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Home / Equine Research Bank / Mol Reprod Dev / Lipid peroxide formation in relation to membrane stability o...
Molecular reproduction and development2005; 72(2); 230-238; doi: 10.1002/mrd.20322

Lipid peroxide formation in relation to membrane stability of fresh and frozen thawed stallion spermatozoa.

Abstract: In this study we used a new method to detect reactive oxygen species (ROS) induced damage at the level of the sperm plasma membrane in fresh and frozen-thawed stallion sperm. Lipid peroxidation (LPO) in sperm cells was assessed by a fluorescent assay involving the labeling of stallion sperm with the LPO reporter probe C11-BODIPY(581/591). The peroxidation dependent spectral emission shift of this membrane probe could be localized using inverted spectral confocal microscopy and quantified on living and deteriorated sperm cells using flow cytometry. Mass spectrometric analysis of the main endogenous lipid class, phosphatidylcholine (PC), was carried out to determine the formation of hydroxy- and hydroperoxyphosphatidylcholine in fresh sperm cells. Peroxidation as reported by the fluorescent probe corresponded with the presence of hydroxy- and hydroperoxyphosphatidylcholine in the sperm membranes, which are early stage products of LPO. This allowed us to correlate endogenous LPO with localization of this process in the living sperm cells. In absence of peroxidation inducers, only relatively little peroxidation was noted in fresh sperm cells whereas some mid-piece specific probe oxidation was noted for frozen-thawed sperm cells. After induction of peroxidation in fresh and frozen-thawed sperm cells with the 0.1 mM of lipid soluble ROS tert-butylhydrogen peroxide (t-BUT) intense probe oxidation was produced in the mid-piece, whereas the probe remained intact in the sperm head, demonstrating antioxidant activity in the head of fresh sperm cells. At higher levels of t-BUT, probe peroxidation was also noted for the sperm head followed by a loss of membranes there. Frozen-thawed sperm were more vulnerable to t-BUT than fresh sperm. The potential importance of the new assays for sperm assessments is discussed.
(c) 2005 Wiley-Liss, Inc.
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Publication Date: 2005-06-11 PubMed ID: 15948163DOI: 10.1002/mrd.20322Google Scholar: Lookup
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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 researchers developed a method to detect reactive oxygen species (ROS) damage in sperm cells, comparing fresh and frozen-thawed samples. Their findings suggested that frozen-thawed sperm are more vulnerable to ROS damage than fresh ones.

Method for Detecting ROS Damage

  • The study presented a novel technique for identifying damage in sperm cells caused by reactive oxygen species (ROS), a type of unstable molecule that can cause damage at the cellular level.
  • The researchers tested both fresh and frozen-thawed stallion sperm for ROS-induced lipid peroxidation (LPO), a process that can greatly impact sperm cells’ functionality.
  • They used a fluorescent assay involving a lipid peroxidation reporter probe known as C11-BODIPY(581/591). This probe’s spectral emission shifts when lipid peroxidation occurs, making it ideal for identifying and quantifying ROS-related damage in living cells.

Membrane Damage Analysis

  • Using confocal microscopy, the researchers tapped into the probe’s spectral shift capability to localize the damage on both live and damaged sperm cells.
  • Flow cytometry was used for quantification, helping the team note the intensity and ratio of induced damage.
  • Mass spectrometric analysis was conducted on phosphatidylcholine (PC), the main endogenous lipid class in the sperm cells, to track the formation of related compounds during LPO.

Findings on Peroxidation

  • One observation was that lipid peroxidation as reported by the fluorescent probe matched the presence of two compounds—hydroxy- and hydroperoxyphosphatidylcholine—in the sperm membranes. These compounds are early-stage products of LPO.
  • In fresh sperm cells devoid of peroxidation inducers, there was minimal peroxidation. In contrast, frozen-thawed sperm cells showed some oxidation, specifically in the mid-piece.
  • When researchers induced peroxidation using lipid-soluble ROS tert-butylhydrogen peroxide (t-BUT), a significant level of probe oxidation occurred in the mid-piece of fresh and frozen-thawed sperm cells. However, the sperm head remained largely unaffected initially due to its antioxidant activity.

Implications and Conclusions

  • At higher volumes of t-BUT, peroxidation was also observed in the sperm head, causing loss of membranes. Frozen-thawed sperm cells were noted to be more prone to t-BUT than fresh ones.
  • Given these results, the researchers suggest their assays could have important implications for sperm assessments, particularly in relation to studies on the robustness of fresh vs. frozen-thawed sperm and the impacts of ROS.

Cite This Article

APA
Neild DM, Brouwers JF, Colenbrander B, Agüero A, Gadella BM. (2005). Lipid peroxide formation in relation to membrane stability of fresh and frozen thawed stallion spermatozoa. Mol Reprod Dev, 72(2), 230-238. https://doi.org/10.1002/mrd.20322

Publication

Molecular reproduction and development
ISSN: 1040-452X
NlmUniqueID: 8903333
Country: United States
Language: English
Volume: 72
Issue: 2
Pages: 230-238

Researcher Affiliations

Neild, D M
  • Department of Medicine, School of Veterinary Sciences, University of Buenos Aires, Argentina.
Brouwers, J F H M
    Colenbrander, B
      Agüero, A
        Gadella, B M

          MeSH Terms

          • Animals
          • Cell Membrane / metabolism
          • Freezing
          • Horses
          • Lipid Peroxides / biosynthesis
          • Lipid Peroxides / metabolism
          • Male
          • Semen Preservation
          • Spermatozoa / cytology
          • Spermatozoa / metabolism
          • tert-Butylhydroperoxide / metabolism

          Citations

          This article has been cited 16 times.
          1. Dos Reis WVA, Pereira RR, Vieira M, da Cunha CCT, Acácio BR, Macedo GG, da Costa-E-Silva EV, Sampaio BFB. Impact of quercetin, carnosine, and ozone in the cryopreservation on Nellore (Bos indicus) semen. Anim Reprod 2023;20(1):e20220048.
            doi: 10.1590/1984-3143-AR2022-0048pubmed: 37034117google scholar: lookup
          2. Swelum AA, Ba-Awadh HA, Olarinre IO, Saadeldin IM, Alowaimer AN. Effects of adding mixed chicken and quail egg yolks to the cryodiluent on the quality of ram semen before and after cryopreservation. Front Vet Sci 2022;9:1013533.
            doi: 10.3389/fvets.2022.1013533pubmed: 36311647google scholar: lookup
          3. Martínez-Barbitta M, Rivera Salinas C. Evaluation of Chilled Dog Semen Extended With Sperm Activator. Front Vet Sci 2021;8:764750.
            doi: 10.3389/fvets.2021.764750pubmed: 35224070google scholar: lookup
          4. Mańkowska A, Brym P, Paukszto Ł, Jastrzębski JP, Fraser L. Gene Polymorphisms in Boar Spermatozoa and Their Associations with Post-Thaw Semen Quality. Int J Mol Sci 2020 Mar 10;21(5).
            doi: 10.3390/ijms21051902pubmed: 32164368google scholar: lookup
          5. Zhang M, Oldenhof H, Sydykov B, Bigalk J, Sieme H, Wolkers WF. Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity. Sci Rep 2017 Jul 24;7(1):6198.
            doi: 10.1038/s41598-017-06542-zpubmed: 28740099google scholar: lookup
          6. Jeannerat E, Janett F, Sieme H, Wedekind C, Burger D. Quality of seminal fluids varies with type of stimulus at ejaculation. Sci Rep 2017 Mar 13;7:44339.
            doi: 10.1038/srep44339pubmed: 28287188google scholar: lookup
          7. Sharma L, Pandey V, Nigam R, Saxena A, Swain DK, Yadav B. Association of oxidative status and semen characteristics with seminal plasma proteins of buffalo semen. Iran J Vet Res 2016 Fall;17(4):226-230.
            pubmed: 28224004
          8. Viskupicova J, Blaskovic D, Galiniak S, Soszyński M, Bartosz G, Horakova L, Sadowska-Bartosz I. Effect of high glucose concentrations on human erythrocytes in vitro. Redox Biol 2015 Aug;5:381-387.
            doi: 10.1016/j.redox.2015.06.011pubmed: 26141922google scholar: lookup
          9. Best BP. Cryoprotectant Toxicity: Facts, Issues, and Questions. Rejuvenation Res 2015 Oct;18(5):422-36.
            doi: 10.1089/rej.2014.1656pubmed: 25826677google scholar: lookup
          10. Martorana K, Klooster K, Meyers S. Suprazero cooling rate, rather than freezing rate, determines post thaw quality of rhesus macaque sperm. Theriogenology 2014 Feb;81(3):381-8.
            doi: 10.1016/j.theriogenology.2013.10.008pubmed: 24239181google scholar: lookup
          11. McCarthy MJ, Baumber J, Kass PH, Meyers SA. Osmotic stress induces oxidative cell damage to rhesus macaque spermatozoa. Biol Reprod 2010 Mar;82(3):644-51.
            doi: 10.1095/biolreprod.109.080507pubmed: 19846599google scholar: lookup
          12. Boni R, Ruggiero R, De Luca F, Serritella ML, Di Palma T, Cecchini Gualandi S. Repeatability of Selected Parameters Related to Stallion Sperm Quality and Cryotolerance. Animals (Basel) 2025 Sep 26;15(19).
            doi: 10.3390/ani15192805pubmed: 41096400google scholar: lookup
          13. Boni R, Ruggiero R, Di Palma T, Ferrara MA, Preziosi G, Cecchini Gualandi S. Stallion Sperm Freezing with Different Extenders: Role of Antioxidant Activity and Nitric Oxide Production. Animals (Basel) 2024 Aug 25;14(17).
            doi: 10.3390/ani14172465pubmed: 39272250google scholar: lookup
          14. Zhang L, Wang X, Jiang C, Sohail T, Sun Y, Sun X, Wang J, Li Y. Effects of Different Diluents and Freezing Methods on Cryopreservation of Hu Ram Semen. Vet Sci 2024 Jun 3;11(6).
            doi: 10.3390/vetsci11060251pubmed: 38921998google scholar: lookup
          15. Neuman NM, Orzołek A, Steiner-Bogdaszewska Ż, Dziekońska A. Changes in the Morphology and Antioxidant Status of European Red Deer Sperm Stored in the Epididymides and in a Liquid State. Animals (Basel) 2024 May 31;14(11).
            doi: 10.3390/ani14111653pubmed: 38891701google scholar: lookup
          16. Catalán J, Yánez-Ortiz I, Torres-Garrido M, Ribas-Maynou J, Llavanera M, Barranco I, Yeste M, Miró J. Impact of Seminal Plasma Antioxidants on DNA Fragmentation and Lipid Peroxidation of Frozen-Thawed Horse Sperm. Antioxidants (Basel) 2024 Mar 6;13(3).
            doi: 10.3390/antiox13030322pubmed: 38539855google scholar: lookup
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