Impact of Reactive Oxygen Species on Spermatozoa: ABalancing Act between Beneficial and Detrimental Effects

Document Type : Review Article


Male Reproduction Laboratory, PRSM Division,Central Institute for Research on Goats, Makhdoom,Farah, Mathura, 281122, India


Reactive oxygen species (ROS)plays an important role in sperm motility. The physiological generation at low concentration induces beneficial effects on sperm functions and plays a significant role in sperm metabolism. Meanwhile, the excessive generation of reactive oxygen species can overwhelm protective mechanism and triggers changes in lipid and protein layers of sperm plasma membrane, which induces lipid damage, protein damage, DNA damage, motility impairment and alteration in capacitation and acrosome reaction. Reactive oxygen species (ROS) can be measured by variable biomarkers as melondialdehyde (MDA) concentration, individual free radicals, etc. The quantification of free radicalsin livestock semen is also briefly reported.


AgarwalA., Gupta S. and Sharma S.K. (2005). Role of oxidative stress in female reproduction. Reprod. Biol. Endocrinol. 3, 28-35.
AgarwalA., Ramadan A. and Mohamed A.B. (2003). Role of reactive oxygen species in the pathophysiology of human reproduction. Fertil. Steril. 79, 829-843.
AitkenR.J. (1995). Free radicals, lipid peroxidation and sperm function. Reprod. Fertil. Devel. 7, 659-668.
AitkenR.J., Buckingham D. and Harkiss D. (1993). Use of a xanthine oxidase free radical generating system to investigate the cytotoxic effects of reactive oxygen species on human spermatozoa. J. Reprod. Fertil. 97, 441-450.
AlvarezJ.G. and Storey B.T. (1992). Evidence for increased lipid peroxidative damage and loss of superoxide dismutase activity as a mode of sub lethal cryodamage to human sperm during cryopreservation. J. Androl. 13, 232-241.
BabiorB.M., Kipnes R.S. and Curnutte J.T. (1973). The production by leukocytes of superoxide, apotencial bactericidialagent.J. Clin. Invest. 52, 741-744.
BansalA.K. and Bilaspuri G.S. (2008). Oxidative stress alters membrane sulfhdryl status and phopholipid contents of crssbreed cattle bull spermatozoa. Anim. Reprod. Sci. 104, 398-404.
BansalA.K. and Bilaspuri G.S. (2009). Antioxidant effect of vitamin E on motility, viability and lipid peroxidation of cattle spermatozoa under oxidative stress. Anim. Sci. Pap. Rep. 27, 5-14.
BaumberJ., Ball B.A., Gravance C.G., Medina V. and Davies-Morel M.C.G. (2000). The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential and membrane lipid peroxidation. J. Androl. 21,895-902.
BaumberJ., Ball B.A., Linfor J.J. and Meyers S.A. (2003). Reactive oxygen species and cryopreservation promote DNA fragmentation in equine spermatozoa. J. Androl. 24, 621-628.
BilodeauJ.F., Chatterjee S., Sirard M.A. and Gagnon C. (2000). Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Mol. Reprod. Dev. 55, 282-288.
BlakeD.R., Allen R.E. and Lunec V. (1987). Free radicals in biological systems a review orientated to inflammatory processes. Br. Med. Bull. 43, 371-385.
BucakM.N., Atessahin A., Varish O., Yuce A., Tekin N. and Akcay A. (2007). The influence of trehalose, taurine, custemine and hyaluronan on ram semen. Theriogeny. 67, 1060-1067.
BucakM.N., Sariozkan S., Tuncera P.B., Sakin F., Atessahin A., Ahindkulaksize R. and Mesut Cevik M. (2010). The effect of antioxidants on post-thawed Angora goat (Capra hircus ancryrensis) sperm parameters, lipid peroxidation and antioxidant activities. Small Rum. Res. 89, 24-30.
BucakM.N., Sariozkan S., Tuncera P.B., Ulutas P.A. and Akcadag H.I. (2009). Effect of antioxidants on microscopic semen parameters, lipid peroxidation and antioxidant activities in Angora goat semen following cryopreservation.Small Rum. Res.81, 90-95.
ChatterjeeS. and Gagnon C. (2001). Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol. Reprod. Dev. 59, 451-458.
De LamirandeE. and Gagnon C. (1992). Reactive oxygen species and human spermatozoa. 1. Effects on the motility of intact spermatozoa and on sperm axonemes. J. Androl. 13, 368-378.
FerrandiB., Cremonesi F., Consiglio A.L., Carnevali A. and Porcelli F. (1992). Cytophotometric assay of cytochrome oxidase, lactate dehydrogenase and glucose-6-phosphate-dehydrogenase activities in human peroxidized spermatozoa. Act. Histoch. 93, 363-370.
FordW.C. (2004). Regulation of sperm functions by reactive oxygen species. Hum. Reprod. Update. 10, 387-399.
FormanH.J. and Boveris A. (1984). Superoxide radical and hydrogen peroxide in mitochondria. Pp. 65-90 in Free Radicals in Biology. W.A. Pryor, Ed. Academic Press, New York.
GriveauJ.F. and Le Lannou D. (1997). Reactive oxygen species and human spermatozoa: physiology and pathology. Int. J. Androl. 20, 61-69.
GriveauJ.F., Renard P., Le Lannou D. (1997).An in vitro promoting role for hydrogen peroxide in human sperm capacitation. Int. J. Androl. 56, 24-31.
Griveau J.F., Dumont E., Renard P. and Le Lannou D. (1995). Reactive oxygen species, lipid peroxidation and enzymatic defence system in human spermatozoa. J. Reprod. Fertil. 103, 17-26.
HerreraE. and Barbas C. (2001). Vitamin E: action, metabolism and perspectives. J. Physiol. Biochem. 57, 43-56.
KimJ.G. and Parthasarathy S. (1998). Oxidation and the spermatozoa. Semin. Reprod. Endocrinol. 16, 235-239.
KumarS. and Das G.K. (2005). Frozen sperm quality with reference to reactive oxygen species: a review. Indian J. Anim. Sci. 75, 874-84.
KumarS., Garg A. and Paudel K.P. (2002). Effect of reactive oxygen species (ROS) on spermatozoa: an overview. Livest. Int. 6, 8-18.
KumaresanA., Kadirvel G., Bajarbaruah K.M., Bardolar R.K., Das A., Kumar S. and Naskar S. (2009). Preservation of boar semen at 18 oCinduces lipid peroxidation and apoptosis like changes in spermatozoa. Anim. Reprod. Sci. 110, 162-171.
Lenzi A., Picardo M., Gandini1 L. and 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.
MakkerK., Agarwal A. and Sharma R. (2009). Oxidative stressand male infertility. Indian J. Med. Res. 129, 357-367.
MazzilliF., Rossi T., Marchesini M., Ronconi C. and Dondero F. (1994). Superoxide anion in human semen related to seminal parameters and clinical aspects. Fertil. Steril. 62, 862-868.
Michael A.J., Alexopoulos C., Pontiki E.A., Hadjipavlou Litinan D., Saratsis P., Ververidis H.N. and Boscos C.M. (2007). Effect of antioxidant supplementation in semen extenders on semen quality and reactive oxygen species of frozen-thawed canine spermatozoa. Theriogenolgy. 68, 204-212.
MichaelA.J., Alexopoulos C., Pontiki E.A., Hadjipavlou Litina D., Saratsis P., Ververidis H.N. and Boscos C.M. (2009). Eff-
ect of antioxidant supplementation in semen extenders on semen quality and reactive oxygen species of chilled canine spermatozoa. Anim. Reprod. Sci. 112, 119-135.
NashT. (1953). The colorimetric estimation of formaldehyde by means of the Hantzch reaction. Biochem. J. 55, 416-421.
NogushiN. and Niki E. (1999). Chemistry of active oxygen species and antioxidants. Pp. 3-20 in Antioxidant Status, Diet, Nutrition and Health. A.M. Papas, Ed. CRC Press. Washington, DC., USA.
PontikiE. and Hadjipavlou Litina D. (2006). Antioxidant and anti-inflammatory activity of anyl-acetic and hydroxaemicacids as novel lipoxygenase inhibitors. Med. Chem. 2(3), 251-264.
PriyadharshniR., Saraswat S., Sushma Y., Ramachandran N., Jindal S.K., Kharche S.D. and Goel A.K. (2012). Influence of exogenous antioxidants on motility %, membrane integrity and lipid peroxidation of frozen thawed Sirohi buck spermatozoa. Indian J. Anim. Sci. 21, 51-57.
SaraswatS., Jindal S.K., Priyadharsini R., Ramachandran N., Yadav S., Rout P.K., Kharche S.D. and Goel A.K. (2012a). The effect of antioxidants supplementation to cryopreservation protocol on seminal attributes and sperm membrane characteristics in Sirohi goat. J. Physiol. Pharmacol. Adv. 2, 49-58.
SaraswatS., Jindal S.K., Ramachandranm N., Yadav S. and Priyardshini R. (2012b). Standardization of antioxidants fortification in frozen buck semen. Indian J. Small Rumin. 18, 47-51.
SaraswatS., Priyadharsini R., Jindal S.K., Yadav S., Ramachandran N., Kharche S.D. and Goel A.K. (2012c). Effect of antioxidants supplementation at refrigeration temperature on sperm motion characteristics and membrane integrity of Sirohi buck semen. J. Physiol. Pharmacol. Adv. 2, 77-86.
SchraufstatterI., Hyslop P., Hirshaw D., Spragg R., Sklav L. and Cochrane C. (1986). Hydrogen peroxide induced injury of cells and its prevention by inhibitors of poly (ADO-ribose) polymerase. Proc. Natl. Acad. Sci. 83, 4908-4912.
ShannonP. and Curson B. (1972). Toxic effect and action of dead sperm on diluted bovine semen. J. Dairy Sci. 55, 617-620.
SikkaS.C., Rajasekaran M. and Hellstrom W.J. (1995). Role of oxidative stress and antioxidants in male infertility. J. Androl. 16, 464-481.
SikkaS.C. (1996). Oxidative stress and role of antioxidants in normal and abnormal sperm function. Front. Biosci. 1, 78-86.
SilvaP.F.N. (2006). Physiology of peroxidation process in mammalian sperm. Ph D. Thesis. Utrecht Univ., Ridderprint, Ridderkerk.
SlaterT.F. (1984). Free radical mechanisms in tissue injury. Biochem. J. 222, 1-15.
StoreyB.T. (1997). Biochemistry of the induction and prevention of lipoperoxidative damage in human spermatozoa. Mol. Hum. Reprod. 3, 203-213.
SuraiP.F., Fujhara N., Speake B.K., Brillard J.P., Wishart G.J. and Sparks N.H.C. (2001). Polyunsaturated fatty acids, lipid peroxidation and antioxidant protection in avian semen: review. Asian-australas J. Anim. Sci. 14, 1024-1050.
TraberM.G. and Atkinson J. (2007). Vitamin E, antioxidant and nothing more. Free Radical Biol. Med. 43, 4-15. 
TselkasK., Saratsis Ph., Karagianidis A. and Samouilidis S. (2000). Extracellular presence of reactive oxygen species (ROS) in fresh and frozen-thawed canine semen and their effects on some semen parameters. Dtsch. Tierarztl. Wschr. 107, 69-72.
UpretiG.C., Jensen K., Munday R., Viswanath R. and Smith J.F. (1994a). Studies on ram spermatozoa aromatic amino acid oxidase. Proc. Aust. Soc. Reprod. Biol. 26, 115-121.
UpretiG.C., Jensen K., Munday R., Viswanath R. and Smith J.F. (1994b). Interaction of aromatic amino acid oxidase activity with components of ram diluent (RSD-1). Pp. 7-28 in Proc. 7thInt. Symp.onSpermatology, Abstr.
Wang X. and Quinn P.J. (1999). Phase separations of α-tocopherol in aqueous dispersions of distearoylphosphatidylethanolamine. Chem. Phys. Lipid. 114, 1-9.
ZhangW., Yi K., Chen C., Houa X. and Zhou X. (2012). Application of antioxidants and centrifugation for cryopreservation of boar spermatozoa. Anim. Reprod. Sci. 132, 123-128.
  • Receive Date: 20 January 2013
  • Revise Date: 05 March 2013
  • Accept Date: 16 March 2013
  • First Publish Date: 01 March 2014