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dc.contributor.authorNarváez-Peláez, Manuel 
dc.contributor.authorBorroto Escuela, Dasiel Óscar
dc.contributor.authorMillón-Peñuela, Carmelo 
dc.contributor.authorFlores-Burgess, Antonio 
dc.contributor.authorGago-Calderón, Belén 
dc.contributor.authorSantín-Núñez, Luis Javier 
dc.contributor.authorFuxe, Kjell
dc.contributor.authorNarváez-Bueno, José Ángel 
dc.contributor.authorDíaz-Cabiale, Zaida 
dc.date.accessioned2015-10-05T12:24:18Z
dc.date.available2015-10-05T12:24:18Z
dc.date.created2015-10-05
dc.date.issued2015-10-05
dc.identifier.urihttp://hdl.handle.net/10630/10443
dc.description.abstractWe have recently described a Galanin receptor 2(GALR2) and Neuropeptide Y Y1 receptor(NPYY1R) interaction at behavioural, cellular and receptor levels through GALR2/NPYY1R heterodimers. The aim of this work was to study if GALR2 and NPYY1R costimulation modified NPYY1R internalization and β-Arrestin recruitment after in HEK293T cells. HEK293T cells were transfected with NPYY1REGFPor β-Arrestin2GFP2 cloned with standard molecular biology techniques employing PCR and fragment replacement strategies. NPYY1REGFP/GALR2 and NPYY1R/GALR2 with β- Arrestin2GFP2 HEK293T coexpressing cells were incubated with NPY 1μM and/or GAL1μM, at different times. Antagonist studies were performed 15 min prior to the addition of agonist with NPYY1R antagonist BIBP3226 10μM or GALR2 antagonist M871 10 μM. Timed-interval images of NPYY1REGFP or β-Arrestin2GFP2 endosomes in different cell groups were acquired using a confocal microscope following agonist addition. Percentage of internalization was determined by Leica software analysis of total membrane fluorescence compared to total internal compartment fluorescence at the various time points. We observed that addition of NPY induced a rapid decrease in the cell surface expression of NPYY1REGFP and a redistribution of β-Arrestin2GFP2. In fact, we observed a maximum of internalization of 80% three minutes after the NPY stimulation. However, combined treatment with GAL and NPY induced a delay in the internalization of NPYY1REGFP, with a maximum of internalization thirty minutes after the co-stimulation. Moreover, a delay in the β-Arrestin2GFP2 redistribution was observed. The specific GALR2 antagonist M871 abolished these delays in internalization of NPYY1REGFP and β-Arrestin2GFP2 redistribution, suggesting that this effect was mediated through the coactivation of GALR2 and NPYY1R. These results demonstrate that costimulation with GAL and NPY delays the internalization of  NPYY1REGFP by decreasing recruitment of β-Arrestin2GFP2 and probably could change intracellular signaling. This study was supported by Junta de Andalucia CVI6476.es_ES
dc.description.sponsorshipJunta de Andalucia CVI6476.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech.es_ES
dc.language.isoenges_ES
dc.rightsinfo:eu-repo/semantics/openAccesses_ES
dc.subjectNeuropéptidoses_ES
dc.subject.otherAnsiedades_ES
dc.subject.otherNeuropéptidoes_ES
dc.subject.otherInteracción receptoreses_ES
dc.subject.otherGalaninaes_ES
dc.titleGalanin receptor 2 modifies neuropeptide Y Y1 receptor internalization and β-Arrestin recruitmentes_ES
dc.typeinfo:eu-repo/semantics/conferenceObjectes_ES
dc.centroFacultad de Medicinaes_ES
dc.relation.eventtitle16 Congreso de la SENCes_ES
dc.relation.eventplaceGranadaes_ES
dc.relation.eventdate09-2015es_ES
dc.identifier.orcidhttp://orcid.org/0000-0003-0922-4900es_ES
dc.rights.ccby-nc-nd


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