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
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/161306
Acceder
id CONICETDig_fbfe78d744c44fc5bbdf75bf3e97a28f
oai_identifier_str oai:ri.conicet.gov.ar:11336/161306
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling 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 Técnicas2025-09-29T09:42:14Zoai: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:34982025-09-29 09:42:14.717CONICET 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/
dc.format.none.fl_str_mv 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
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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