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Home / Equine Research Bank / Animals (Basel) / Binding of Equine Seminal Lactoferrin/Superoxide Dismutase (...
Animals : an open access journal from MDPI2022; 13(1); 52; doi: 10.3390/ani13010052

Binding of Equine Seminal Lactoferrin/Superoxide Dismutase (SOD-3) Complex Is Biased towards Dead Spermatozoa.

Abstract: Sperm-neutrophil binding is an important facet of breeding and significantly impacts fertility. While a specific seminal plasma protein has been found to reduce this binding and improve fertility (CRISP-3), additional molecule(s) appear to promote binding between defective sperm and neutrophils. Recent work has suggested one of these proteins is lactoferrin (LF), an 80 kDa iron-binding protein found throughout the body, but the purity of the protein was not confirmed. It is unknown if LF binds to sperm selectively based on viability, and if receptors for LF are located on equine sperm. To evaluate this, we attempted to purify equine seminal LF from five stallions (n = 5), biotinylate LF, and evaluate potential binding site(s) on spermatozoa. LF was consistently associated with superoxide dismutase (SOD-3), and all attempts to separate the two proteins were unsuccessful. Flow cytometric and microscopic analyses were used to compare LF/SOD-3 binding to viable and nonviable spermatozoa. Additionally, various methods of biotinylation were assessed to optimize this methodology. Biotinylation of seminal plasma protein was an effective and efficient method to study seminal plasma protein properties, and the binding site for LF/SOD-3 was found to be broadly localized to the entire sperm cell surface as well as selective towards nonviable/defective sperm. Although we were not able to determine if the binding to equine spermatozoa was through LF or SOD-3, we can conclude that equine seminal LF is tightly bound to SOD-3 and this protein complex binds selectively to nonviable spermatozoa, possibly to mark them for elimination by neutrophil phagocytosis.
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Publication Date: 2022-12-23 PubMed ID: 36611662PubMed Central: PMC9817809DOI: 10.3390/ani13010052Google Scholar: Lookup
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  • 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.

This study investigates the role of lactoferrin and superoxide dismutase (SOD-3) proteins found in equine seminal plasma and their preference in binding to either viable or nonviable sperm cells. The study found that these protein complexes selectively bind to nonviable spermatozoa, most likely flagging them for elimination by neutrophil cells, the body’s primary immune response.

Introduction and Objectives

  • The research sheds light on the role of proteins like lactoferrin (an iron-binding protein) and superoxide dismutase (SOD-3) in equine fertility, specifically, their interaction with sperm-neutrophil binding, which significantly impacts fertility.
  • The main objective was to understand if lactoferrin binds selectively to viable or nonviable sperm, and the location of lactoferrin receptors on equine sperm.

Methodology

  • The researchers purified equine seminal lactoferrin from samples taken from five stallions and biotinylated it to evaluate potential binding sites on spermatozoa.
  • Throughout the study, lactoferrin was consistently found to be associated with an enzyme called superoxide dismutase (SOD-3). The researchers carried out various biotinylation methods in order to optimize the methodology to study these seminal plasma protein properties.

Findings

  • Flow cytometric and microscopic analyses conducted to compare the binding of lactoferrin and SOD-3 to viable and nonviable spermatozoa showed that these proteins bind selectively to nonviable or defective sperm.
  • The binding site for the lactoferrin/SOD-3 complex was found across the entire sperm cell surface. Nevertheless, it remained unclear whether the binding to equine spermatozoa was through lactoferrin or SOD-3.

Conclusions

  • Despite the uncertainty about which of the two proteins binds to the sperm, the study established that lactoferrin in equine seminal fluid tightly binds to SOD-3. This protein complex showed an affinity to bind selectively towards nonviable spermatozoa, likely marking them for elimination via the immune response mechanism of neutrophil phagocytosis.

Cite This Article

APA
Alghamdi AS, Fedorka CE, Scoggin KE, Esteller-Vico A, Beatty K, Davolli G, Ball BA, Troedsson MHT. (2022). Binding of Equine Seminal Lactoferrin/Superoxide Dismutase (SOD-3) Complex Is Biased towards Dead Spermatozoa. Animals (Basel), 13(1), 52. https://doi.org/10.3390/ani13010052

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 13
Issue: 1
PII: 52

Researcher Affiliations

Alghamdi, Abdorrahman S
  • Departments of Agriculture and Natural Resources, University of Minnesota Crookston, Crookston, MN 56716, USA.
Fedorka, Carleigh E
  • Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Scoggin, Kirsten E
  • Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Esteller-Vico, Alejandro
  • Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Beatty, Kaylin
  • Departments of Agriculture and Natural Resources, University of Minnesota Crookston, Crookston, MN 56716, USA.
Davolli, Gabriel
  • Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Ball, Barry A
  • Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.
Troedsson, Mats H T
  • Gluck Equine Research Center, University of Kentucky, Lexington, KY 40546, USA.

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

This article has been cited 1 times.
  1. Zheng M, Liu Y, Zhang G, Yang Z, Xu W, Chen Q. The Applications and Mechanisms of Superoxide Dismutase in Medicine, Food, and Cosmetics. Antioxidants (Basel) 2023 Aug 27;12(9).
    doi: 10.3390/antiox12091675pubmed: 37759978google scholar: lookup
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