88527 20210902121625.0 doi 10.3390/ijms21062093 sideral 117233 ART-2020-117233 eng Miguel-Jiménez, Sara Universidad de Zaragoza (orcid)0000-0001-6733-9373 Does melatonin exert its effect on ram sperm capacitation through nitric oxide synthase regulation? 2020 Access copy available to the general public Unrestricted Nitric oxide (NO·), synthesized from L-arginine by nitric oxide synthase (NOS), is involved in sperm functionality. NOS isoforms have been detected in spermatozoa from different species, and an increment in NOS activity during capacitation has been reported. This work aims to determine the presence and localization of NOS isoforms in ram spermatozoa and analyse their possible changes during in vitro capacitation. Likewise, we investigated the effect of melatonin on the expression and localization of NOS and NO· levels in capacitated ram spermatozoa. Western blot analysis revealed protein bands associated with neuronal NOS (nNOS) and epithelial NOS (eNOS) but not with inducible NOS (iNOS). However, the three isoforms were detected by indirect immunofluorescence (IFI), and their immunotypes varied over in vitro capacitation with cAMP-elevating agents. NO· levels (evaluated by DAF-2-DA/PI staining) increased after in vitro capacitation, and the presence of L-arginine in the capacitating medium raised NO· production and enhanced the acrosome reaction. Incubation in capacitating conditions with a high-cAMP medium with melatonin modified the NOS distribution evaluated by IFI, but no differences in Western blotting were observed. Melatonin did not alter NO· levels in capacitating conditions, so we could infer that its role in ram sperm capacitation would not be mediated through NO· metabolism. info:eu-repo/grantAgreement/ES/DGA/A07-17R info:eu-repo/grantAgreement/ES/MINECO/AGL2017-83799-R info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 5.923 2020 BIOCHEMISTRY & MOLECULAR BIOLOGY 67 / 297 = 0.226 2020 Q1 T1 CHEMISTRY, MULTIDISCIPLINARY 49 / 178 = 0.275 2020 Q2 T1 1.455 2020 Catalysis 2020 Q1 Computer Science Applications 2020 Q1 Inorganic Chemistry 2020 Q1 Spectroscopy 2020 Q1 Molecular Biology 2020 Q1 Organic Chemistry 2020 Q1 Physical and Theoretical Chemistry 2020 Q1 Medicine (miscellaneous) 2020 Q1 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Carvajal-Serna, Melissa Universidad de Zaragoza (orcid)0000-0003-3929-5064 Calvo, Silvia Universidad de Zaragoza Casao, Adriana Universidad de Zaragoza (orcid)0000-0003-1997-4262 Cebrián-Pérez, José Álvaro Universidad de Zaragoza (orcid)0000-0001-9842-6197 Muiño-Blanco, Teresa Universidad de Zaragoza (orcid)0000-0003-1306-8517 Pérez-Pe, Rosaura Universidad de Zaragoza (orcid)0000-0002-2312-6402 1002 819 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Zoología 1011 630 Universidad de Zaragoza Dpto. Microb.Ped.Radio.Sal.Pú. Área Microbiología 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 21, 6 (2020), 2093 [16 pp.] Int. j. mol. sci. International Journal of Molecular Sciences 1661-6596 1439897 http://zaguan.unizar.es/record/88527/files/texto_completo.pdf Versión publicada 479030 http://zaguan.unizar.es/record/88527/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:88527 articulos driver 2021-09-02-08:49:48 ARTICLE