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Bradykinin-induced Ca2+ signaling in human subcutaneous fibroblasts involves ATP release via hemichannels leading to P2Y12 receptors activation

dc.contributor.authorPinheiro, Ana Rita
dc.contributor.authorParamos-de-Carvalho, Diogo
dc.contributor.authorCertal, Mariana
dc.contributor.authorCosta, Cristina
dc.contributor.authorMagalhães-Cardoso, Maria Teresa
dc.contributor.authorFerreirinha, Fátima
dc.contributor.authorCosta, M. Adelina
dc.contributor.authorCorreia-de-Sá, Paulo
dc.date.accessioned2014-01-22T12:15:24Z
dc.date.available2014-01-22T12:15:24Z
dc.date.issued2013
dc.description.abstractChronic musculoskeletal pain involves connective tissue remodeling triggered by inflammatory mediators, such as bradykinin. Fibroblast cells signaling involve changes in intracellular Ca2+ ([Ca2+]i). ATP has been related to connective tissue mechanotransduction, remodeling and chronic inflammatory pain, via P2 purinoceptors activation. Here, we investigated the involvement of ATP in bradykinin-induced Ca2+ signals in human subcutaneous fibroblasts. Bradykinin, via B2 receptors, caused an abrupt rise in [Ca2+]i to a peak that declined to a plateau, which concentration remained constant until washout. The plateau phase was absent in Ca2+-free medium; [Ca2+]i signal was substantially reduced after depleting intracellular Ca2+ stores with thapsigargin. Extracellular ATP inactivation with apyrase decreased the [Ca2+]i plateau. Human subcutaneous fibroblasts respond to bradykinin by releasing ATP via connexin and pannexin hemichannels, since blockade of connexins, with 2- octanol or carbenoxolone, and pannexin-1, with 10Panx, attenuated bradykinin-induced [Ca2+]i plateau, whereas inhibitors of vesicular exocytosis, such as brefeldin A and bafilomycin A1, were inactive. The kinetics of extracellular ATP catabolism favors ADP accumulation in human fibroblast cultures. Inhibition of ectonucleotidase activity and, thus, ADP formation from released ATP with POM-1 or by Mg2+ removal from media reduced bradykinin-induced [Ca2+]i plateau. Selective blockade of the ADP-sensitive P2Y12 receptor with AR-C66096 attenuated bradykinin [Ca2+]i plateau, whereas the P2Y1 and P2Y13 receptor antagonists, respectively MRS 2179 and MRS 2211, were inactive. Human fibroblasts exhibited immunoreactivity against connexin-43, pannexin-1 and P2Y12 receptor. Bradykinin induces ATP release from human subcutaneous fibroblasts via connexin and pannexin-1-containing hemichannels leading to [Ca2+]i mobilization through the cooperation of B2 and P2Y12 receptors.por
dc.identifier.doi10.1186/1478-811X-11-70pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.22/3435
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherBioMed Centralpor
dc.relation.ispartofseriesCell Communication and Signaling; Vol. 11
dc.relation.publisherversionhttp://www.biosignaling.com/content/11/1/70por
dc.subjectADPpor
dc.subjectATP releasepor
dc.subjectBradykininpor
dc.subjectCx43por
dc.subjectHemichannelspor
dc.subjectHuman subcutaneous fibroblastspor
dc.subjectPanx1por
dc.subjectP2 purinoceptorspor
dc.titleBradykinin-induced Ca2+ signaling in human subcutaneous fibroblasts involves ATP release via hemichannels leading to P2Y12 receptors activationpor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.titleCell Communication and Signalingpor
oaire.citation.volumeVol. 11por
person.familyNamePinheiro
person.givenNameAna Rita
person.identifier.orcid0000-0003-4310-7652
person.identifier.ridB-4018-2017
rcaap.rightsopenAccesspor
rcaap.typearticlepor
relation.isAuthorOfPublicationa99bce31-2fe7-4e36-af87-32d684d22ce0
relation.isAuthorOfPublication.latestForDiscoverya99bce31-2fe7-4e36-af87-32d684d22ce0

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