Formation of Hydrogen Peroxide by Chilled Goat Spermatozoa and the Effects of Dead Spermatozoa on Motility Characteristics
Subject Areas : Camel
م.
آلومار
1
(Department of Agriculture, Atomic Energy Commission, Damascus, Syria)
Keywords: Goat, reactive oxygen species, motility, dead spermatozoa,
Abstract :
In this study, the formation of hydrogen peroxide (H2O2) by chilled goat spermatozoa was measured. Furthermore, the effects of dead spermatozoa on motility characteristics were studied. Fresh collected ejaculates from five Shami bucks were centrifuged and virtually all seminal plasma was removed. A part of the collected spermatozoa was killed by two ways: the first by repeated freezing in liquid nitrogen and thawing at 37 ˚C in water bath and the second by adding the spermatozoa to double distilled water.Two experiments were conducted after two hours of samples incubation intris-egg yolk (TEY) medium at 5 ˚C. In the first experiment, a fluorometric assay with 10-acetyl-3,7-dihydroxyphenoxazine agent as a probe for H2O2 detection was used to measure H2O2 formation. In the second experiment, the effects of adding 0 (control), 25, 50 and 75 % (V/V) of dead spermatozoa to live ones on sperm motility were assessed using computer-aided sperm analysis (CASA). Hydrogen peroxide was generated from live and dead chilled spermatozoa and the amounts of this agent increased with time. Moreover, clear significant differences (P<0.05) were observed between the generated levels from dead spermatozoa compared to live ones. The dead spermatozoa by repeated freezing-thawing treatment produced the higher H2O2 amount (P<0.05). The values of percent motile spermatozoa (MOT %), percent of progressively motile spermatozoa (PMOT %) and average path velocity (VAP) were significantly (P<0.05) reduced compared to controls when dead spermatozoa were added. The negative effect on the previous CASA parameters was increased when the percentages of dead spermatozoa were increased whatever the way of sperm death was. In conclusion, the high formation of H2O2 from dead chilled goat spermatozoa may be responsible for motility decreased. The removal of dead spermatozoa from incubation medium could help to improve the motility characteristics of chilled goat sperm.
Agarwal A., Prabakaran S. and Allamaneni S. (2006). What an andrologist/urologist should know about free radicals and why. Urology.67, 2-8.
Aitken R.J. and Buckingham D. (1992). Enhanced detection of reactive oxygen species produced by human spermatozoa with 7-dimethyl amino-naphthalin-1, 2-dicarbonic acid hydrazide. Int. J. Androl. 15, 211-219.
Aitken R.J. and Fisher H. (1994). Reactive oxygen species generation and human spermatozoa: The balance of benefit and risk. Bioessays. 16(4), 259-267.
Aitken R.J., Ryan A.L., Baker M.A. and McLaughlin E.A. (2004). Redox activity associated with the maturation and capacitation of mammalian spermatozoa. Free Radic. Biol. Med. 36, 994-1010.
Alomar M., Alzoabi M. and 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.
Alomar M. and Donnay I. (2006). Assessment of Sperm Reactive Oxygen Species Production and Oxidative Stress Response in Different Bulls. Work Shop (FNRS Contact Group), Louvain –la– Neuve, Belgium.
Alomar M., Mahieu J., Verhaeghe B., Defoin L. and Donnay I. (2006). Assessment of sperm quality parameters of six bulls showing different abilities to promote embryo development. Reprod. Fertil. Dev. 18, 395-402.
Appell R.A., Evans P.R. and Blandy J.P. (1977). The effect of temperature on the motility and viability of sperm. Br. J. Urol. 49, 751-156.
Ball B.A., Vo A.T. and Baumber J. (2001). Generation of reactive oxygen species by equine spermatozoa. American J. Vet. Res. 62, 508-515.
Bilodeau J.F., Blanchette S., Cormier N. and Sirard M.A. (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.
Brinsko S.P., Blanchard T.L., Rigby S.L., Love C.C. and Varner D.D. (2003). Effects of dead spermatozoa on motion characteristics and membrane integrity of live spermatozoa in fresh and cooled-stored equine semen. Theriogenology. 59, 735-742.
Bui A.D., Sharma R., Henkel R. and Agarwal A. (2018). Reactive oxygen species impact on sperm DNA and its role in male infertility. Andrologia. 50(8), e13012.
Chatterjee S. and Gagnon C. (2001). Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol. Reprod. Dev. 59, 451-458.
Ford W.C.L. (2004). Regulation of sperm function by reactive oxygen species. Hum. Reprod. Update. 10(5), 387-399.
Halliwell B. and Gutteridge J.M. (1984). Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem. J. 219, 1-14.
Holland M.K. and Story B.T. (1981). Oxygen metabolism of mammalian spermatozoa. Generation of hydrogen peroxide by rabbit epididymal spermatozoa. Biochem. J. 15, 273-280.
Jasko D.J., Moran D.M., Farlin M.E. and Squires E.L. (1991). Effect of seminal plasma dilution or removal on spermatozoal motion characteristics of cooled stallion semen. Theriogenology. 35, 1059-1067.
Leboeuf B., Restall B. and Salamon S. (2000). Production and storage of goat semen for artificial insemination. Anim. Reprod. Sci. 62, 113-141.
Memon A.A., Wahid H., Rosnina Y., Goh Y.M., Ebrahimi M. and Nadia F.M. (2013). Effect of ascorbic acid concentrations, methods of cooling and freezing on boer goat semen cryopreservation. Reprod. Domest. Anim. 48, 325-330.
Memon M.A., Bretzlaff K.N. and Ott R.S. (1985). Effect of washing on motility and acrosome morphology of frozen-thawed goat spermatozoa. American J. Vet. Res. 46, 473-475.
Mortimer S.T. (2000). CASA-practical aspects. J. Androl. 21, 515-524.
Nair S.J., Brar A.S., Ahuja C.S., Sangha S.P. and Chaudhary K.C. (2006). Comparative study on lipid peroxidation, activities of antioxidant enzymes and viability of cattle and buffalo bull spermatozoa during storage at refrigeration temperature. Anim. Reprod. Sci. 96, 21-29.
O'Flaherty C., De Lamirande E. and Gagnon C. (2006). Positive role of reactive oxygen species in mammalian sperm capacitation: Triggering and modulation of phosphorylation events. Free Radic. Biol. Med. 41, 528-540.
Paulenz H., Soderquist L., Perez-Pe´ R. and Breg K.A. (2002). Effect of different extenders and storage temperatures on sperm viability of liquid ram semen. Theriogenology. 57, 823-836.
Pruitt J.A., Arns M.J. and Pool K.C. (1993). Seminal plasma influences recovery of equine spermatozoa following in vitro culture (37 ˚C) and cold-storage (5 ˚C). Theriogenology. 39, 291-296.
Purdy P.H. (2006). A review on goat sperm cryopreservation. Small Rumin. Res. 63, 215-225.
Rodriguez P.C., O'Flaherty C.M., Beconi M.T. and Beorlegui N.B. (2005). Nitric oxide-induced capacitation of cryopreserved bull spermatozoa and assessment of participating regulatory pathways. Anim. Reprod. Sci. 85, 231-242.
Salamon S. and Maxwell W.M.C. (1995). Frozen storage of ram semen: II. Causes of low fertility after cervical insemination and methods of improvement. Anim. Reprod. Sci. 38, 1-36.
Shannon P. and 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 ˚C and ambient temperatures. J. Reprod. Fertil. 64, 463-467.
Shannon P. and 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. J. Reprod. Fertil. 64, 469-473.
Sanocka D. and Kurpisz M. (2004). Reactive oxygen species and sperm cells. Reprod. Biol. Endocrinol. 2, 12-18.
Slavador I., Yaniz J., Viudes-de-Castro M.P., Gomez E.A. and Silvestre M.A. (2006). Effect of solid storage on caprine semen conservation at 5 ˚C. Theriogenology. 66, 974-981.
Todd P., Arns M.J., Chenoweth P. and Schultz B. (2001). Influence of seminal plasma and processing on cold stored stallion spermatozoa. Anim. Reprod. Sci. 68, 335-336.
Trokey D.E. and Merilan C.P. (1982). Effect of added cold shocked cells upon the viability of pony stallion spermatozoa. Theriogenology. 18, 723-725.
Upreti G.C., Jensen K., Munday R., Duganzich D.M., Vishwanath R. and Smith J.F. (1998). Studies on aromatic amino acid oxidase activity in ram spermatozoa: role of pyruvate as an antioxidant. Anim. Reprod. Sci. 51, 275-287.
Vishwanath R. and Shannon P. (1997). Do sperm cells age? A review of the physiological changes in sperm during storage at ambient temperature. Reprod. Fertil. Dev. 9, 321-331.
Wagner H., Cheng J.W. and Ko E.Y. (2018). Role of reactive oxygen species in male infertility: An updated review of literature. Arab. J. Urol. 16, 35-43.