Multiparameter Flow Cytometry Assay for Analysis of Nitrosative Stress Status in Human Spermatozoa

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datacite.alternateIdentifier.citationCytometry Part A, 97 (12), 1247-1238, 2020
datacite.alternateIdentifier.doi10.1002/cyto.a.24170
datacite.alternateIdentifier.issn1552-4922
datacite.creatorUribe, Pamela
datacite.creatorMeriño, Juan
datacite.creatorManquemilla, Emilio
datacite.creatorVillagrán, Camila
datacite.creatorVega, Etelinda
datacite.creatorZambrano, Fabiola
datacite.creatorSchulz, Mabel A.
datacite.creatorPezo, Felipe
datacite.creatorVillegas, Juana V.
datacite.creatorBoguen, R.
datacite.date2020
datacite.rightsRegistro bibliográfico
datacite.subjectHuman Sperm
datacite.subjectMitochondria
datacite.subjectMultiparametric Flow Cytometry
datacite.subjectReactive Nitrogen Species
datacite.subjectSperm Function
datacite.subjectCryopreservation
datacite.subjectFlow Cytometry
datacite.subjectHuman
datacite.subjectMale
datacite.subjectMetabolism
datacite.subjectMitochondrial Membrane Potential
datacite.subjectNitrosative Stress
datacite.subjectSpermatozoon
datacite.subjectCryopreservation
datacite.subjectFlow Cytometry
datacite.subjectHumans
datacite.subjectMale
datacite.subjectMembrane Potential, Mitochondrial
datacite.subjectNitrosative Stress
datacite.subjectSpermatozoa
datacite.titleMultiparameter Flow Cytometry Assay for Analysis of Nitrosative Stress Status in Human Spermatozoa
dc.contributor.authorBOGUEN OJEDA, RODRIGO ESTEBAN
dc.description.abstractMale infertility is an increasing health problem, and oxidative/nitrosative stress plays an important role in the etiology of this condition. Nitrosative stress due to excessive levels of reactive nitrogen species (RNS) is associated with impaired male fertility. Flow cytometry may be a useful tool for semen evaluation, but the availability of multiparameter assays for analysis of sperm quality is limited. The present study standardized a multiparameter flow cytometry analysis for nitrosative stress status in human spermatozoa in a single assay. A suitable multicolor fluorochrome panel was designed and consisted of fluorescein-boronate to detect peroxynitrite, a highly RNS, propidium iodide to analyze viability, tetramethylrhodamine methyl ester perchlorate to detect mitochondrial membrane potential (MMP) and monobromobimane to analyze thiol oxidation. Proper positive and negative controls for each fluorochrome were used to establish the technique, and sperm cells of different qualities and spermatozoa subjected to cryopreservation were analyzed. The results showed that the controls clearly discriminated between the high and low fluorescence intensities for each fluorochrome. The analysis of sperm cells of different quality demonstrated that the assay properly detected differences in all parameters analyzed according to sperm quality. The results may be reported as the mean fluorescence intensity of each fluorochrome and the percentage of cells exhibiting different characteristics. In conclusion, a protocol was standardized to analyze nitrosative stress status, including peroxynitrite production, viability, MMP, and thiol oxidation, in a single analysis using flow cytometry. This protocol may be applied to research approaches and clinical andrology to improve the evaluation of sperm quality and provide a promising tool to increase the use of clinical flow cytometry. © 2020 International Society for Advancement of Cytometry. © 2020 Elsevier B.V., All rights reserved.
dc.description.ia_keywordcytometry, nitrosative, stress, flow, sperm, quality, fluorochrome
dc.identifier.issn1552-4930
dc.identifier.urihttps://repositoriodigital.uct.cl/handle/10925/3074
dc.language.isoen
dc.publisherWiley
dc.relationinstname: ANID
dc.relationreponame: Repositorio Digital RI2.0
dc.rights.driverinfo:eu-repo/semantics/openAccess
dc.sourceCytometry Part A
dc.subject.ia_odsODS 3: Salud y bienestar
dc.subject.ia_oecd1nCiencias Naturales
dc.subject.ia_oecd2nCiencias Biológicas
dc.subject.ia_oecd3nBiología General
dc.type.driverinfo:eu-repo/semantics/article
dc.type.driverhttp://purl.org/coar/resource_type/c_2df8fbb1
dc.type.openaireinfo:eu-repo/semantics/publishedVersion
dspace.entity.typePublication
oaire.citationEdition2020
oaire.citationEndPage1247
oaire.citationIssue12
oaire.citationStartPage1238
oaire.citationTitleCytometry Part A
oaire.citationVolume97
oaire.fundingReferenceCONICYT ANID PAI 79160030
oaire.fundingReferenceANID FONDECYT 11170758 (Regular)
oaire.fundingReferenceUniversidad de La Frontera (Dirección de Investigación, VRIP)
oaire.licenseConditionCopyright © International Society for Advancement of Cytometry, 2020
oaire.resourceTypeArtículo
oaire.resourceType.enArticle
relation.isAuthorOfPublicationc9b8540f-e025-4d57-95dd-0de22ec54fa0
relation.isAuthorOfPublication.latestForDiscoveryc9b8540f-e025-4d57-95dd-0de22ec54fa0
uct.catalogadorjvu
uct.comunidadCiencias de la Saluden_US
uct.departamentoDepartamento de Procesos Diagnósticos y Evaluación
uct.facultadFacultad de Ciencias de la Salud
uct.indizacionScience Citation Index Expanded - SCIE
uct.indizacionScopus
uct.indizacionPubMed
uct.indizacionEMBASE
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