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Home / Equine Research Bank / Int J Mol Sci / Seminal Plasma, Sperm Concentration, and Sperm-PMN Interacti...
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.

References

This article includes 41 references
  1. Canisso IF, Panzani D, Miró J, Ellerbrock RE. Key Aspects of Donkey and Mule Reproduction. Vet. Clin. N. Am. Equine Pract. 2019;35:607–642.
    doi: 10.1016/j.cveq.2019.08.014pubmed: 31672204google scholar: lookup
  2. Quintero-Moreno A, Miró J, Teresa Rigau A, Rodríguez-Gil JE. Identification of sperm subpopulations with specific motility characteristics in stallion ejaculates. Theriogenology 2003;59:1973–1990.
    doi: 10.1016/S0093-691X(02)01297-9pubmed: 12600734google scholar: lookup
  3. Miró J, Lobo V, Quintero-Moreno A, Medrano A, Peña A, Rigau T. Sperm motility patterns and metabolism in Catalonian donkey semen. Theriogenology 2005;63:1706–1716.
    doi: 10.1016/j.theriogenology.2004.07.022pubmed: 15763113google scholar: lookup
  4. Flores E, Taberner E, Rivera MM, Peña A, Rigau T, Miró J, Rodríguez-Gil JE. Effects of freezing/thawing on motile sperm subpopulations of boar and donkey ejaculates. Theriogenology 2008;70:936–945.
    doi: 10.1016/j.theriogenology.2008.05.056pubmed: 18602684google scholar: lookup
  5. Rota A, Magelli C, Panzani D, Camillo F. Effect of extender, centrifugation and removal of seminal plasma on cooled-preserved Amiata donkey spermatozoa. Theriogenology 2008;69:176–185.
    doi: 10.1016/j.theriogenology.2007.09.003pubmed: 17945340google scholar: lookup
  6. Sabatini C, Mari G, Mislei B, Love C, Panzani D, Camillo F, Rota A. Effect of post-thaw addition of seminal plasma on motility, viability and chromatin integrity of cryopreserved donkey jack (Equus asinus) spermatozoa. Reprod. Domest. Anim. 2014;49:989–994.
    doi: 10.1111/rda.12419pubmed: 25256158google scholar: lookup
  7. Taberner E, Morató R, Mogas T, Miró J. Ability of Catalonian donkey sperm to penetrate zona pellucida-free bovine oocytes matured in vitro. Anim. Reprod. Sci. 2010;118:354–361.
    doi: 10.1016/j.anireprosci.2009.08.005pubmed: 19748750google scholar: lookup
  8. Vidament M, Vincent P, Martin FX, Magistrini M, Blesbois E. Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not. Anim. Reprod. Sci. 2009;112:22–35.
    doi: 10.1016/j.anireprosci.2008.03.016pubmed: 18502059google scholar: lookup
  9. De Oliveira JV, Oliveira PVDLF, E Oña CMM, Guasti PN, Monteiro GA, Da Silva YFRS, Papa PDM, Alvarenga MA, Junior JAD, Papa FO. Strategies to improve the fertility of fresh and frozen donkey semen. Theriogenology 2016;85:1267–1273.
    doi: 10.1016/j.theriogenology.2015.12.010pubmed: 26806444google scholar: lookup
  10. Kotilainen T, Huhtinen M, Katila T. Sperm-induced leukocytosis in the equine uterus. Theriogenology 1994;41:629–636.
    doi: 10.1016/0093-691X(94)90173-Gpubmed: 16727418google scholar: lookup
  11. Troedsson MHT, Loset K, Alghamdi AM, Dahms B, Crabo BG. Interaction between equine semen and the endometrium: The inflammatory response to semen. Anim. Reprod. Sci. 2001;68:273–278.
    doi: 10.1016/S0378-4320(01)00164-6pubmed: 11744271google scholar: lookup
  12. Rozeboom KJ, Rocha-Chavez G, Troedsson MH. Inhibition of neutrophil chemotaxis by pig seminal plasma in vitro: A potential method for modulating post-breeding inflammation in sows. Reproduction 2001;121:567–572.
    doi: 10.1530/rep.0.1210567pubmed: 11277876google scholar: lookup
  13. Matthijs A, Engel B, Woelders H. Neutrophil recruitment and phagocytosis of boar spermatozoa after artificial insemination of sows, and the effects of inseminate volume, sperm dose and specific additives in the extender. Reproduction 2003;125:357–367.
    doi: 10.1530/rep.0.1250357pubmed: 12611599google scholar: lookup
  14. Eisenbach M. Why are sperm cells phagocytosed by leukocytes in the female genital tract?. Med. Hypotheses 2003;60:590–592.
    doi: 10.1016/S0306-9877(03)00054-9pubmed: 12615529google scholar: lookup
  15. Görgens A, Leibold W, Klug E, Schuberth HJ, Martinsson G, Zerbe H. Inseminate components are modulating the chemotactic activity of uterine polymorphonuclear granulocytes (PMN) of mares. Anim. Reprod. Sci. 2005;89:308–310.
    pubmed: 16265752
  16. Vilés K, Rabanal R, Rodríguez-Prado M, Miró J. Influence of seminal plasma on leucocyte migration and amount of COX-2 protein in the jenny endometrium after insemination with frozen-thawed semen. Anim. Reprod. Sci. 2013;143:57–63.
    doi: 10.1016/j.anireprosci.2013.11.002pubmed: 24280633google scholar: lookup
  17. Miró J, Papas M. Post–Artificial Insemination Endometrial Inflammation and Its Control in Donkeys. J. Equine Vet. Sci. 2018;65:38–43.
    doi: 10.1016/j.jevs.2017.11.007google scholar: lookup
  18. Miró J, Taberner E, Rivera M, Peña A, Medrano A, Rigau T, Peñalba A. Effects of dilution and centrifugation on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen. Theriogenology 2009;72:1017–1022.
    doi: 10.1016/j.theriogenology.2009.06.012pubmed: 19747718google scholar: lookup
  19. Papas M, Arroyo L, Bassols A, Catalán J, Bonilla-Correal S, Gacem S, Yeste M, Miró J. Activities of antioxidant seminal plasma enzymes (SOD, CAT, GPX and GSR) are higher in jackasses than in stallions and are correlated with sperm motility in jackasses. Theriogenology 2019;140:180–187.
    doi: 10.1016/j.theriogenology.2019.08.032pubmed: 31479834google scholar: lookup
  20. Alghamdi AS, Lovaas BJ, Bird SL, Lamb GC, Rendahl AK, Taube PC, Foster DN. Species-specific interaction of seminal plasma on sperm-neutrophil binding. Anim. Reprod. Sci. 2009;114:331–344.
    doi: 10.1016/j.anireprosci.2008.10.015pubmed: 19081210google scholar: lookup
  21. Aloé S, Weber F, Behr B, Sauter-Louis C, Zerbe H. Modulatory effects of bovine seminal plasma on uterine inflammatory processes. Reprod. Domest. Anim. 2012;47:12–19.
    doi: 10.1111/j.1439-0531.2011.01792.xpubmed: 21535239google scholar: lookup
  22. Miró J, Vilés K, García W, Jordana J, Yeste M. Effect of donkey seminal plasma on sperm movement and sperm-polymorphonuclear neutrophils attachment in vitro. Anim. Reprod. Sci. 2013;140:164–172.
    doi: 10.1016/j.anireprosci.2013.06.007pubmed: 23891218google scholar: lookup
  23. Vilés K, Rabanal R, Rodríguez-Prado M, Miró J. Effect of ketoprofen treatment on the uterine inflammatory response after AI of jennies with frozen semen. Theriogenology 2013;79:1019–1026.
    doi: 10.1016/j.theriogenology.2013.01.006pubmed: 23453786google scholar: lookup
  24. Roth JA, Kaeberle ML. Isolation of neutrophils and eosinophils from the peripheral blood of cattle and comparison of their functional activities. J. Immunol. Methods 1981;45:153–164.
    doi: 10.1016/0022-1759(81)90209-Xpubmed: 7288194google scholar: lookup
  25. Siemsen DW, Schepetkin IA, Kirpotina LN, Lei B, Quinn MT. Neutrophil isolation from nonhuman species. Methods Mol. Biol. 2007;412:21–34.
    pubmed: 18453103
  26. Baumber J, Vo A, Sabeur K, Ball BA. Generation of reactive oxygen species by equine neutrophils and their effect on motility of equine spermatozoa. Theriogenology 2002;57:1025–1033.
    doi: 10.1016/S0093-691X(01)00710-5pubmed: 12041897google scholar: lookup
  27. Loftus JP, Williams JM, Belknap JK, Black SJ. In vivo priming and ex vivo activation of equine neutrophils in black walnut extract-induced equine laminitis is not attenuated by systemic lidocaine administration. Vet. Immunol. Immunopathol. 2010;138:60–69.
    doi: 10.1016/j.vetimm.2010.06.016pubmed: 20667603google scholar: lookup
  28. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, Weinrauch Y, Zychlinsky A. Neutrophil Extracellular Traps Kill Bacteria. Science 2004;303:1532–1535.
    doi: 10.1126/science.1092385pubmed: 15001782google scholar: lookup
  29. Fuchs TA, Abed U, Goosmann C, Hurwitz R, Schulze I, Wahn V, Weinrauch Y, Brinkmann V, Zychlinsky A. Novel cell death program leads to neutrophil extracellular traps. J. Cell Biol. 2007;176:231–241.
    doi: 10.1083/jcb.200606027pmc: PMC2063942pubmed: 17210947google scholar: lookup
  30. Kazzaz NM, Sule G, Knight JS. Intercellular interactions as regulators of netosis. Front. Immunol. 2016;7:453.
    doi: 10.3389/fimmu.2016.00453pmc: PMC5107827pubmed: 27895638google scholar: lookup
  31. Nakazawa D, Kumar SV, Desai J, Anders HJ. Neutrophil extracellular traps in tissue pathology. Histol. Histopathol. 2017;32:203–213.
    pubmed: 27593980
  32. Rebordão MR, Amaral A, Lukasik K, Szóstek-Mioduchowska A, Pinto-Bravo P, Galvão A, Skarzynski DJ, Ferreira-Dias G. Constituents of neutrophil extracellular traps induce in vitro collagen formation in mare endometrium. Theriogenology 2018;113:8–18.
    doi: 10.1016/j.theriogenology.2018.02.001pubmed: 29452855google scholar: lookup
  33. Branzk N, Lubojemska A, Hardison SE, Wang Q, Gutierrez MG, Brown GD, Papayannopoulos V. Neutrophils sense microbe size and selectively release neutrophil extracellular traps in response to large pathogens. Nat. Immunol. 2014;15:1017–1025.
    doi: 10.1038/ni.2987pmc: PMC4236687pubmed: 25217981google scholar: lookup
  34. Alghamdi AS, Foster DN. Seminal DNase Frees Spermatozoa Entangled in Neutrophil Extracellular Traps. Biol. Reprod. 2005;73:1174–1181.
    doi: 10.1095/biolreprod.105.045666pubmed: 16107606google scholar: lookup
  35. Alghamdi AS, Funnell BJ, Bird SL, Lamb GC, Rendahl AK, Taube PC, Foster DN. Comparative studies on bull and stallion seminal DNase activity and interaction with semen extender and spermatozoa. Anim. Reprod. Sci. 2010;121:249–258.
    doi: 10.1016/j.anireprosci.2010.06.003pubmed: 20638801google scholar: lookup
  36. Fichtner T, Kotarski F, Gärtner U, Conejeros I, Hermosilla C, Wrenzycki C, Taubert A. Bovine sperm samples induce different NET phenotypes in a NADPH oxidase-, PAD4-, and Ca++-dependent process. Biol. Reprod. 2020;102:902–914.
    doi: 10.1093/biolre/ioaa003pubmed: 31967293google scholar: lookup
  37. Schjenken JE, Robertson SA. The Female Response to Seminal Fluid. Physiol. Rev. 2020;100:1077–1117.
    doi: 10.1152/physrev.00013.2018pubmed: 31999507google scholar: lookup
  38. Menegazzi R, Decleva E, Dri P. Killing by neutrophil extracellular traps: Fact or folklore?. Blood 2012;119:1214–1216.
    doi: 10.1182/blood-2011-07-364604pubmed: 22210873google scholar: lookup
  39. Kenney RM. Minimal contamination techniques for breeding mares: Techniques and preliminary findings. Proc. Am. Assoc. Equine Pract. 1975;21:327–336.
  40. Bamba K. Evaluation of acrosomal integrity of boar spermatozoa by bright field microscopy using an eosin-nigrosin stain. Theriogenology 1988;29:1245–1251.
    doi: 10.1016/0093-691X(88)90004-0google scholar: lookup
  41. Palm F, Walter I, Budik S, Kolodziejek J, Nowotny N, Aurich C. Influence of different semen extenders and seminal plasma on PMN migration and on expression of IL-1β, IL-6, TNF-α and COX-2 mRNA in the equine endometrium. Theriogenology 2008;70:843–851.
    doi: 10.1016/j.theriogenology.2008.04.054pubmed: 18584861google scholar: lookup

Citations

This article has been cited 14 times.
  1. Gambini A, Smith JM, Gurkin RJ, Palacios PD. Current and Emerging Advanced Techniques for Breeding Donkeys and Mules. Animals (Basel) 2025 Mar 29;15(7).
    doi: 10.3390/ani15070990pubmed: 40218383google scholar: lookup
  2. Miró J, Martínez-Rodero I, Yeste M, Catalán J. Cryopreservation of Horse Sperm. Methods Mol Biol 2025;2897:193-206.
    doi: 10.1007/978-1-0716-4406-5_14pubmed: 40202637google scholar: lookup
  3. Fanelli D, Moroni R, Bocci C, Camillo F, Rota A, Panzani D. Interspecific and Intraspecific Artificial Insemination in Domestic Equids. Animals (Basel) 2023 Feb 7;13(4).
    doi: 10.3390/ani13040582pubmed: 36830369google scholar: lookup
  4. Gobato MLM, Segabinazzi LGTM, Scheeren VFC, Bandeira RS, Freitas-Dell'Aqua CP, Dell'Aqua JA Jr, Papa FO. Ability of donkey sperm to tolerate cooling: Effect of extender base and removal of seminal plasma on sperm parameters and fertility rates in mares. Front Vet Sci 2022;9:1011899.
    doi: 10.3389/fvets.2022.1011899pubmed: 36225802google scholar: lookup
  5. Catalán J, Yánez-Ortiz I, Tvarijonaviciute A, González-Arostegui LG, Rubio CP, Yeste M, Miró J, Barranco I. Impact of Seminal Plasma Antioxidants on Donkey Sperm Cryotolerance. Antioxidants (Basel) 2022 Feb 18;11(2).
    doi: 10.3390/antiox11020417pubmed: 35204299google scholar: lookup
  6. Morrell JM, Rocha A. A Novel Approach to Minimising Acute Equine Endometritis That May Help to Prevent the Development of the Chronic State. Front Vet Sci 2021;8:799619.
    doi: 10.3389/fvets.2021.799619pubmed: 35071389google scholar: lookup
  7. Yánez-Ortiz I, Catalán J, Delgado-Bermúdez A, Carluccio A, Miró J, Yeste M. Addition of Reduced Glutathione (GSH) to Freezing Medium Reduces Intracellular ROS Levels in Donkey Sperm. Vet Sci 2021 Dec 2;8(12).
    doi: 10.3390/vetsci8120302pubmed: 34941829google scholar: lookup
  8. Fantini P, Jiménez R, Vilés K, Iborra A, Palhares MS, Catalán J, Prades M, Miró J. Simple Tube Centrifugation Method for Platelet-Rich Plasma (PRP) Preparation in Catalonian Donkeys as a Treatment of Endometritis-Endometrosis. Animals (Basel) 2021 Oct 9;11(10).
    doi: 10.3390/ani11102918pubmed: 34679937google scholar: lookup
  9. Yánez-Ortiz I, Catalán J, Mateo-Otero Y, Dordas-Perpinyà M, Gacem S, Yeste N, Bassols A, Yeste M, Miró J. Extracellular Reactive Oxygen Species (ROS) Production in Fresh Donkey Sperm Exposed to Reductive Stress, Oxidative Stress and NETosis. Antioxidants (Basel) 2021 Aug 27;10(9).
    doi: 10.3390/antiox10091367pubmed: 34572999google scholar: lookup
  10. Miró J, Catalán J, Marín H, Yánez-Ortiz I, Yeste M. Specific Seminal Plasma Fractions Are Responsible for the Modulation of Sperm-PMN Binding in the Donkey. Animals (Basel) 2021 May 13;11(5).
    doi: 10.3390/ani11051388pubmed: 34068214google scholar: lookup
  11. Álvarez-Rodríguez M, Martinez-Pastor F. Molecular Determinants of Seminal Plasma on Sperm Biology and Fertility. Int J Mol Sci 2021 Mar 30;22(7).
    doi: 10.3390/ijms22073555pubmed: 33808064google scholar: lookup
  12. Catalán J, Papas M, Trujillo-Rojas L, Blanco-Prieto O, Bonilla-Correal S, Rodríguez-Gil JE, Miró J, Yeste M. Red LED Light Acts on the Mitochondrial Electron Chain of Donkey Sperm and Its Effects Depend on the Time of Exposure to Light. Front Cell Dev Biol 2020;8:588621.
    doi: 10.3389/fcell.2020.588621pubmed: 33365309google scholar: lookup
  13. Papas M, Catalán J, Recuero S, Morrell JM, Yeste M, Miró J. Single Layer Centrifugation Improves the Quality of Fresh Donkey Semen and Modifies the Sperm Ability to Interact with Polymorphonuclear Neutrophils. Animals (Basel) 2020 Nov 16;10(11).
    doi: 10.3390/ani10112128pubmed: 33207812google scholar: lookup
  14. Quartuccio M, Cristarella S, Medica P, Fazio E, Mazzullo G, Rifici C, Liotta L, Satué K. Endometrial Cytology During the Different Phases of the Estrous Cycle in Jennies: New Evidences. Animals (Basel) 2020 Jun 19;10(6).
    doi: 10.3390/ani10061062pubmed: 32575538google scholar: lookup
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