A necessary connection: cholesterol and nicotinic receptors
- Autores
- Fabiani, Camila; Peñalva, Daniel Alejandro; Corradi, Jeremias; Antollini, Silvia Susana
- Año de publicación
- 2019
- Idioma
- inglés
- Tipo de recurso
- documento de conferencia
- Estado
- versión publicada
- Descripción
- It is known that the muscle nicotinic acetylcholine receptor (nAChR) is highly influenced by its lipid environment. It is present in high-density clusters in the muscle cell membrane where it localizes mainly in liquid-ordered (Lo) domains enriched in cholesterol and sphingolipids. Its transmembrane domain forms the ion channel pore and exhibits extensive contacts with the surrounding lipids. In this work we studied the close relationship between nAChR and cholesterol under different experimental conditions in order to enrich with, deplete of, redistribute between both hemilayers, and oxidate cholesterol molecules. These conditions were evaluated either in T. californica nAChR-rich membranes, in model membranes containing purified nAChR or in cells expressing nAChR. Cholesterol modifications were confirmed by lipid analysis using thin layer chromatography. Evaluation of a) membrane order perturbations, by Laurdan GP and fluorescence anisotropy, b) increase/decrease of Lo domains, by fluorescence microscopy, c) nAChR-Lo domains correlation, by detergent treatment and SDS-PAGE, and d) nAChR conformation and function, by fluorescence spectroscopy and electrophysiology showed that changes in the amount, distribution or oxidation of cholesterol impacts not only in the size, location and curvature-domain shape of Lo domains and in the nAChR preference for them, but also in nAChR functionality and nAChR structural conformation. A high correlation between the quantitative presence of cholesterol, its transmembrane and lateral asymmetry and nAChR conformation and functionality is postulated.
Fil: Fabiani, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Peñalva, Daniel Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Corradi, Jeremias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Antollini, Silvia Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
XLVIII Reunión Anual de la Sociedad Argentina de Biofísica
San Luis
Argentina
Sociedad Argentina de Biofísica - Materia
-
nicotinic acetylcholine receptor
cholesterol
ciclodextrin
cholesterol oxidase - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/161306
Ver los metadatos del registro completo
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A necessary connection: cholesterol and nicotinic receptorsFabiani, CamilaPeñalva, Daniel AlejandroCorradi, JeremiasAntollini, Silvia Susananicotinic acetylcholine receptorcholesterolciclodextrincholesterol oxidasehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1It is known that the muscle nicotinic acetylcholine receptor (nAChR) is highly influenced by its lipid environment. It is present in high-density clusters in the muscle cell membrane where it localizes mainly in liquid-ordered (Lo) domains enriched in cholesterol and sphingolipids. Its transmembrane domain forms the ion channel pore and exhibits extensive contacts with the surrounding lipids. In this work we studied the close relationship between nAChR and cholesterol under different experimental conditions in order to enrich with, deplete of, redistribute between both hemilayers, and oxidate cholesterol molecules. These conditions were evaluated either in T. californica nAChR-rich membranes, in model membranes containing purified nAChR or in cells expressing nAChR. Cholesterol modifications were confirmed by lipid analysis using thin layer chromatography. Evaluation of a) membrane order perturbations, by Laurdan GP and fluorescence anisotropy, b) increase/decrease of Lo domains, by fluorescence microscopy, c) nAChR-Lo domains correlation, by detergent treatment and SDS-PAGE, and d) nAChR conformation and function, by fluorescence spectroscopy and electrophysiology showed that changes in the amount, distribution or oxidation of cholesterol impacts not only in the size, location and curvature-domain shape of Lo domains and in the nAChR preference for them, but also in nAChR functionality and nAChR structural conformation. A high correlation between the quantitative presence of cholesterol, its transmembrane and lateral asymmetry and nAChR conformation and functionality is postulated.Fil: Fabiani, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Peñalva, Daniel Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Corradi, Jeremias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Antollini, Silvia Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaXLVIII Reunión Anual de la Sociedad Argentina de BiofísicaSan LuisArgentinaSociedad Argentina de BiofísicaSociedad Argentina de BiofísicaAndujar, Sebastian Antonio2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/161306A necessary connection: cholesterol and nicotinic receptors; XLVIII Reunión Anual de la Sociedad Argentina de Biofísica; San Luis; Argentina; 2019; 78-78978-987-27591-7-9CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/reuniones-cientificas/reunionsab-previas/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:48:48Zoai:ri.conicet.gov.ar:11336/161306instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:48:48.703CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
A necessary connection: cholesterol and nicotinic receptors |
title |
A necessary connection: cholesterol and nicotinic receptors |
spellingShingle |
A necessary connection: cholesterol and nicotinic receptors Fabiani, Camila nicotinic acetylcholine receptor cholesterol ciclodextrin cholesterol oxidase |
title_short |
A necessary connection: cholesterol and nicotinic receptors |
title_full |
A necessary connection: cholesterol and nicotinic receptors |
title_fullStr |
A necessary connection: cholesterol and nicotinic receptors |
title_full_unstemmed |
A necessary connection: cholesterol and nicotinic receptors |
title_sort |
A necessary connection: cholesterol and nicotinic receptors |
dc.creator.none.fl_str_mv |
Fabiani, Camila Peñalva, Daniel Alejandro Corradi, Jeremias Antollini, Silvia Susana |
author |
Fabiani, Camila |
author_facet |
Fabiani, Camila Peñalva, Daniel Alejandro Corradi, Jeremias Antollini, Silvia Susana |
author_role |
author |
author2 |
Peñalva, Daniel Alejandro Corradi, Jeremias Antollini, Silvia Susana |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
Andujar, Sebastian Antonio |
dc.subject.none.fl_str_mv |
nicotinic acetylcholine receptor cholesterol ciclodextrin cholesterol oxidase |
topic |
nicotinic acetylcholine receptor cholesterol ciclodextrin cholesterol oxidase |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
It is known that the muscle nicotinic acetylcholine receptor (nAChR) is highly influenced by its lipid environment. It is present in high-density clusters in the muscle cell membrane where it localizes mainly in liquid-ordered (Lo) domains enriched in cholesterol and sphingolipids. Its transmembrane domain forms the ion channel pore and exhibits extensive contacts with the surrounding lipids. In this work we studied the close relationship between nAChR and cholesterol under different experimental conditions in order to enrich with, deplete of, redistribute between both hemilayers, and oxidate cholesterol molecules. These conditions were evaluated either in T. californica nAChR-rich membranes, in model membranes containing purified nAChR or in cells expressing nAChR. Cholesterol modifications were confirmed by lipid analysis using thin layer chromatography. Evaluation of a) membrane order perturbations, by Laurdan GP and fluorescence anisotropy, b) increase/decrease of Lo domains, by fluorescence microscopy, c) nAChR-Lo domains correlation, by detergent treatment and SDS-PAGE, and d) nAChR conformation and function, by fluorescence spectroscopy and electrophysiology showed that changes in the amount, distribution or oxidation of cholesterol impacts not only in the size, location and curvature-domain shape of Lo domains and in the nAChR preference for them, but also in nAChR functionality and nAChR structural conformation. A high correlation between the quantitative presence of cholesterol, its transmembrane and lateral asymmetry and nAChR conformation and functionality is postulated. Fil: Fabiani, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina Fil: Peñalva, Daniel Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina Fil: Corradi, Jeremias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina Fil: Antollini, Silvia Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina XLVIII Reunión Anual de la Sociedad Argentina de Biofísica San Luis Argentina Sociedad Argentina de Biofísica |
description |
It is known that the muscle nicotinic acetylcholine receptor (nAChR) is highly influenced by its lipid environment. It is present in high-density clusters in the muscle cell membrane where it localizes mainly in liquid-ordered (Lo) domains enriched in cholesterol and sphingolipids. Its transmembrane domain forms the ion channel pore and exhibits extensive contacts with the surrounding lipids. In this work we studied the close relationship between nAChR and cholesterol under different experimental conditions in order to enrich with, deplete of, redistribute between both hemilayers, and oxidate cholesterol molecules. These conditions were evaluated either in T. californica nAChR-rich membranes, in model membranes containing purified nAChR or in cells expressing nAChR. Cholesterol modifications were confirmed by lipid analysis using thin layer chromatography. Evaluation of a) membrane order perturbations, by Laurdan GP and fluorescence anisotropy, b) increase/decrease of Lo domains, by fluorescence microscopy, c) nAChR-Lo domains correlation, by detergent treatment and SDS-PAGE, and d) nAChR conformation and function, by fluorescence spectroscopy and electrophysiology showed that changes in the amount, distribution or oxidation of cholesterol impacts not only in the size, location and curvature-domain shape of Lo domains and in the nAChR preference for them, but also in nAChR functionality and nAChR structural conformation. A high correlation between the quantitative presence of cholesterol, its transmembrane and lateral asymmetry and nAChR conformation and functionality is postulated. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/conferenceObject Reunión Book http://purl.org/coar/resource_type/c_5794 info:ar-repo/semantics/documentoDeConferencia |
status_str |
publishedVersion |
format |
conferenceObject |
dc.identifier.none.fl_str_mv |
http://hdl.handle.net/11336/161306 A necessary connection: cholesterol and nicotinic receptors; XLVIII Reunión Anual de la Sociedad Argentina de Biofísica; San Luis; Argentina; 2019; 78-78 978-987-27591-7-9 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/161306 |
identifier_str_mv |
A necessary connection: cholesterol and nicotinic receptors; XLVIII Reunión Anual de la Sociedad Argentina de Biofísica; San Luis; Argentina; 2019; 78-78 978-987-27591-7-9 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/reuniones-cientificas/reunionsab-previas/ |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
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application/pdf application/pdf application/pdf application/pdf application/pdf |
dc.coverage.none.fl_str_mv |
Nacional |
dc.publisher.none.fl_str_mv |
Sociedad Argentina de Biofísica |
publisher.none.fl_str_mv |
Sociedad Argentina de Biofísica |
dc.source.none.fl_str_mv |
reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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