aitken RJ, Harkiss D, Buckingham D. 1993. Relationship between iron-catalysed lipid peroxidation potencial and human sperm function. J Reprod Fertil, 98:257-265.
Aitken RJ, Baker MA. 2004. Oxidative stress and male reproductive biology. Reprod Fertil Dev, 16:581- 588.
Aitken RJ, De Iuliis GN, Gibb Z, Baker MA. 2012. The simmet lecture: new horizons on an old landscape – oxidative stress, DNA damage and apoptosis in the male germ line. Reprod Domest Anim, 47(suppl. 4):7-14.
Aitken RJ, Smith TB, Lord T, Kuczera L, Koppers AJ, Naumovski N, Connaughton H, Baker MA, De Iuliis GN. 2013. On methods for the detection of reactive oxygen species generation by human spermatozoa: analysis of the cellular responses to catechol oestrogen, lipid aldehyde, menadione and arachidonic acid. Andrology, 1:192-205.
Alves MBR, De Andrade AFC, Arruda RP, Batissaco L, Florez-Rodriguez SA, Lançoni R, Oliveira BMM, Torres MA, Ravagnani GM,
Almeida TG, Vellone VS, Celeghini ECC. 2015. An efficient technique to detect sperm reactive oxygen species: the CellRox Deep Red® fluorescente probe.
Biochem Physiol, 4:1-5. Ball BA, Vo A. 2002. Detection of lipid peroxidation in equine spermatozoa based upon the lipophilic fluorescent dye C11-BODIPY581/591. J Androl, 23:259-269.
Bavister D, Lorraine M. 1983. Development of preimplantation in a defined embryos of the Golden culture medium. Biol Trace Elem Res, 28:235-247.
Brouwers JF, Silva PFN, Gadella BM. 2005. New assays for detection and localization of endogenous lipid peroxidation products in living boar sperm after BTS dilution or after freeze-thawing. Theriogenology, 63:458-469.
Gibb Z, Lambourne SR, Aitken RJ. 2014. The paradoxical relationship between stallion fertility and oxidative stress. Biol Reprod, 91:1-10.
Grinberg YY, Dibbern ME, Levasseur VA, Kraig RP. 2013. Insulin-like growth factor-1 abrogates microglial oxidative stress and TNF-a responses to spreading depression. J Neurochem, 126:662-672.
Kankofer M, Kolm G, Aurich J, Aurich C. 2005. Activity of glutathione peroxidase, superoxide dismutase and catalase and lipid peroxidation intensity in stallion sêmen during storage at 5°C. Theriogenology, 63:1354-1365.
Lenzi A, Picardo M, Gandini L, Dondero F. 1996. Lipids of the sperm plasma membrane: From polyunsaturated fatty acids considered as markers of sperm function to possible scavenger therapy. Hum Reprod Update, 2:246-256.
Li TK. 1975. The glutathione and thiol contente of mammalian spermatozoa and seminal plasma. Biol Reprod, 12:641-646.
Nichi M, Bols PEJ, Züge RM, Barnabe VH, Goovaerts IGF, Barnabe RC, Cortada CNM. 2006. Seasonal variation in sêmen quality in Bos indicus and Bos taurus bulls raised under tropical conditions. Theriogenology, 66:822-828.
Ortega-Ferrusola C, Johannisson A, Peña FJV, Tapia JA, Rodriguez-Martinez H, Dalin AM, Morrell JM. 2011. Effect of diferente extenders and seminal plasma on the susceptibility of equine spermatozoa to lipid peroxidation after single-layer centrifugation, through androcoll-E. J Equine Vet Sci, 31:411-416.
Raphael CF, Andrade AFC, Nascimento J, Arruda RP. 2008. Effects of centrifugation on membrane integrity and lipid peroxidation of equine cooled spermatozoa. Anim Reprod Sci, 107:344-345.
jWang H, Joseph J. 1999. Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader11mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee by the United States Department of Agriculture and does not imp. Free Radic Biol Med, 27:612-616.
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