dc.creatorEspelt, Maria Victoria
dc.creatorde Tezanos Pinto, Felicitas
dc.creatorAlvarez, Cora Lilia
dc.creatorSanchez Alberti, German
dc.creatorIncicco, Juan Jeremías
dc.creatorLeal Denis, Maria Florencia
dc.creatorDavio, Carlos Alberto
dc.creatorSchwarzbaum, Pablo Julio
dc.date.accessioned2017-06-13T17:51:29Z
dc.date.available2017-06-13T17:51:29Z
dc.date.created2017-06-13T17:51:29Z
dc.date.issued2013-03
dc.identifierEspelt, Maria Victoria; de Tezanos Pinto, Felicitas; Alvarez, Cora Lilia; Sanchez Alberti, German; Incicco, Juan Jeremías; et al.; On the role of ATP release, ectoATPase activity, and extracellular ADP in the regulatory volume decrease of Huh-7 human hepatoma cells; American Physiological Society; American Journal Of Physiology-cell Physiology; 304; 10; 3-2013; 1013-1026
dc.identifier0363-6143
dc.identifierhttp://hdl.handle.net/11336/18105
dc.identifierCONICET Digital
dc.identifierCONICET
dc.description.abstractHypotonicity triggered in human hepatoma cells (Huh-7) the release of ATP and cell swelling, followed by volume regulatory decrease (RVD). We analyzed how the interaction between those processes modulates cell volume. Cells exposed to hypotonic medium swelled 1.5 times their basal volume. Swelling was followed by 41% RVD40 (extent of RVD after 40 min of maximum), whereas the concentration of extracellular ATP (ATPe) increased 10 times to a maximum value at 15 min. Exogenous apyrase (which removes di- and trinucleotides) did not alter RVD, whereas exogenous Na+-K+ ATPase (which converts ATP to ADP in the extracellular medium) enhanced RVD40 by 2.6 times, suggesting that hypotonic treatment alone produced a basal RVD, whereas extracellular ADP activated RVD to achieve complete volume regulation (i.e., RVD40≈100%). Under hypotonicity, addition of 2MetSADP (ADP analog) increased RVD to the same extent as exposure to Na+-K+ ATPase and the same analog did not stimulate RVD when co-incubated with MRS2211, a blocker of ADP receptor P2Y13. RT-PCR and Western blot analysis confirmed the presence of P2Y13. Cells exhibited significant ectoATPase activity, which according to RT-PCR analysis can be assigned to ENTPDase2. Both carbenoxolone, a blocker of conductive ATP release, and Brefeldin A, an inhibitor of exocytosis, were able to partially decrease ATPe accumulation, pointing to the presence of at least two mechanisms for ATP release. Thus, in Huh-7 cells, hypotonic treatment triggered the release of ATP. Conversion of ATPe to ADPe by ENTPDase 2 activity facilitates the accumulated ADPe to activate P2Y13 receptors, which mediate complete RVD.
dc.languageeng
dc.publisherAmerican Physiological Society
dc.relationinfo:eu-repo/semantics/altIdentifier/ark/http://ajpcell.physiology.org/content/304/10/C1013.long
dc.relationinfo:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1152/ajpcell.00254.2012
dc.rightshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectP2y13 Receptor
dc.subjectCell Volume Regulation
dc.subjectHuman Hepatoma
dc.subjectNucleotides
dc.subjectPurinergic Signaling
dc.titleOn the role of ATP release, ectoATPase activity, and extracellular ADP in the regulatory volume decrease of Huh-7 human hepatoma cells
dc.typeinfo:eu-repo/semantics/article
dc.typeinfo:ar-repo/semantics/artículo
dc.typeinfo:eu-repo/semantics/publishedVersion


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