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International journal of molecular sciences2020; 21(10); doi: 10.3390/ijms21103478

Seminal Plasma, Sperm Concentration, and Sperm-PMN Interaction in the Donkey: An In Vitro Model to Study Endometrial Inflammation at Post-Insemination.

Abstract: In the donkey, artificial insemination (AI) with frozen-thawed semen is associated with low fertility rates, which could be partially augmented through adding seminal plasma (SP) and increasing sperm concentration. On the other hand, post-AI endometrial inflammation in the jenny is significantly higher than in the mare. While previous studies analyzed this response through recovering Polymorphonuclear Neutrophils (PMN) from uterine washings, successive lavages can detrimentally impact the endometrium, leading to fertility issues. For this reason, the first set of experiments in this work intended to set an in vitro model through harvesting PMN from the peripheral blood of jennies. Thereafter, how PMN, which require a triggering agent like formyl-methionyl-leucyl-phenylalanine (FMLP) to be activated, are affected by donkey semen was interrogated. Finally, we tested how four concentrations of spermatozoa (100 × 106, 200 × 106, 500 × 106 and 1000 × 106 spermatozoa/mL) affected their interaction with PMN. We observed that semen, which consists of sperm and SP, is able to activate PMN. Whereas there was a reduced percentage of spermatozoa phagocytosed by PMN, most remained attached on the PMN surface or into a surrounding halo. Spermatozoa not attached to PMN were viable, and most of those bound to PMN were also viable and showed high tail beating. Finally, only sperm concentrations higher than 500 × 106 spermatozoa/mL showed free sperm cells after 3 h of incubation, and percentages of spermatozoa not attached to PMN were higher at 3 h than at 1 h, exhibiting high motility. We can thus conclude that semen activates PMN in the donkey, and that the percentage of spermatozoa phagocytosed by PMN is low. Furthermore, because percentages of spermatozoa not attached to PMN were higher after 3 h than after 1 h of incubation, we suggest that PMN-sperm interaction plays an instrumental role in the reproductive strategy of the donkey.
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Publication Date: 2020-05-14 PubMed ID: 32423134PubMed Central: PMC7278951DOI: 10.3390/ijms21103478Google 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.

This study explores the causes behind the low fertility rates in donkeys when artificial insemination (AI) with frozen-thawed semen is used. It finds that endometrial inflammation in female donkeys (jennies) post-AI is potentially linked to biological functions involving neuthrophils (PMN), specific immune cells. Notably, increased sperm concentrations and seminal plasma may improve fertility rates.

Understanding the Research Context

  • The research seeks to address the issue of low fertility rates in donkeys when artificial insemination (AI) is used with frozen-thawed semen. This technique poses a challenge due to the high levels of endometrial inflammation seen in female donkeys (jennies) after insemination.
  • This inflammation is believed to be linked to Polymorphonuclear Neutrophils (PMN), immune cells that could be involved in fertility issues. Previous attempts to examine this interaction have potentially led to damaging the endometrium further, as the procedure involves invasive methods like uterine washings to obtain PMN cells.

Experimenting with an In Vitro Model

  • In order to find a new approach to study this, the researchers developed an in vitro model, whereby PMN cells are obtained from the peripheral blood of jennies.
  • In typical situations, these PMN cells require an activating agent, in this case formyl-methionyl-leucyl-phenylalanine (FMLP), to function. Therefore, the reaction of donkey semen with these cells was tested.
  • This study examined various concentrations of sperm (100 × 10, 200 × 10, 500 × 10, and 1000 × 10 spermatozoa/mL)to see how they affected the interaction with PMN cells.

Findings

  • Semen was found to activate PMN cells, although a reduced percentage of spermatozoa were engulfed (phagocytosed) by them. Most spermatozoa remained on the PMN’s surface or in a surrounding area (halo).
  • Interestingly, even after PMN interaction, most spermatozoa remained viable. Notably, those remaining spermatozoa not attached to PMN exhibited strong motility.
  • Only higher concentrations of sperm (500 × 10 spermatozoa/mL upwards) showed free sperm cells after 3 hours of incubation.
  • Importantly, the percentage of non-attached spermatozoa increased after 3 hours of incubation compared to the 1 hour mark, suggesting that the PMN-sperm interaction may play an important role in the reproduction strategy of donkeys.

Conclusion

  • The study concludes that semen activates PMN cells in donkeys and that there is a low rate of engulfment of spermatozoa by PMN cells.
  • The interaction between sperm and PMN appears to be crucial for the reproductive strategy due to an observed increase in the number of free-moving sperm over time.
  • This study could lead to potential improvements in artificial insemination techniques for donkeys, especially regarding the use of frozen-thawed semen.

Cite This Article

APA
Miró J, Marín H, Catalán J, Papas M, Gacem S, Yeste M. (2020). Seminal Plasma, Sperm Concentration, and Sperm-PMN Interaction in the Donkey: An In Vitro Model to Study Endometrial Inflammation at Post-Insemination. Int J Mol Sci, 21(10). https://doi.org/10.3390/ijms21103478

Publication

International journal of molecular sciences
ISSN: 1422-0067
NlmUniqueID: 101092791
Country: Switzerland
Language: English
Volume: 21
Issue: 10

Researcher Affiliations

Miró, Jordi
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra (Cerdanyola del Vallès), Spain.
Marín, Henar
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra (Cerdanyola del Vallès), Spain.
Catalán, Jaime
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra (Cerdanyola del Vallès), Spain.
Papas, Marion
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra (Cerdanyola del Vallès), Spain.
Gacem, Sabrina
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra (Cerdanyola del Vallès), Spain.
Yeste, Marc
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, E-17003 Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, E-17003 Girona, Spain.

MeSH Terms

  • Animals
  • Cryopreservation
  • Endometritis / metabolism
  • Endometritis / pathology
  • Endometrium / metabolism
  • Endometrium / pathology
  • Equidae / psychology
  • Female
  • Inflammation / blood
  • Inflammation / genetics
  • Inflammation / pathology
  • Insemination, Artificial
  • Male
  • Neutrophils / metabolism
  • Neutrophils / pathology
  • Semen / metabolism
  • Sperm Motility / genetics
  • Spermatozoa / metabolism
  • Spermatozoa / pathology
  • Uterus / metabolism
  • Uterus / pathology

Grant Funding

  • RYC-2014-1558 & AGL2017-88329R / 2017-SGR-1229 / 2017/72180128 / This research was supported by the Ministry of Science, Innovation and Universities, Spain (Grants: RYC-2014-15581 and AGL2017- 88329-R), and the Regional Government of Catalonia (2017-SGR-1229). J.C. was funded by the National Commission of Scientific Re

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

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