Endometrial explant culture for characterizing equine endometritis.
Abstract: Endometritis after insemination is ubiquitous in the horse and is associated with semen and/or bacteria in the uterus. In up to 40% of horses, inflammation persists causing infertility. An endometrial explant culture was developed to study uterine secretion of prostaglandin F(2alpha) (PGF(2alpha)) in response to physiological and pathological challenge. Methods: Uteri were collected from mares, the endometrium dissected and explants from the uterine body or horn cultured in William's or RPMI medium. The response of explants to oxytocin, semen or bacteria compared to untreated tissue was tested by collecting medium after 24 and 72 hr and measuring PGF(2alpha) by radioimmunoassay. Results: Explants from the uterine horn and cultured in William's medium secreted the most PGF(2alpha) after challenge with oxytocin. Explants treated with semen produced a PGF(2alpha) response after 72 hr. Explants collected from mares in the transition season treated with killed S. zooepidemicus or E. coli lipopolysaccharide (LPS) secreted increased concentrations of PGF(2alpha) after 24 and 72 hr. The response to LPS was inhibited by polymyxin B. Follicular and luteal phase explants did not respond to treatments. Conclusions: An endometrial explant culture was developed that measured PGF(2alpha) and may be used to study endometritis.
Publication Date: 2008-01-24 PubMed ID: 18211536DOI: 10.1111/j.1600-0897.2007.00548.xGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research study presents a new method to study equine endometritis, a common cause for infertility in horses. The researchers developed an endometrial explant culture model to gauge the uterine secretion of prostaglandin F(2alpha) (PGF(2alpha)) in response to various physiological and pathological challenges linked to endometritis.
Methods
- The researchers collected uteri from horses, separating the inner lining of the uterus (endometrium).
- Explants (tissue samples) were taken from either the body of the uterus or the uterine horn, and cultured in either William’s or RPMI medium.
- To stimulate the conditions of endometritis, the explants were treated with oxytocin, semen, or bacteria. The response was then assessed by monitoring the secretion of PGF(2alpha) at 24-hour and 72-hour intervals, using a technique called radioimmunoassay.
Results
- Explants from the uterine horn cultured in William’s medium secreted the highest amount of PGF(2alpha) when treated with oxytocin.
- Explants treated with semen showed a response after 72 hours, producing PGF(2alpha).
- Explants from mares, particularly during the transition season, secreted increased amounts of PGF(2alpha) when treated with S. zooepidemicus or E. coli (both implicated in endometritis). This response could be inhibited by polymyxin B, an antibiotic that binds to and neutralizes bacterial lipopolysaccharides.
- Importantly, explants in the follicular and luteal phases of the reproductive cycle showed no response to these treatments. This potentially indicates that the stage of the reproductive cycle affects a mare’s vulnerability to endometritis.
Conclusions
- A novel mechanism to study equine endometritis was developed with the use of endometrial explant culture. This method measures PGF(2alpha), a key biomarker in the inflammatory process linked to this disease.
- The model allows researchers to study the interactions between breeding factors (like semen), infectious agents (like bacteria), and host responses.
Cite This Article
APA
Nash D, Lane E, Herath S, Sheldon IM.
(2008).
Endometrial explant culture for characterizing equine endometritis.
Am J Reprod Immunol, 59(2), 105-117.
https://doi.org/10.1111/j.1600-0897.2007.00548.x Publication
Researcher Affiliations
- Department of Veterinary Clinical Sciences, The Royal Veterinary College, University of London, London, UK. dmn@aber.ac.uk
MeSH Terms
- Animals
- Anti-Bacterial Agents / pharmacology
- Dinoprost / metabolism
- Endometritis / blood
- Endometritis / physiopathology
- Endometritis / veterinary
- Endometrium / drug effects
- Endometrium / metabolism
- Estrous Cycle
- Female
- Histocytochemistry / veterinary
- Horse Diseases / blood
- Horse Diseases / pathology
- Horses
- Lipopolysaccharides / antagonists & inhibitors
- Lipopolysaccharides / pharmacology
- Oxytocin / pharmacology
- Polymyxin B / pharmacology
- Progesterone / blood
- Streptococcus equi
- Tissue Culture Techniques / methods
- Tissue Culture Techniques / veterinary
Citations
This article has been cited 10 times.- Rebordão MR, Amaral A, Fernandes C, Silva E, Lukasik K, Szóstek-Mioduchowska A, Pinto-Bravo P, Galvão A, Skarzynski DJ, Ferreira-Dias G. Enzymes Present in Neutrophil Extracellular Traps May Stimulate the Fibrogenic PGF(2α) Pathway in the Mare Endometrium. Animals (Basel) 2021 Sep 6;11(9).
- Monteiro de Barros MR, Davies-Morel MCG, Mur LAJ, Creevey CJ, Alison RH, Nash DM. Characterization of an Ex Vivo Equine Endometrial Tissue Culture Model Using Next-Generation RNA-Sequencing Technology. Animals (Basel) 2021 Jul 3;11(7).
- Ibrahim S, Hedia M, Taqi MO, Derbala MK, Mahmoud KGM, Ahmed Y, Ismail S, El-Belely M. Alterations in the Expression Profile of Serum miR-155, miR-223, miR-17, miR-200a, miR-205, as well as Levels of Interleukin 6, and Prostaglandins during Endometritis in Arabian Mares. Vet Sci 2021 Jun 4;8(6).
- Amaral A, Fernandes C, Szóstek-Mioduchowska A, Rebordão MR, Skarzynski DJ, Ferreira-Dias G. Noscapine Acts as a Protease Inhibitor of In Vitro Elastase-Induced Collagen Deposition in Equine Endometrium. Int J Mol Sci 2021 May 19;22(10).
- Amaral A, Fernandes C, Rebordão MR, Szóstek-Mioduchowska A, Lukasik K, Pinto-Bravo P, Telo da Gama L, Jan Skarzynski D, Ferreira-Dias G. Myeloperoxidase Inhibition Decreases the Expression of Collagen and Metallopeptidase in Mare Endometria under In Vitro Conditions. Animals (Basel) 2021 Jan 16;11(1).
- Amaral A, Fernandes C, Rebordão MR, Szóstek-Mioduchowska A, Lukasik K, Gawronska-Kozak B, Telo da Gama L, Skarzynski DJ, Ferreira-Dias G. The In Vitro Inhibitory Effect of Sivelestat on Elastase Induced Collagen and Metallopeptidase Expression in Equine Endometrium. Animals (Basel) 2020 May 16;10(5).
- Aboelsoued D, Toaleb NI, Abdel Megeed KN, Hassan SE, Ibrahim S. Cellular immune response and scanning electron microscopy in the evaluation of Moringa leaves aqueous extract effect on Cryptosporidium parvum in buffalo intestinal tissue explants. J Parasit Dis 2019 Sep;43(3):393-401.
- Nash DM, Paddison J, Davies Morel MCG, Barnea ER. Preimplantation factor modulates acute inflammatory responses of equine endometrium. Vet Med Sci 2018 Nov;4(4):351-356.
- Nash DM, Giles JL. Uterine inflammation and lessons from large animal models of endometritis. Nat Rev Immunol 2025 Dec;25(12):934-946.
- Köhne M, Hüsch R, Tönissen A, Schmidt M, Müsken M, Böttcher D, Hirnet J, Plötz M, Kittler S, Sieme H. Isolation and characterization of bacteriophages specific to Streptococcus equi subspecies zooepidemicus and evaluation of efficacy ex vivo. Front Microbiol 2024;15:1448958.
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