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Home / Equine Research Bank / PLoS One / Inhibition of Mitochondrial Complex I Leads to Decreased Mot...
PloS one2015; 10(9); e0138777; doi: 10.1371/journal.pone.0138777

Inhibition of Mitochondrial Complex I Leads to Decreased Motility and Membrane Integrity Related to Increased Hydrogen Peroxide and Reduced ATP Production, while the Inhibition of Glycolysis Has Less Impact on Sperm Motility.

Abstract: Mitochondria have been proposed as the major source of reactive oxygen species in somatic cells and human spermatozoa. However, no data regarding the role of mitochondrial ROS production in stallion spermatozoa are available. To shed light on the role of the mitochondrial electron transport chain in the origin of oxidative stress in stallion spermatozoa, specific inhibitors of complex I (rotenone) and III (antimycin-A) were used. Ejaculates from seven Andalusian stallions were collected and incubated in BWW media at 37 °C in the presence of rotenone, antimycin-A or control vehicle. Incubation in the presence of these inhibitors reduced sperm motility and velocity (CASA analysis) (p<0.01), but the effect was more evident in the presence of rotenone (a complex I inhibitor). These inhibitors also decreased ATP content. The inhibition of complexes I and III decreased the production of reactive oxygen species (p<0.01) as assessed by flow cytometry after staining with CellRox deep red. This observation suggests that the CellRox probe mainly identifies superoxide and that superoxide production may reflect intense mitochondrial activity rather than oxidative stress. The inhibition of complex I resulted in increased hydrogen peroxide production (p<0.01). The inhibition of glycolysis resulted in reduced sperm velocities (p<0.01) without an effect on the percentage of total motile sperm. Weak and moderate (but statistically significant) positive correlations were observed between sperm motility, velocity and membrane integrity and the production of reactive oxygen species. These results indicate that stallion sperm rely heavily on oxidative phosphorylation (OXPHOS) for the production of ATP for motility but also require glycolysis to maintain high velocities. These data also indicate that increased hydrogen peroxide originating in the mitochondria is a mechanism involved in stallion sperm senescence.
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Publication Date: 2015-09-25 PubMed ID: 26407142PubMed Central: PMC4583303DOI: 10.1371/journal.pone.0138777Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 focuses on examining the role of mitochondria, particularly complex I, in reactive oxygen species (ROS) production and its correlation with motility and membrane integrity in stallion sperm. The researchers prove that the inhibition of mitochondrial complexes I and III affects sperm motility, velocity and ATP content significantly. Additionally, the study reveals the relationship between increased hydrogen peroxide production and sperm aging.

Objective and Methodology

  • The purpose of the study was to investigate the role of mitochondrial ROS (reactive oxygen species) production and its effect on motility and membrane integrity of stallion spermatozoa. The study made use of specific inhibitors of complex I (rotenone) and III (antimycin-A) of the mitochondrial electron transport chain to achieve this.
  • Ejaculates from seven Andalusian stallions were collected and incubated under varied conditions using the inhibitors at larger quantities.

Findings

  • The use of inhibitors resulted in reduced sperm motility and velocity with the effect being more evident in the use of rotenone (complex I inhibitor). The ATP content in the sperm also decreased as a result.
  • Complexes I and III were shown to decrease the production of reactive oxygen species (ROS). This implies that the superoxide production may be reflective of intense mitochondrial activity rather than oxidative stress.
  • The study found an increased production of hydrogen peroxide upon the inhibition of complex I.
  • The inhibition of glycolysis (the process of breaking down glucose) resulted in reduced sperm velocity but did not seem to affect the total percent of motile sperm.

Conclusion and Implications

  • There were correlations reported between sperm motility, velocity and membrane integrity and the production of ROS, with mitochondrial oxidative phosphorylation proofed to have a significant role in ATP production, which fuels motility.
  • Despite this, glycolysis was also necessary to maintain higher velocities, implicating both the mechanisms have roles in maintaining optimal stallion sperm functionality.
  • Increased hydrogen peroxide production was implicated in the process of sperm aging or senescence, originating in the mitochondria.

Cite This Article

APA
Plaza Davila M, Martin Muñoz P, Tapia JA, Ortega Ferrusola C, Balao da Silva C C, Peña FJ. (2015). Inhibition of Mitochondrial Complex I Leads to Decreased Motility and Membrane Integrity Related to Increased Hydrogen Peroxide and Reduced ATP Production, while the Inhibition of Glycolysis Has Less Impact on Sperm Motility. PLoS One, 10(9), e0138777. https://doi.org/10.1371/journal.pone.0138777

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 10
Issue: 9
Pages: e0138777
PII: e0138777

Researcher Affiliations

Plaza Davila, María
  • Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
Martin Muñoz, Patricia
  • Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
Tapia, Jose A
  • Department of Physiology, Faculty of Veterinary Medicine, University of Extremadura, Cáceres, Spain.
Ortega Ferrusola, Cristina
  • Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
Balao da Silva C, Carolina
  • Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.
Peña, Fernando J
  • Laboratory of Equine Reproduction and Equine Spermatology, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain.

MeSH Terms

  • Adenosine Triphosphate / biosynthesis
  • Animals
  • Electron Transport Complex I / antagonists & inhibitors
  • Glycolysis / drug effects
  • Horses
  • Hydrogen Peroxide / metabolism
  • Male
  • Membrane Potential, Mitochondrial / drug effects
  • Mitochondria / metabolism
  • Mitochondrial Membranes / metabolism
  • Oxidative Stress / drug effects
  • Reactive Oxygen Species / metabolism
  • Sperm Motility / drug effects
  • Spermatozoa / drug effects
  • Spermatozoa / metabolism

Grant Funding

  • R49 CE001002 / NCIPC CDC HHS

Conflict of Interest Statement

The authors have declared that no competing interests exist.

References

This article includes 43 references

Citations

This article has been cited 33 times.
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