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Cytosine methylation of sperm DNA in horse semen after cryopreservation

dc.contributor.authorAurich, Christine
dc.contributor.authorSchreiner, Bettina
dc.contributor.authorIlle, Natascha
dc.contributor.authorAlvarenga, Marco [UNESP]
dc.contributor.authorScarlet, Dragos
dc.contributor.institutionVetmeduni Vienna
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2018-12-11T17:03:02Z
dc.date.available2018-12-11T17:03:02Z
dc.date.issued2016-09-15
dc.description.abstractSemen processing may contribute to epigenetic changes in spermatozoa. We have therefore addressed changes in sperm DNA cytosine methylation induced by cryopreservation of stallion semen. The relative amount of 5-methylcytosine relative to the genomic cytosine content of sperm DNA was analyzed by ELISA. In experiment 1, raw semen (n = 6 stallions, one ejaculate each) was shock-frozen. Postthaw semen motility and membrane integrity were completely absent, whereas DNA methylation was similar in raw (0.4 ± 0.2%) and shock-frozen (0.3 ± 0.1%) semen (not significant). In experiment 2, three ejaculates per stallion (n = 6) were included. Semen quality and DNA methylation was assessed before addition of the freezing extender and after freezing-thawing with either Ghent (G) or BotuCrio (BC) extender. Semen motility, morphology, and membrane integrity were significantly reduced by cryopreservation but not influenced by the extender (e.g., total motility: G 69.5 ± 2.0, BC 68.4 ± 2.2%; P < 0.001 vs. centrifugation). Cryopreservation significantly (P < 0.01) increased the level of DNA methylation (before freezing 0.6 ± 0.1%, postthaw G 6.4 ± 3.7, BC 4.4 ± 1.5%; P < 0.01), but no differences between the freezing extenders were seen. The level of DNA methylation was not correlated to semen motility, morphology, or membrane integrity. The results demonstrate that semen processing for cryopreservation increases the DNA methylation level in stallion semen. We conclude that assessment of sperm DNA methylation allows for evaluation of an additional parameter characterizing semen quality. The lower fertility rates of mares after insemination with frozen-thawed semen may at least in part be explained by cytosine methylation of sperm-DNA induced by the cryopreservation procedure.en
dc.description.affiliationArtificial Insemination and Embryo Transfer Department for Small Animals and Horses Vetmeduni Vienna
dc.description.affiliationDepartment of Animal Reproduction & Veterinary Radiology Faculty of Veterinary Medicine FMVZ - Sao Paulo State University UNESP
dc.description.affiliationObstetrics Gynecology Andrology Department for Small Animals and Horses Vetmeduni Vienna
dc.description.affiliationUnespDepartment of Animal Reproduction & Veterinary Radiology Faculty of Veterinary Medicine FMVZ - Sao Paulo State University UNESP
dc.format.extent1347-1352
dc.identifierhttp://dx.doi.org/10.1016/j.theriogenology.2016.04.077
dc.identifier.citationTheriogenology, v. 86, n. 5, p. 1347-1352, 2016.
dc.identifier.doi10.1016/j.theriogenology.2016.04.077
dc.identifier.file2-s2.0-84970003153.pdf
dc.identifier.issn0093-691X
dc.identifier.scopus2-s2.0-84970003153
dc.identifier.urihttp://hdl.handle.net/11449/172995
dc.language.isoeng
dc.relation.ispartofTheriogenology
dc.rights.accessRightsAcesso aberto
dc.sourceScopus
dc.subjectCryopreservation
dc.subjectDNA methylation
dc.subjectHorse
dc.subjectSemen
dc.titleCytosine methylation of sperm DNA in horse semen after cryopreservationen
dc.typeArtigo
dspace.entity.typePublication
unesp.author.orcid0000-0001-6077-7362[1]
unesp.campusUniversidade Estadual Paulista (UNESP), Faculdade de Medicina Veterinária e Zootecnia, Botucatupt
unesp.departmentReprodução Animal e Radiologia Veterinária - FMVZpt

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