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Isolation and characterization of extracellular vesicle subsets in donkey seminal plasma

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dc.contributor.authorCatalán, Jaime
dc.contributor.authorMartínez Díaz, Pablo
dc.contributor.authorParra, Ana
dc.contributor.authorBonet, Sergi
dc.contributor.authorYeste, Marc
dc.contributor.authorRoca, Jordi
dc.contributor.authorBarranco Cascales, Isabel
dc.contributor.authorMiró, Jordi
dc.contributor.departmentMedicina y Cirugía Animal
dc.contributor.otherFacultad de Veterinaria
dc.date.accessioned2025-10-17T07:36:45Z
dc.date.available2025-10-17T07:36:45Z
dc.date.copyright© 2025 The Authors
dc.date.issued2025-05-22
dc.description.abstractSeminal plasma (SP), a fluid composed of secretions from the male genital tract, is rich in seminal extracellular vesicles (sEVs), nano-sized particles surrounded by a lipid bilayer membrane and loaded with functionally active molecules. Seminal EVs are secreted by functional cells of the male genital tract and play a key role in modulating reproductive processes, including sperm function and immune response in the female genital tract. The aim of this study was to isolate and characterize sEVs from donkey SP for the first time. Nine SP samples were collected from nine healthy and reproductive active donkeys. The SP samples were randomly pooled to create three pools (three SP samples per pool). The SP pools were subjected to differential centrifugation and size-exclusion chromatography to separately isolate two subsets of sEVs: small (S-) and large (L-). Orthogonal characterization of sEV samples was performed according to MISEV 2023 guidelines, including morphology (by cryogenic electron microscopy), concentration (by total protein concentration and total and CFSE positive particles by flow cytometry [FC]), particle size distribution (by dynamic light scattering), purity (by albumin assessment by FC), and specific EV protein markers (tetraspanins CD9, CD63, and CD81, and HSP70 by FC). The results showed that donkey SP is highly enriched in sEVs. Size differences were found between both sEV subsets, with S-sEVs being smaller (∼160 nm) and L-sEVs larger (∼290 nm). Both sEV subsets were positive for the four EV protein markers. However, the percentage of CD81-positive events was higher in S-sEV samples than in L-sEV samples (P < 0.05). This study is the first to isolate and characterize sEVs in donkey SP, demonstrating their heterogeneity and suggesting differences in biogenesis and function between S-sEVs and L-sEVs.
dc.formatapplication/pdf
dc.format.extent9
dc.identifier.citationTheriogenology 244 (2025) 117501
dc.identifier.doihttps://doi.org/10.1016/j.theriogenology.2025.117501
dc.identifier.eissn1879-3231
dc.identifier.issn0093-691X
dc.identifier.urihttp://hdl.handle.net/10201/167369
dc.languageeng
dc.publisherElsevier
dc.relationThe study was funded by the following grants: (1) JDC2022-049684-I funded by (Ministry of Science, Innovation and Universities, Spain [MICIU]) MICIU/AEI/10.13039/501100011033 and European Union NextGenerationEU/PRTR; (2) PID2020-113493RB-I00 funded by MICIU/AEI/10.13039/501100011033; (3) PID2022-137738NA-I00 funded by MICIU/AEI/10.13039/501100011033 and ERDF/EU; (4) RYC2021-034546-I funded by MICIU/AEI/10.13039/501100011033 and European Union NextGenerationEU/PRTR; (5) 21935/PI/22 and (6) 22258/PDC/23 funded by the Seneca Foundation (Murcia, Spain). (7) PID2020-113320RB-I00) funded by MICIU/AEI/10.13039/501100011033 and (8) 2017-SGR-1229 and 2021-SGR-00900 funded by Catalan Agency for the Management of University and Research Grants, Regional Government of Catalonia, Spain.
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S0093691X25002274?via%3Dihub
dc.rightsAttribution 4.0 Internacional
dc.rights.accessRightsinfo:eu-repo/semantics/openAccess
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectDonkey
dc.subjectExtracellular vesicles
dc.subjectSeminal plasma
dc.subjectSize-exclusion chromatography
dc.subject.odsNo relacionado con ningún objetivo de desarrollo sostenible
dc.titleIsolation and characterization of extracellular vesicle subsets in donkey seminal plasma
dc.typeinfo:eu-repo/semantics/article
dspace.entity.typePublicationes
relation.isAuthorOfPublication57ff06c2-c0ce-46eb-b327-b84f76532b8e
relation.isAuthorOfPublication3c34d0d5-7d7d-4bcc-a119-8906f53baa7b
relation.isAuthorOfPublication.latestForDiscovery57ff06c2-c0ce-46eb-b327-b84f76532b8e
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