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Equine veterinary journal2021; 54(2); 415-426; doi: 10.1111/evj.13457

Seminal plasma, and not sperm, induces time and concentration-dependent neutrophil extracellular trap release in donkeys.

Abstract: In several mammalian species, acute endometritis driven by the recruitment of polymorphonuclear cells (PMN) occurs in response to semen. These PMNs release DNA to form neutrophil extracellular traps (NETs) in cattle, horse and human, leading to sperm entrapment. While there is no evidence of this phenomenon occurring in donkeys, artificial insemination (AI) with frozen-thawed semen, which results in very poor pregnancy rates, leads to a large PMN recruitment to the uterus. Objective: To investigate whether donkey semen can trigger NET release (NETosis) and if excessive NETosis occurs in response to frozen-thawed semen. Methods: In vitro experiments. Methods: Jenny PMNs were exposed to jackass fresh or frozen-thawed semen, isolated sperm or seminal plasma (SP), over the course of three experiments. NET formation in response to different treatments was assessed through manual quantification of stained slides. A one-way analysis of variance (ANOVA), followed by a post hoc Sidak test, was carried out to determine statistical significance. Results: NET release occurred in a semen concentration- and incubation-time-dependent manner. Surprisingly, frozen-thawed donkey sperm did not increase NETosis rate in comparison with the control (23 ± 2.5% vs. 31 ± 3.7%; P > .05), whereas fresh semen exposure did (78 ± 5.7% vs. 26 ± 3.2%, P < .01). NETosis increased in the presence of SP, regardless of the presence or absence of sperm, in comparison with the control in both fresh (84 ± 5.2% and 77 ± 5.0% vs. 12 ± 2.7%, respectively; P < .01) and frozen (95 ± 2.2% and 94 ± 2.9% vs. 14 ± 3.8%, respectively; P < .01) samples. Moreover, exposure of PMN to viable and motile sperm, in the absence of SP, did not increase NETosis rates (P > .05). Conclusions: Donkey SP, and not sperm-intrinsic factors, is able to trigger NETosis in both time- and semen concentration-dependent manner. The physiological relevance of such response against semen in the donkey remains to be elucidated.
© 2021 EVJ Ltd.
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Publication Date: 2021-06-01 PubMed ID: 33908643DOI: 10.1111/evj.13457Google 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 research aims to understand whether neutrophil extracellular trap (NET) release is triggered by donkey semen and to ascertain whether or not excessive NET release occurs in response to frozen-thawed semen.

Objective

The main objective of this study was to investigate whether or not donkey semen could catalyze the release of NETs (a process known as NETosis) and determine if frozen-thawed semen stimulates excessive NETosis.

Methods

  • Three different in vitro experiments were conducted which involved exposing neutrophils from female donkeys (known as Jenny PMNs) to fresh or frozen-thawed semen from male donkeys (known as jackass), isolated sperm, or seminal plasma (SP).
  • The researchers measured NET formation as a result of different treatments via manual quantification of stained slides.
  • A one-way analysis of variance (ANOVA), followed by a post hoc Sidak test, was used for statistical analysis.

Results

  • The release of NETs was found to depend on semen concentration and incubation time.
  • Frozen-thawed donkey sperm surprisingly did not show an increased NETosis rate relative to the control group. Conversely, fresh semen did stimulate a notably higher rate of NETosis.
  • NETosis also increased in the presence of SP, regardless if sperm was present or not. This was seen both with fresh and frozen samples.
  • The study also discovered that the presence of viable and motile sperm, even in the absence of SP, did not affect NETosis rates.

Conclusions

The researchers concluded that it is the seminal plasma in donkey semen, rather than any factors intrinsic to the sperm itself, that is able to trigger NETosis in a time- and semen concentration-dependent fashion. The exact physiological consequences of these findings in terms of the donkey’s response to semen remains unclear and requires further research.

Cite This Article

APA
Mateo-Otero Y, Zambrano F, Catalán J, Sánchez R, Yeste M, Miro J, Fernandez-Fuertes B. (2021). Seminal plasma, and not sperm, induces time and concentration-dependent neutrophil extracellular trap release in donkeys. Equine Vet J, 54(2), 415-426. https://doi.org/10.1111/evj.13457

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 54
Issue: 2
Pages: 415-426

Researcher Affiliations

Mateo-Otero, Yentel
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain.
Zambrano, Fabiola
  • Laboratory in Reproductive Medicine and Molecular Endocrinology, Center of Excellence in Translational Medicine-Scientific and Technological Bioresource Nucleus (CEMT - BIOREN), Universidad de La Frontera, Temuco, Chile.
  • Department of Preclinical Sciences, Faculty of Medicine, University of La Frontera, Temuco, Chile.
Catalán, Jaime
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain.
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, Barcelona, Spain.
Sánchez, Raúl
  • Laboratory in Reproductive Medicine and Molecular Endocrinology, Center of Excellence in Translational Medicine-Scientific and Technological Bioresource Nucleus (CEMT - BIOREN), Universidad de La Frontera, Temuco, Chile.
  • Department of Preclinical Sciences, Faculty of Medicine, University of La Frontera, Temuco, Chile.
Yeste, Marc
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain.
Miro, Jordi
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, Barcelona, Spain.
Fernandez-Fuertes, Beatriz
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, Girona, Spain.

MeSH Terms

  • Animals
  • Cryopreservation / veterinary
  • Equidae
  • Extracellular Traps
  • Female
  • Insemination, Artificial / veterinary
  • Male
  • Pregnancy
  • Semen
  • Semen Preservation / veterinary
  • Spermatozoa

Grant Funding

  • 2019-SGR-1229 / regional Government of Catalonia
  • 2020-FI-B-00412 / regional Government of Catalonia
  • AGL2017-88329-R / Ministry of Science and Innovation, Spain
  • RYC-2014-15581 / Ministry of Science and Innovation, Spain
  • 792212 / European Union's Horizon 2020 Research and Innovation Program

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Citations

This article has been cited 14 times.
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