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dc.contributor.author Fraire, Juan Carlos
dc.contributor.author Masseroni, Maria Luján
dc.contributor.author Jausoro, Ignacio
dc.contributor.author Perassi, Eduardo Marcelo
dc.contributor.author Diaz Añel, Alberto Marcelo
dc.contributor.author Coronado, Eduardo A.
dc.date.available 2017-12-27T16:08:24Z
dc.date.issued 2014-09
dc.identifier.citation Coronado, Eduardo A.; Perassi, Eduardo Marcelo; Jausoro, Ignacio; Masseroni, Maria Luján; Fraire, Juan Carlos; Diaz Añel, Alberto Marcelo; et al.; Identification, Localization, and Quantification of Neuronal Cell Membrane Receptors with Plasmonic Probes: Role of Protein Kinase D1 in Their Distribution; American Chemical Society; ACS Nano; 8; 9; 9-2014; 8942-8958
dc.identifier.issn 1936-0851
dc.identifier.uri http://hdl.handle.net/11336/31650
dc.description.abstract Detecting, imaging, and being able to localize the distribution ofseveral cell membrane receptors on a single neuron are very important topics inneuroscience research. In the present work, the distribution of metabotropicglutamate receptor 1a (mGluR1a) density on neuron cells on subcellular lengthscales is determined by evaluating the role played by protein kinase D1 (PKD1) inthe trafficking of membrane proteins, comparing the distribution of mGluR1a inexperiments performed in endogenous PKD1 expression with those in the presenceof kinase-inactive protein kinase D1 (PKD1-kd). The localization, distribution, anddensity of cell surface mGluR1a were evaluated using 90 nm diameter Aunanoparticle (NP) probes specifically functionalized with a high-affinity and multivalent labeling function, which allows not only imaging NPs wherethis receptor is present but also quantifying by optical means the NP density. This is so because the NP generates a density (F)-dependent SERS responsethat facilitated a spatial mapping of the mGluR1a density distribution on subcellular length scales (dendrites and axons) in an optical microscope.ThemeasuredFvalues were found to be significantly higher on dendrites than on axons for endogenous PKD1, while an increase ofFon axons was observedwhen PKD1 is altered. The spatial distribution of the NP immunolabels through scanning electron microscopy (SEM) confirmed the results obtained byfluorescence bright-field analysis and dark-field spectroscopy and provided additional structural details. In addition, it is shown using electrodynamicsimulations that SERS spectroscopy could be a very sensitive tool for the spatial mapping of cell membrane receptors on subcellular length scales, asSERSsignals are almost linearly dependent on NP density and therefore give indirect information on the distribution of cell membrane proteins. This result isimportant since the calibration of theF-dependent near-field enhancement of the Au immunolabels through correlation of SERS and SEM paves the waytoward quantitative immunolabeling studies of cell membrane proteins involved in neuron polarity. From the molecular biology point of view, this studyshows that in cultured hippocampal pyramidal cells mGluR1a is predominantly transported to dendrites and excluded from axons. Expression of kinase-inactive protein kinase D1 (PKD1-kd) dramatically and selectively alters the intracellular trafficking and membrane delivery of mGluR1a-containing vesicles.
dc.format application/pdf
dc.language.iso eng
dc.publisher American Chemical Society
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject plasmonics
dc.subject nanoparticles
dc.subject bioconjugation
dc.subject immunolabels
dc.subject sensing
dc.subject neurons
dc.subject PKD1
dc.subject sorting
dc.subject neurotransmitter receptors
dc.subject.classification Otras Ciencias Biológicas
dc.subject.classification Ciencias Biológicas
dc.subject.classification CIENCIAS NATURALES Y EXACTAS
dc.subject.classification Astronomía
dc.subject.classification Ciencias Físicas
dc.subject.classification CIENCIAS NATURALES Y EXACTAS
dc.title Identification, Localization, and Quantification of Neuronal Cell Membrane Receptors with Plasmonic Probes: Role of Protein Kinase D1 in Their Distribution
dc.type info:eu-repo/semantics/article
dc.type info:ar-repo/semantics/artículo
dc.type info:eu-repo/semantics/publishedVersion
dc.date.updated 2017-12-26T20:36:46Z
dc.journal.volume 8
dc.journal.number 9
dc.journal.pagination 8942-8958
dc.journal.pais Estados Unidos
dc.journal.ciudad Washington
dc.description.fil Fil: Fraire, Juan Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina
dc.description.fil Fil: Masseroni, Maria Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
dc.description.fil Fil: Jausoro, Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
dc.description.fil Fil: Perassi, Eduardo Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina
dc.description.fil Fil: Diaz Añel, Alberto Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
dc.description.fil Fil: Coronado, Eduardo A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina
dc.journal.title ACS Nano
dc.relation.alternativeid info:eu-repo/semantics/altIdentifier/url/http://pubs.acs.org/doi/10.1021/nn501575c
dc.relation.alternativeid info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1021/nn501575c
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info:eu-repo/semantics/restrictedAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)