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Immunolocalization of the Na+–K+–2Cl_ cotransporter in peripheral nervous tissue of vertebrates

dc.creatorAlvarez-Leefmans, F.J.
dc.creatorLeón-Olea, M.
dc.creatorMendoza-Sotelo, J.
dc.creatorAlvarez, F.J.
dc.creatorAntón, B.
dc.creatorGarduño, R.
dc.date.accessioned2017-06-29T04:26:10Z
dc.date.available2017-06-29T04:26:10Z
dc.date.issued2001es_ES
dc.identifier342es_ES
dc.identifier.issn0306-4522es_ES
dc.identifier.urihttp://repositorio.inprf.gob.mx/handle/123456789/5033
dc.identifier.urihttps://doi.org/10.1016/S0306-4522(01)00091-4es_ES
dc.language.isoenges_ES
dc.relation104 (2) 569–582 p.es_ES
dc.relationversión del editores_ES
dc.rightsacceso cerradoes_ES
dc.titleImmunolocalization of the Na+–K+–2Cl_ cotransporter in peripheral nervous tissue of vertebrateses_ES
dc.typearticlees_ES
dc.contributor.affiliationDepartment of Pharmacobiology, Centro de Investigación y de Estudios Avanzados, Instituto Politécnico Nacional, Apartado Postal 14-740, México D.F. 07000,es_ES
dc.relation.jnabreviadoNEUROSCIes_ES
dc.relation.journalNeurosciencees_ES
dc.identifier.placeUnited Stateses_ES
dc.date.published2001es_ES
dc.identifier.organizacionInstituto Nacional de Psiquiatría Ramón de la Fuente Muñizes_ES
dc.description.monthMayes_ES
dc.description.abstractotrodiomaEfflux of Cl_ through GABAA-gated anion channels depolarizes the cell bodies and intraspinal terminals of sensory neurons, and contributes to the generation of presynaptic inhibition in the spinal cord. Active accumulation of Cl_ inside sensory neurons occurs through an Na+–K+–2Cl_ cotransport system that generates and maintains the electrochemical gradient for this outward Cl_ current. We studied the immunolocalization of the Na+–K+–2Cl_ cotransporter protein using a monoclonal antibody (T4) against a conserved epitope in the C-terminus of the molecule. Western blots of frog, rat and cat dorsal root ganglion membranes revealed a single band of cotransporter immunoreactivity at _160 kDa, consistent with the molecular mass of the glycosylated protein. Deglycosylation with N-glycosidase F reduced the molecular mass to _135 kDa, in agreement with the size of the core polypeptide. Indirect immunofluorescence revealed strong cotransporter immunoreactivity in all types of dorsal root ganglion cell bodies in frog, rat and cat. The subcellular distribution of cotransporter immunoreactivity was different amongst species. Membrane labeling was more apparent in frog and rat dorsal root ganglion cell bodies than in cat. In contrast, cytoplasmic labeling was intense in cat and weak in frog, being intermediate in the rat. Cotransporter immunoreactivity also occurred in satellite cells, particularly in rat and cat dorsal root ganglia. The membrane region and axoplasm of sensory fibers were heavily labeled in cat and rat and less in frog. Three-dimensional reconstruction of confocal optical sections and dual immunolocalization with S-100 protein showed that the cotransporter immunoreactivity was prominently expressed in the nodal and paranodal regions of the Schwann cells. Ultrastructural immunolocalization confirmed the presence of immunoreactivity on the membranes of the axon and the Schwan cell in both the nodal region and the paranode. Treatment with sodium dodecylsulfate and _-mercaptoethanol also uncovered intense cotransporter immunoreactivity in Schmidt–Lanterman incisures at the light microscopic level. The localization of the Na+–K+–2Cl_ cotransporter protein is consistent with its function as a Cl_-accumulating mechanism in sensory neurons. Its distinctive presence in Schwann cells suggests that it could also be involved in K+ uptake from the extracellular space, particularly in the paranodal region of myelinated axons, thereby regulating the extracellular ionic environment and the excitability of axons.es_ES
dc.subject.koNa+–K+–Cl_ cotransportes_ES
dc.subject.koDRG cellses_ES
dc.subject.koSchwann celles_ES
dc.subject.koPresynaptic inhibitiones_ES
dc.subject.koPotassium bufferinges_ES
dc.subject.koIntracellular Cl_es_ES


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