000148919 001__ 148919 000148919 005__ 20250123152145.0 000148919 0247_ $$2doi$$a10.3389/fcell.2021.655794 000148919 0248_ $$2sideral$$a126600 000148919 037__ $$aART-2021-126600 000148919 041__ $$aeng 000148919 100__ $$0(orcid)0000-0001-6733-9373$$aMiguel-Jiménez S.$$uUniversidad de Zaragoza 000148919 245__ $$aNADPH Oxidase 5 and Melatonin: Involvement in Ram Sperm Capacitation 000148919 260__ $$c2021 000148919 5060_ $$aAccess copy available to the general public$$fUnrestricted 000148919 5203_ $$aReactive oxygen species (ROS) play an essential role in mammalian sperm capacitation. NADPH oxidase 5 (NOX5) has been described as the main source of ROS production in some mammalian spermatozoa, such as human and equine. On the other hand, melatonin can decrease cellular ROS levels and regulates NOX activity in somatic cells. Therefore, the objectives of this work were (1) to identify NOX5 in ram spermatozoa and analyze its possible changes during in vitro capacitation and (2) to investigate the effect of melatonin on NOX5 expression and localization and on superoxide levels in capacitated ram spermatozoa. Protein bands associated with NOX5 were detected by Western blot analysis. Likewise, indirect immunofluorescence (IIF) revealed six different immunotypes for NOX5, which varied throughout in vitro capacitation. Superoxide (O2·–), evaluated by DHE/Yo-Pro-1, rose after in vitro capacitation and in the presence of the calcium ionophore A23187 but decreased in the presence of the NOX inhibitor GKT136901. GKT also reduced the percentage of capacitated and acrosome-reacted spermatozoa that had increased during incubation in capacitating conditions. The presence of melatonin at micromolar concentrations avoided the increment in O2·– and the changes in NOX5 immunotypes provoked by capacitation. In conclusion, NOX5 is present in ram spermatozoa and the changes in its distribution, associated with sperm capacitation, can be prevented by melatonin. To this extent, it could imply that melatonin exerts its antioxidant role, at least in part, by modulating NOX5 activity during ram sperm capacitation. © Copyright © 2021 Miguel-Jiménez, Pina-Beltrán, Gimeno-Martos, Carvajal-Serna, Casao and Pérez-Pe. 000148919 536__ $$9info:eu-repo/grantAgreement/ES/DGA/A07-17R$$9info:eu-repo/grantAgreement/ES/MINECO/AGL2017-83799-R 000148919 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/ 000148919 590__ $$a6.081$$b2021 000148919 591__ $$aDEVELOPMENTAL BIOLOGY$$b6 / 39 = 0.154$$c2021$$dQ1$$eT1 000148919 591__ $$aCELL BIOLOGY$$b68 / 195 = 0.349$$c2021$$dQ2$$eT2 000148919 592__ $$a1.44$$b2021 000148919 593__ $$aDevelopmental Biology$$c2021$$dQ1 000148919 593__ $$aCell Biology$$c2021$$dQ1 000148919 594__ $$a3.5$$b2021 000148919 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion 000148919 700__ $$aPina-Beltrán B. 000148919 700__ $$0(orcid)0000-0002-1685-6845$$aGimeno-Martos S. 000148919 700__ $$0(orcid)0000-0003-3929-5064$$aCarvajal-Serna M.$$uUniversidad de Zaragoza 000148919 700__ $$0(orcid)0000-0003-1997-4262$$aCasao A.$$uUniversidad de Zaragoza 000148919 700__ $$0(orcid)0000-0002-2312-6402$$aPérez-Pe R.$$uUniversidad de Zaragoza 000148919 7102_ $$11002$$2819$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Zoología 000148919 773__ $$g9 (2021), [14 pp]$$tFrontiers in Cell and Developmental Biology$$x2296-634X 000148919 787__ $$tDatos asociados$$tMaterial suplementario$$whttps://doi.org/10.6084/m9.figshare.13603226$$whttps://doi.org/10.6084/m9.figshare.13606394 000148919 8564_ $$s4011947$$uhttps://zaguan.unizar.es/record/148919/files/texto_completo.pdf$$yVersión publicada 000148919 8564_ $$s2219565$$uhttps://zaguan.unizar.es/record/148919/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada 000148919 909CO $$ooai:zaguan.unizar.es:148919$$particulos$$pdriver 000148919 951__ $$a2025-01-23-14:47:05 000148919 980__ $$aARTICLE