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Regulatory volume decrease and intracellular Ca2+ in murine neuroblastoma cells studied with fluorescent probes

dc.creatorAltamirano, J.
dc.creatorBrodwick, M.S.
dc.creatorAlvarez-Leefmans, F.J.
dc.date.accessioned2017-06-29T04:22:15Z
dc.date.available2017-06-29T04:22:15Z
dc.date.issued1998es_ES
dc.identifier277es_ES
dc.identifier.issn0022-1295es_ES
dc.identifier.urihttps://doi.org/10.1085/jgp.112.2.145es_ES
dc.identifier.urihttp://repositorio.inprf.gob.mx/handle/123456789/4971
dc.language.isoenges_ES
dc.relation112 (2) 145-160 p.es_ES
dc.relationversión del editores_ES
dc.rightsacceso cerradoes_ES
dc.titleRegulatory volume decrease and intracellular Ca2+ in murine neuroblastoma cells studied with fluorescent probeses_ES
dc.typearticlees_ES
dc.contributor.affiliationDepartamento de Neurobiología, Instituto Mexicano de Psiquiatría, México, D.F. México.es_ES
dc.relation.jnabreviadoJ GEN PHYSIOLes_ES
dc.relation.journalThe Journal of general physiologyes_ES
dc.identifier.placeEstados Unidoses_ES
dc.date.published1998es_ES
dc.identifier.organizacionInstituto Mexicano de Psiquiatríaes_ES
dc.identifier.eissn1540-7748es_ES
dc.identifier.doi10.1085/jgp.112.2.145es_ES
dc.description.monthAgoes_ES
dc.description.abstractotrodiomaThe possible role of Ca2+ as a second messenger mediating regulatory volume decrease (RVD) in osmotically swollen cells was investigated in murine neural cell lines (N1E-115 and NG108-15) by means of novel microspectrofluorimetric techniques that allow simultaneous measurement of changes in cell water volume and [Ca2+]i in single cells loaded with fura-2. [Ca2+]i was measured ratiometrically, whereas the volume change was determined at the intracellular isosbestic wavelength (358 nm). Independent volume measurements were done using calcein, a fluorescent probe insensitive to intracellular ions. When challenged with _40% hyposmotic solutions, the cells expanded osmometrically and then underwent RVD. Concomitant with the volume response, there was a transient increase in [Ca2+]i, whose onset preceded RVD. For hyposmotic solutions (up to __40%), [Ca2+]i increased steeply with the reciprocal of the external osmotic pressure and with the cell volume. Chelation of external and internal Ca2+, with EGTA and 1,2-bis-(o_-aminophenoxy) ethane-N,N,N__,N__-tetraacetic acid (BAPTA), respectively, attenuated but did not prevent RVD. This Ca2+-independent RVD proceeded even when there was a concomitant decrease in [Ca2+]i below resting levels. Similar results were obtained in cells loaded with calcein. For cells not treated with BAPTA, restoration of external Ca2+ during the relaxation of RVD elicited by Ca2+-free hyposmotic solutions produced an increase in [Ca2+]i without affecting the rate or extent of the responses. RVD and the increase in [Ca2+]i were blocked or attenuated upon the second of two _40% hyposmotic challenges applied at an interval of 30–60 min. The inactivation persisted in Ca2+-free solutions. Hence, our simultaneous measurements of intracellular Ca2+ and volume in single neuroblastoma cells directly demonstrate that an increase in intracellular Ca2+ is not necessary for triggering RVD or its inactivation. The attenuation of RVD after Ca2+ chelation could occur through secondary effects or could indicate that Ca2+ is required for optimal RVD responseses_ES
dc.subject.kwAnimalses_ES
dc.subject.kwCalcium-Metabolismes_ES
dc.subject.kwCalcium-Pharmacologyes_ES
dc.subject.kwCell Size-Physiologyes_ES
dc.subject.kwChelating Agents-Pharmacologyes_ES
dc.subject.kwEgtazic Acid-Analogs & derivativeses_ES
dc.subject.kwEgtazic Acid-Pharmacologyes_ES
dc.subject.kwFluoresceins-Pharmacokineticses_ES
dc.subject.kwFluorescent Dyes-Pharmacokineticses_ES
dc.subject.kwFura-2-Pharmacokineticses_ES
dc.subject.kwGliomaes_ES
dc.subject.kwHybrid Cells-Drug effectses_ES
dc.subject.kwHybrid Cells-Metabolismes_ES
dc.subject.kwHypotonic Solutions-Pharmacologyes_ES
dc.subject.kwMicees_ES
dc.subject.kwNeuroblastomaes_ES
dc.subject.kwNeurons-Cytologyes_ES
dc.subject.kwNeurons-Drug effectses_ES
dc.subject.kwNeurons-Metabolismes_ES
dc.subject.kwOsmolar Concentrationes_ES
dc.subject.kwRatses_ES
dc.subject.kwTumor Cells, Cultured-Drug effectses_ES
dc.subject.kwTumor Cells,es_ES
dc.subject.kwCultured-Metabolismes_ES
dc.subject.kwChelating Agentses_ES
dc.subject.kwFluoresceinses_ES
dc.subject.kwFluorescent Dyeses_ES
dc.subject.kwHypotonic Solutionses_ES
dc.subject.kwEgtazic Acides_ES
dc.subject.kwCalciumes_ES
dc.subject.kwFura-2es_ES
dc.subject.kw1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acides_ES
dc.subject.kwFluorexones_ES


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