Characterization and identification of extracellular vesicles-coupled miRNA profiles in seminal plasma of fertile and subfertile rabbit bucks

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dc.contributor.authorSakr, Osama G.
dc.contributor.authorGad, Ahmed
dc.contributor.authorCañón Beltrán, Karina Esperanza
dc.contributor.authorCajas Suárez, Yulia Nathaly
dc.contributor.authorProchazka, Radek
dc.contributor.authorRizos, Dimitri
dc.contributor.authorRebollar, Pilar G.
dc.date.accessioned2024-05-27T15:46:46Z
dc.date.available2024-05-27T15:46:46Z
dc.date.issued2023-10-01
dc.description2023 Acuerdos transformativos CRUE
dc.description.abstractSeminal plasma (SP) provides essential nutrients, transport, and protection to the spermatozoa during their journey through the male and female reproductive tracts. Extracellular vesicles (EVs) are one of the main components of the SP with several biomolecular cargoes, including miRNAs, that can influence spermatozoa functions and interact with the cells of the female reproductive tract. This study aimed to isolate, characterize, and identify the miRNA expression profiles in the SP-EVs isolated from fertile (F) and subfertile (S) rabbit bucks that could serve as fertility biomarkers. In this study, the methods to isolate and identify EVs including exosomes, from SP of 3 F and S bucks have been developed. Ultracentrifugation and size exclusion chromatography analysis were using to isolate EVs from SP of F and S males that were qualitative and quantitively characterised using transmission electron microscopy, nanoparticle tracking analysis and western blotting. In addition, total RNA, including miRNA, was isolated, sequenced and identified from SP-EVs samples. Different SP-EVs concentrations (8.53 × 1011 ± 1.04 × 1011 and 1.84 × 1012 ± 1.75 × 1011 particles/mL of SP; P = 0.008), with a similar average size (143.9 ± 11.9 and 115.5 ± 2.4 nm; P = 0.7422) in F and S males, respectively was observed. Particle size was not significantly correlated with any kinetic parameter. The concentration of SP-EVs was positively correlated with the percentage of abnormal forms (r = 0.94; P < 0.05) and with the percentage of immotile spermatozoa (r = 0.88; P < 0.05). Small-RNA-seq analysis identified a total of 267 and 244 expressed miRNAs in the F and S groups, respectively. Two miRNAs (let-7b-5p and let-7a-5p) were the top most abundant miRNAs in both groups. Differential expression analysis revealed that 9 miRNAs including miR-190b-5p, miR-193b-5p, let-7b-3p, and miR-378–3p, and another 9 miRNAs including miR-7a-5p, miR-33a-5p, miR-449a-5p, and miR-146a-5p were significantly up- and downregulated in the F compared to the S group, respectively. The SP from F and S rabbit males contains EVs with different miRNA cargo correlated with spermatogenesis, homeostasis, and infertility, which could be used as biomarkers for male fertility and potential therapies for assisted reproductive technologies.
dc.description.departmentSección Deptal. de Bioquímica y Biología Molecular (Veterinaria)
dc.description.facultyFac. de Veterinaria
dc.description.fundingtypeAPC financiada por la UCM
dc.description.refereedTRUE
dc.description.sponsorshipEuropean Commission
dc.description.sponsorshipMinisterio de Ciencia e Innovación (España)
dc.description.statuspub
dc.identifier.citationCharacterization and identification of extracellular vesicles-coupled miRNA profiles in seminal plasma of fertile and subfertile rabbit bucks, Theriogenology, Volume 209, 2023, Pages 76-88, https://doi.org/10.1016/j.theriogenology.2023.06.020
dc.identifier.doi10.1016/j.theriogenology.2023.06.020
dc.identifier.essn1879-3231
dc.identifier.issn0093-691X
dc.identifier.officialurlhttps://doi.org/10.1016/j.theriogenology.2023.06.020
dc.identifier.urihttps://hdl.handle.net/20.500.14352/104465
dc.journal.titleTheriogenology
dc.language.isoeng
dc.page.final88
dc.page.initial76
dc.publisherElsevier
dc.relation.projectIDCZ.02.1.01/0.0/0.0/15_003/0000460
dc.relation.projectIDinfo:eu-repo/grantAgreement/MICIIN//RTI 2018-094404-B-C-21
dc.relation.projectIDinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-111641RB-I00/ES/VESICULAS EXTRACELULARES: MODULADORES DE LA INTERACCION MATERNO-EMBRIONARIA Y DE LA FERTILIDAD EN BOVINO/
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 Internationalen
dc.rights.accessRightsopen access
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.cdu577.1/2
dc.subject.keywordExtracellular vesicles
dc.subject.keywordMale fertility
dc.subject.keywordmiRNA
dc.subject.keywordSeminal quality
dc.subject.keywordSeminal plasma
dc.subject.keywordRabbit
dc.subject.ucmMicrobiología (Veterinaria)
dc.subject.unesco3109 Ciencias Veterinarias
dc.titleCharacterization and identification of extracellular vesicles-coupled miRNA profiles in seminal plasma of fertile and subfertile rabbit bucks
dc.typejournal article
dc.type.hasVersionVoR
dc.volume.number209
dspace.entity.typePublication
relation.isAuthorOfPublication252147fa-6c76-4c5d-b884-af3fa2c738c3
relation.isAuthorOfPublicationf4da9b8e-f83c-49ec-adff-fafcd0bf3ef7
relation.isAuthorOfPublication.latestForDiscovery252147fa-6c76-4c5d-b884-af3fa2c738c3
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