FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Antioxidants (Basel) / Catalase in Unexpected Places: Revisiting H2O2 Detoxificatio...
Antioxidants (Basel, Switzerland)2025; 14(6); doi: 10.3390/antiox14060718

Catalase in Unexpected Places: Revisiting H2O2 Detoxification Pathways in Stallion Spermatozoa.

Abstract: Oxidative stress plays a critical role in regulating sperm function, yet species-specific antioxidant mechanisms remain poorly understood. This study compared hydrogen peroxide (H2O2) tolerance in horse and human sperm and investigated the roles of catalase and glutathione peroxidase (GPx) in horses. A H2O2 dose-response assay (0-2000 µM) showed that horse sperm were significantly more resistant to oxidative damage, with an IC50 for progressive motility over 14-fold higher than that of human sperm (391.6 µM vs. 27.3 µM). Horse sperm also accumulated more intracellular H2O2 without loss of motility or viability. DNA damage assays (Halo and SCSA) revealed H2O2-induced fragmentation in human but not horse sperm. Enzyme inhibition experiments in horse sperm using 3-amino-1,2,4-triazole (catalase inhibitor) and (1S,3R)-RSL3 (GPx inhibitor) at 250 µM H2O2 showed that catalase inhibition severely impaired motility and increased intracellular H2O2 > 100-fold, while GPx inhibition had a milder effect (~5-fold increase). Immunocytochemistry localized catalase to the sperm head, particularly the post-acrosomal region, challenging the notion that sperm lack peroxisomes. The dependence of horse sperm on oxidative phosphorylation may drive the need for enhanced antioxidant defenses. These findings reveal species-specific oxidative stress adaptations and highlight catalase as a key antioxidant in equine sperm.
Get Full Text
Publication Date: 2025-06-12 PubMed ID: 40563349PubMed Central: PMC12190166DOI: 10.3390/antiox14060718Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses the influence of oxidative stress on sperm function, particularly comparing horse and human sperm’s tolerance to hydrogen peroxide. The study found that horse sperm are more resistant to oxidative damage and accumulate more intracellular hydrogen peroxide without losing motility or viability. This tolerance is linked to the activity of antioxidant enzymes, particularly catalase, which is identified as a key antioxidant in horse sperm.

Objective of the Study

  • The researchers aimed to understand the role of oxidative stress in regulating sperm function by comparing hydrogen peroxide tolerance between horse and human sperm. Additionally, they sought to investigate the part played by the antioxidant enzymes catalase and glutathione peroxidase (GPx) in horse sperm.

Methodology

  • They conducted a hydrogen peroxide dose-response assay to examine how varying amounts of hydrogen peroxide affects horse and human sperm.
  • To evaluate oxidative damage, they compared the sperm’s progressive motility – the ability to move in a straight line.
  • The researchers utilized DNA damage assays (Halo and SCSA) to monitor hydrogen peroxide-induced fragmentation.
  • The impact of enzyme inhibition in horse sperm was observed through the use of 3-amino-1,2,4-triazole (catalase inhibitor) and (1S,3R)-RSL3 (GPx inhibitor).
  • Finally, they used immunocytochemistry to locate catalase in the sperm, challenging the prevailing idea that sperm lacks peroxisomes (organelles that contain enzymes like catalase).

Key Findings

  • The study shows that horse sperm are more resistant to oxidative damage, and despite higher intracellular hydrogen peroxide, they retain their motility and viability.
  • This resilience can be attributed to the stronger antioxidant defences in horse sperm. Catalase, particularly, was identified as a key antioxidant enzyme.
  • When catalase in horse sperm was inhibited, it led to a sharp increase in intracellular hydrogen peroxide and a significant reduction in sperm motility. The inhibition of GPx, however, had a milder effect.
  • The study also disputes the belief that peroxisomes are absent in sperm. Catalase was localized, especially to the post-acrosomal region of the sperm head, hinting to the presence of peroxisomes.

Conclusions and Implications

  • This study reveals species-specific adaptations to oxidative stress, highlighting the relevance of catalase in equine sperm’s antioxidant defences.
  • The findings could further the understanding of the reproductive strategies in different species and enhance assisted reproductive technologies.

Cite This Article

APA
Medica AJ, Swegen A, Seifi-Jamadi A, McIntosh K, Gibb Z. (2025). Catalase in Unexpected Places: Revisiting H2O2 Detoxification Pathways in Stallion Spermatozoa. Antioxidants (Basel), 14(6). https://doi.org/10.3390/antiox14060718

Publication

Antioxidants (Basel, Switzerland)
ISSN: 2076-3921
NlmUniqueID: 101668981
Country: Switzerland
Language: English
Volume: 14
Issue: 6

Researcher Affiliations

Medica, Ashlee J
  • Discipline of Biological Sciences, School of Environmental and Life Sciences, College of Engineering Science and Environment, University of Newcastle, Callaghan, NSW 2308, Australia.
Swegen, Aleona
  • Discipline of Biological Sciences, School of Environmental and Life Sciences, College of Engineering Science and Environment, University of Newcastle, Callaghan, NSW 2308, Australia.
Seifi-Jamadi, Afshin
  • Discipline of Biological Sciences, School of Environmental and Life Sciences, College of Engineering Science and Environment, University of Newcastle, Callaghan, NSW 2308, Australia.
McIntosh, Kaitlin
  • Discipline of Biological Sciences, School of Environmental and Life Sciences, College of Engineering Science and Environment, University of Newcastle, Callaghan, NSW 2308, Australia.
Gibb, Zamira
  • Discipline of Biological Sciences, School of Environmental and Life Sciences, College of Engineering Science and Environment, University of Newcastle, Callaghan, NSW 2308, Australia.

Grant Funding

  • LP230100156 / Australian Research Council

Conflict of Interest Statement

The authors declare no conflicts of interest.

References

This article includes 45 references
  1. De Lamirande E, Leclerc P, Gagnon C. Capacitation as a regulatory event that primes sperm for the acrosome reaction and fertilization.. Mol. Hum. Reprod. 1997;3:175–194.
    doi: 10.1093/molehr/3.3.175pubmed: 9237244google scholar: lookup
  2. Aitken RJ. Molecular mechanisms regulating human sperm function.. Mol. Hum. Reprod. 1997;3:169–173.
    doi: 10.1093/molehr/3.3.169pubmed: 9237243google scholar: lookup
  3. Belen Herrero M, Chatterjee S, Lefievre L, De Lamirande E, Gagnon C. Nitric oxide interacts with the cAMP pathway to modulate capacitation of human sperm.. Free Radic. Biol. Med. 2000;29:522–536.
    doi: 10.1016/S0891-5849(00)00339-7pubmed: 11025196google scholar: lookup
  4. Aitken RJ, Harkiss D, Knox W, Paterson M, Irvine DS. A novel signal transduction cascade in capacitating human sperm characterised by a redox-regulated, cAMP-mediated induction of tyrosine phosphorylation.. J. Cell Sci. 1998;111:645–656.
    doi: 10.1242/jcs.111.5.645pubmed: 9454738google scholar: lookup
  5. Ecroyd HW, Jones RC, Aitken RJ. Endogenous redox activity in mouse sperm and its role in regulating the tyrosine phosphorylation events associated with sperm capacitation.. Biol. Reprod. 2000;69:347–354.
    doi: 10.1095/biolreprod.102.012716pubmed: 12672670google scholar: lookup
  6. Visconti PE, Bailey JL, Moore GD, Pan D, Oolds-Clarke P, Kopf GS. Capacitation of mouse sperm. I. Correlation between the capacitation state and protein tyrosine phosphorylation.. Development 1995;121:1129–1137.
    doi: 10.1242/dev.121.4.1129pubmed: 7743926google scholar: lookup
  7. Leclerc P, De Lamirand E, Gaganon C. Cyclic adenosine 3′,5′monophosphate-dependent regulation of protein tyrosine phosphorylation in relation to human sperm capacitation and motility.. Biol. Reprod. 1996;55:684–692.
    doi: 10.1095/biolreprod55.3.684pubmed: 8862788google scholar: lookup
  8. Galantino-Homer HL, Visconti PE, Kopf GS. Regulation of protein tyrosine phosphorylation during bovine sperm capacitation by a cyclic adenosine 3′5′-monophosphate-dependent pathway.. Biol. Reprod. 1997;56:707–719.
    doi: 10.1095/biolreprod56.3.707pubmed: 9047017google scholar: lookup
  9. Aitken RJ. Reactive oxygen species as mediators of sperm capacitation and pathological damage.. Mol. Reprod. Dev. 2017;84:1039–1052.
    doi: 10.1002/mrd.22871pubmed: 28749007google scholar: lookup
  10. Jones R, Mann T, Sherins R. Peroxidative breakdown of phospholipids in human sperm, spermicidal properties of fatty acid peroxides, and protective action of seminal plasma.. Fertil. Steril. 1979;31:531–537.
    doi: 10.1016/S0015-0282(16)43999-3pubmed: 446777google scholar: lookup
  11. Koppers AJ, Gargs ML, Aitken RJ. Stimulation of mitochondrial reactive oxygen species production by unesterified, unsaturated fatty acids in defective human sperm.. Free Radic. Biol. Med. 2010;48:112–119.
    doi: 10.1016/j.freeradbiomed.2009.10.033pubmed: 19837155google scholar: lookup
  12. Gibb Z, Lambourne SR, Aitken RJ. The Paradoxical Relationship Between Stallion Fertility and Oxidative Stress.. Biol. Reprod. 2014;91:77.
    doi: 10.1095/biolreprod.114.118539pubmed: 25078685google scholar: lookup
  13. Nesci S, Spinaci M, Galeati G, Nerozzi C, Pagliarana A, Algieri C, Tamanini C, Bucci D. Sperm function and mitochondrial activity: An insight on boar sperm metabolism.. Theriogenology 2020;144:82–88.
    doi: 10.1016/j.theriogenology.2020.01.004pubmed: 31927418google scholar: lookup
  14. Halliwell B, Gutteridge JM. Oxygen free radicals and iron in relation to biology and medicine: Some problems and concepts.. Arch. Biochem. Biophys. 1986;246:501–514.
    doi: 10.1016/0003-9861(86)90305-Xpubmed: 3010861google scholar: lookup
  15. Gibb Z, Lambourne SR, Curry BJ, Hall SE, Aitken RJ. Aldehyde Dehydrogenase Plays a Pivotal Role in the Maintenance of Stallion Sperm Motility.. Biol. Reprod. 2016;94:133.
    doi: 10.1095/biolreprod.116.140509pubmed: 27103446google scholar: lookup
  16. Storey BT. Mammalian sperm metabolism: Oxygen and sugar, friend and foe.. Int. J. Dev. Biol. 2008;52:427–437.
    doi: 10.1387/ijdb.072522bspubmed: 18649255google scholar: lookup
  17. Storey BT. Strategy of oxidative metabolism in bull sperm.. J. Exp. Zool. 1980;212:61–67.
    doi: 10.1002/jez.1402120109pubmed: 7411077google scholar: lookup
  18. Hereng TH, Elgstoen KB, Cederkvist FH, Eide L, Jahnsen T, Skalhegg BS, Rosendal KR. Exogenous pyruvate accelerates glycolysis and promotes capacitation in human sperm.. Hum. Reprod. 2011;26:3249–3263.
    doi: 10.1093/humrep/der317pmc: PMC3212877pubmed: 21946930google scholar: lookup
  19. Gebicka L, Krych-Madej J. The role of catalases in the prevention/promotion of oxidative stress.. J. Inorg. Biochem. 2019;197:110699.
    doi: 10.1016/j.jinorgbio.2019.110699pubmed: 31055214google scholar: lookup
  20. Alvarez JG, Storey BT. Role of glutathione peroxidase in protecting mammalian spermatozoa from loss of motility caused by spontaneous lipid peroxidation.. Gamete Res. 1989;23:77–90.
    doi: 10.1002/mrd.1120230108pubmed: 2545584google scholar: lookup
  21. Lapoint S, Sullivan R, Sirard M-A. Binding of a Bovine Oviductal Fluid Catalase to Mammalian Spermatozoa.. Biol. Reprod. 1998;58:747–753.
    doi: 10.1095/biolreprod58.3.747pubmed: 9510962google scholar: lookup
  22. Chauvin T, Xie F, Liu T, Nicora CD, Yang F, Camp DG 2nd, Smith RD, Roberts KP. A systematic analysis of a deep mouse epididymal sperm proteome.. Biol. Reprod. 2012;87:141.
    doi: 10.1095/biolreprod.112.104208pmc: PMC4435428pubmed: 23115268google scholar: lookup
  23. Amaral A, Castillo J, Estanyol JM, Ballescà JL, Ramalho-Santos J, Oliva R. Human Sperm Tail Proteome Suggests New Endogenous Metabolic Pathways.. Mol. Cell. Proteom. 2013;12:330–342.
    doi: 10.1074/mcp.M112.020552pmc: PMC3567857pubmed: 23161514google scholar: lookup
  24. Castillo J, Jodar M, Oliva R. The contribution of human sperm proteins to the development and epigenome of the preimplantation embryo.. Hum. Reprod. Update. 2018;24:535–555.
    doi: 10.1093/humupd/dmy017pubmed: 29800303google scholar: lookup
  25. Martin-Cano FE, Gaitskell-Phillips G, Oortiz-Rodriguez JM, Silva-Rodriguez A, Roman A, Rojo-Dominguez P, Alonso-Rodreiguez E, Tapia JA, Gil MC, Oortega-Ferrusola C. Proteomic profiling of horse sperm suggests changes in sperm metabolism and compromised redox regulation after cryopreservation.. J. Proteom. 2020;221:103765.
    doi: 10.1016/j.jprot.2020.103765pubmed: 32247875google scholar: lookup
  26. Greither T, Dejung M, Behre HM, Butter F, Herlyn H. The human sperm proteome-Toward a panel for male fertility testing.. Andrology 2023;11:1418–1436.
    doi: 10.1111/andr.13431pubmed: 36896575google scholar: lookup
  27. Horta Remedios M, Liang W, Gonzàlez LN, Li V, Da Ros VG, Cohen DJ, Zaremberg V. Ether lipids and a peroxisomal riddle in sperm.. Front. Cell Dev. Biol. 2023;11:1166232.
    doi: 10.3389/fcell.2023.1166232pmc: PMC10309183pubmed: 37397249google scholar: lookup
  28. Swegen A, Curry BJ, Gibb Z, Lambourne SR, Smith ND, Aitken RJ. Investigation of the horse sperm proteome by mass spectrometry.. Reproduction 2015;149:235–244.
    doi: 10.1530/REP-14-0500pubmed: 25504869google scholar: lookup
  29. Biggers JD, Whitten WK, Whittingham DG. Methods in Mammalian Embryology.. W. H. Freeman; New York, NY, USA: 1971.
  30. Medica AJ, Aitken RJ, Nicolson GL, Sheridan AR, Swegen A, De Iuliis GN, Gib Z. Glycerophospholipids protect stallion spermatozoa from oxidative damage in vitro.. Reprod. Fertil. 2021;2:199–209.
    doi: 10.1530/RAF-21-0028pmc: PMC8801026pubmed: 35118390google scholar: lookup
  31. Houston B, Curry B, Aitken RJ. Human spermatozoa possess an IL4I1 l-amino acid oxidase with a potential role in sperm function.. Reproduction 2015;149:587–596.
    doi: 10.1530/REP-14-0621pubmed: 25767141google scholar: lookup
  32. Evenson DP. Sperm Chromatin Structure Assay (SCSA(R)) for Fertility Assessment.. Curr. Protoc. 2022;2:e508.
    doi: 10.1002/cpz1.508pmc: PMC12016460pubmed: 35926128google scholar: lookup
  33. Tavares RS, Escada-Rebelo S, Correia M, Mota PC, Ramalho-Santos J. The non-genomic effects of endocrine-disrupting chemicals on mammalian sperm.. Reproduction 2016;151:R1–R13.
    doi: 10.1530/REP-15-0355pubmed: 26585413google scholar: lookup
  34. De Iuliis GN, Thomson LK, Mitchell LA, Finnie JM, Koppers AJ, Hedges A, Nixon B, Aitken RJ. DNA damage in human spermatozoa is highly correlated with the efficiency of chromatin remodeling and the formation of 8-hydroxy-2′-deoxyguanosine, a marker of oxidative stress.. Biol. Reprod. 2009;81:517–524.
    doi: 10.1095/biolreprod.109.076836pubmed: 19494251google scholar: lookup
  35. Alvarez JG, Storey BT. Differential incorporation of fatty acids into and peroxidative loss of fatty acids from phospholipids of human spermatozoa.. Mol. Reprod. Dev. 1995;42:334–346.
    doi: 10.1002/mrd.1080420311pubmed: 8579848google scholar: lookup
  36. Williams AC, Ford WCL. Functional Significance of the Pentose Phosphate Pathway and Glutathione Reductase in the Antioxidant Defenses of Human Sperm.. Biol. Reprod. 2004;71:1309–1316.
    doi: 10.1095/biolreprod.104.028407pubmed: 15189835google scholar: lookup
  37. Jeulin C, Soufir JC, Weber P, Laval-Martin D, Calvayrac R. Catalase activity in human spermatozoa and seminal plasma.. Gamete Res. 1989;24:185–196.
    doi: 10.1002/mrd.1120240206pubmed: 2793057google scholar: lookup
  38. Ball BA, Gravance CG, Medina V, Baumber J, Liu IK. Catalase activity in equine semen.. Am. J. Vet. Res. 2000;61:1026–1030.
    doi: 10.2460/ajvr.2000.61.1026pubmed: 10976731google scholar: lookup
  39. Leemans B, Gadella BM, Stout TA, De Schauwer C, Nelis H, Hoogewijs M, Van Soom A. Why doesn’t conventional IVF work in the horse? The equine oviduct as a microenvironment for capacitation/fertilization.. Reproduction 2016;152:R233–R245.
    doi: 10.1530/REP-16-0420pubmed: 27651517google scholar: lookup
  40. Felix MR, Turner RM, Dobbie T, Hinrichs K. Successful in vitro fertilization in the horse: Production of blastocysts and birth of foals after prolonged sperm incubation for capacitation.. Biol. Reprod. 2022;107:1551–1564.
    doi: 10.1093/biolre/ioac172pmc: PMC9752964pubmed: 36106756google scholar: lookup
  41. Mehdi MZ, Azar ZM, Srivastava AK. Role of receptor and nonreceptor protein tyrosine kinases in H2O2-induced PKB and ERK1/2 signaling.. Cell Biochem. Biophys. 2007;47:1–10.
    doi: 10.1385/CBB:47:1:1pubmed: 17406055google scholar: lookup
  42. O’Flaherty C, De Souza AR. Hydrogen peroxide modifies human sperm peroxiredoxins in a dose-dependent manner.. Biol. Reprod. 2011;84:238–247.
    doi: 10.1095/biolreprod.110.085712pmc: PMC4480814pubmed: 20864641google scholar: lookup
  43. Sies H. Hydrogen peroxide as a central redox signaling molecule in physiological oxidative stress: Oxidative eustress.. Redox Biol. 2017;11:613–619.
    doi: 10.1016/j.redox.2016.12.035pmc: PMC5256672pubmed: 28110218google scholar: lookup
  44. Gautam R, Priyadarshini E, Patel AK, Arora T. Assessing the impact and mechanisms of environmental pollutants (heavy metals and pesticides) on the male reproductive system: A comprehensive review.. J. Environ. Sci. Health Part C Toxicol. Carcinog. 2024;42:126–153.
    doi: 10.1080/26896583.2024.2302738pubmed: 38240636google scholar: lookup
  45. Lettieri G, Notariale R, Ambrosino A, Di Bonito A, Giarra A, Trifuoggi M, Manna C, Piscopo M. Spermatozoa Transcriptional Response and Alterations in PL Proteins Properties after Exposure of Mytilus galloprovincialis to Mercury.. Int. J. Mol. Sci. 2021;22:1618.
    doi: 10.3390/ijms22041618pmc: PMC7915165pubmed: 33562685google scholar: lookup
Subscribe to our newsletter
Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.