Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains

Autores
Kamerbeek, Constanza Belén; Borroni, Maria Virginia; Pediconi, Maria Filomena; Sato, Satoshi B.; Kobayashi, Toshihide; Barrantes, Francisco Jose
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The distribution of nicotinic acetylcholine receptor (AChR) clusters at the cell membrane was studied in CHO-K1/A5 cells using fluorescence microscopy. Di-4-ANEPPDHQ, a fluorescent probe that differentiates between liquid-ordered (Lo) and liquid-disordered (Ld) phases in model membranes, was used in combination with monoclonal anti-AChR antibody labeling of live cells, which induces AChR clustering. The so-called generalized polarization (GP) of di-4-ANEPPDHQ was measured in regions of the cell-surface membrane associated with or devoid of antibody-induced AChR clusters, respectively. AChR clusters were almost equally distributed between Lo and Ld domains, independently of receptor surface levels and agonist (carbamoylcholine and nicotine) or antagonist (á-bungarotoxin) binding. Cholesterol depletion diminished the cell membrane mean di-4-ANEPPDHQ GP and the number of AChR clusters associated with Ld membrane domains increased concomitantly. Depolymerization of the filamentous actin cytoskeleton by Latrunculin A had the opposite effect, with more AChR clusters associated with Lo domains. AChR internalized via small vesicles having lower GP and lower cholesterol content than the surface membrane. Upon cholesterol depletion, only 12% of the AChR-containing vesicles costained with the fluorescent cholesterol analog fPEG-cholesterol, i.e., AChR endocytosis was essentially dissociated from that of cholesterol. In conclusion, the distribution of AChR submicron-sized clusters at the cell membrane appears to be regulated by cholesterol content and cytoskeleton integrity
Fil: Kamerbeek, Constanza Belén. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); Argentina
Fil: Borroni, Maria Virginia. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); Argentina
Fil: Pediconi, Maria Filomena. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); Argentina
Fil: Sato, Satoshi B.. Lipid Biology Laboratory; Japón
Fil: Kobayashi, Toshihide. Lipid Biology Laboratory; Japón
Fil: Barrantes, Francisco Jose. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Investigaciones Biomedicas; Argentina
Materia
Nicotinic Receptor
Lipid Domains
Di-4-Aneppdhq
Generalized Polarization
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/5260
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/5260
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domainsKamerbeek, Constanza BelénBorroni, Maria VirginiaPediconi, Maria FilomenaSato, Satoshi B.Kobayashi, ToshihideBarrantes, Francisco JoseNicotinic ReceptorLipid DomainsDi-4-AneppdhqGeneralized Polarizationhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The distribution of nicotinic acetylcholine receptor (AChR) clusters at the cell membrane was studied in CHO-K1/A5 cells using fluorescence microscopy. Di-4-ANEPPDHQ, a fluorescent probe that differentiates between liquid-ordered (Lo) and liquid-disordered (Ld) phases in model membranes, was used in combination with monoclonal anti-AChR antibody labeling of live cells, which induces AChR clustering. The so-called generalized polarization (GP) of di-4-ANEPPDHQ was measured in regions of the cell-surface membrane associated with or devoid of antibody-induced AChR clusters, respectively. AChR clusters were almost equally distributed between Lo and Ld domains, independently of receptor surface levels and agonist (carbamoylcholine and nicotine) or antagonist (á-bungarotoxin) binding. Cholesterol depletion diminished the cell membrane mean di-4-ANEPPDHQ GP and the number of AChR clusters associated with Ld membrane domains increased concomitantly. Depolymerization of the filamentous actin cytoskeleton by Latrunculin A had the opposite effect, with more AChR clusters associated with Lo domains. AChR internalized via small vesicles having lower GP and lower cholesterol content than the surface membrane. Upon cholesterol depletion, only 12% of the AChR-containing vesicles costained with the fluorescent cholesterol analog fPEG-cholesterol, i.e., AChR endocytosis was essentially dissociated from that of cholesterol. In conclusion, the distribution of AChR submicron-sized clusters at the cell membrane appears to be regulated by cholesterol content and cytoskeleton integrityFil: Kamerbeek, Constanza Belén. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); ArgentinaFil: Borroni, Maria Virginia. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); ArgentinaFil: Pediconi, Maria Filomena. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); ArgentinaFil: Sato, Satoshi B.. Lipid Biology Laboratory; JapónFil: Kobayashi, Toshihide. Lipid Biology Laboratory; JapónFil: Barrantes, Francisco Jose. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Investigaciones Biomedicas; ArgentinaElsevier2013-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/5260Kamerbeek, Constanza Belén; Borroni, Maria Virginia; Pediconi, Maria Filomena; Sato, Satoshi B.; Kobayashi, Toshihide; et al.; Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains; Elsevier; Biophysical Journal; 105; 7; 10-2013; 1601-16110006-3495enginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0006349513009867info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpj.2013.08.039info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:40:37Zoai:ri.conicet.gov.ar:11336/5260instacron: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 10:40:37.517CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
title Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
spellingShingle Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
Kamerbeek, Constanza Belén
Nicotinic Receptor
Lipid Domains
Di-4-Aneppdhq
Generalized Polarization
title_short Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
title_full Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
title_fullStr Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
title_full_unstemmed Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
title_sort Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
dc.creator.none.fl_str_mv Kamerbeek, Constanza Belén
Borroni, Maria Virginia
Pediconi, Maria Filomena
Sato, Satoshi B.
Kobayashi, Toshihide
Barrantes, Francisco Jose
author Kamerbeek, Constanza Belén
author_facet Kamerbeek, Constanza Belén
Borroni, Maria Virginia
Pediconi, Maria Filomena
Sato, Satoshi B.
Kobayashi, Toshihide
Barrantes, Francisco Jose
author_role author
author2 Borroni, Maria Virginia
Pediconi, Maria Filomena
Sato, Satoshi B.
Kobayashi, Toshihide
Barrantes, Francisco Jose
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Nicotinic Receptor
Lipid Domains
Di-4-Aneppdhq
Generalized Polarization
topic Nicotinic Receptor
Lipid Domains
Di-4-Aneppdhq
Generalized Polarization
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The distribution of nicotinic acetylcholine receptor (AChR) clusters at the cell membrane was studied in CHO-K1/A5 cells using fluorescence microscopy. Di-4-ANEPPDHQ, a fluorescent probe that differentiates between liquid-ordered (Lo) and liquid-disordered (Ld) phases in model membranes, was used in combination with monoclonal anti-AChR antibody labeling of live cells, which induces AChR clustering. The so-called generalized polarization (GP) of di-4-ANEPPDHQ was measured in regions of the cell-surface membrane associated with or devoid of antibody-induced AChR clusters, respectively. AChR clusters were almost equally distributed between Lo and Ld domains, independently of receptor surface levels and agonist (carbamoylcholine and nicotine) or antagonist (á-bungarotoxin) binding. Cholesterol depletion diminished the cell membrane mean di-4-ANEPPDHQ GP and the number of AChR clusters associated with Ld membrane domains increased concomitantly. Depolymerization of the filamentous actin cytoskeleton by Latrunculin A had the opposite effect, with more AChR clusters associated with Lo domains. AChR internalized via small vesicles having lower GP and lower cholesterol content than the surface membrane. Upon cholesterol depletion, only 12% of the AChR-containing vesicles costained with the fluorescent cholesterol analog fPEG-cholesterol, i.e., AChR endocytosis was essentially dissociated from that of cholesterol. In conclusion, the distribution of AChR submicron-sized clusters at the cell membrane appears to be regulated by cholesterol content and cytoskeleton integrity
Fil: Kamerbeek, Constanza Belén. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); Argentina
Fil: Borroni, Maria Virginia. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); Argentina
Fil: Pediconi, Maria Filomena. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); Argentina
Fil: Sato, Satoshi B.. Lipid Biology Laboratory; Japón
Fil: Kobayashi, Toshihide. Lipid Biology Laboratory; Japón
Fil: Barrantes, Francisco Jose. Consejo Nacional de Investigaciones Cienti­ficas y Tecnicas. Centro Cientifico Tecnologico Bahi­a Blanca. Instituto de Investigaciones Bioquimicas Bahi­a Blanca (i); Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Investigaciones Biomedicas; Argentina
description The distribution of nicotinic acetylcholine receptor (AChR) clusters at the cell membrane was studied in CHO-K1/A5 cells using fluorescence microscopy. Di-4-ANEPPDHQ, a fluorescent probe that differentiates between liquid-ordered (Lo) and liquid-disordered (Ld) phases in model membranes, was used in combination with monoclonal anti-AChR antibody labeling of live cells, which induces AChR clustering. The so-called generalized polarization (GP) of di-4-ANEPPDHQ was measured in regions of the cell-surface membrane associated with or devoid of antibody-induced AChR clusters, respectively. AChR clusters were almost equally distributed between Lo and Ld domains, independently of receptor surface levels and agonist (carbamoylcholine and nicotine) or antagonist (á-bungarotoxin) binding. Cholesterol depletion diminished the cell membrane mean di-4-ANEPPDHQ GP and the number of AChR clusters associated with Ld membrane domains increased concomitantly. Depolymerization of the filamentous actin cytoskeleton by Latrunculin A had the opposite effect, with more AChR clusters associated with Lo domains. AChR internalized via small vesicles having lower GP and lower cholesterol content than the surface membrane. Upon cholesterol depletion, only 12% of the AChR-containing vesicles costained with the fluorescent cholesterol analog fPEG-cholesterol, i.e., AChR endocytosis was essentially dissociated from that of cholesterol. In conclusion, the distribution of AChR submicron-sized clusters at the cell membrane appears to be regulated by cholesterol content and cytoskeleton integrity
publishDate 2013
dc.date.none.fl_str_mv 2013-10
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/5260
Kamerbeek, Constanza Belén; Borroni, Maria Virginia; Pediconi, Maria Filomena; Sato, Satoshi B.; Kobayashi, Toshihide; et al.; Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains; Elsevier; Biophysical Journal; 105; 7; 10-2013; 1601-1611
0006-3495
url http://hdl.handle.net/11336/5260
identifier_str_mv Kamerbeek, Constanza Belén; Borroni, Maria Virginia; Pediconi, Maria Filomena; Sato, Satoshi B.; Kobayashi, Toshihide; et al.; Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains; Elsevier; Biophysical Journal; 105; 7; 10-2013; 1601-1611
0006-3495
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0006349513009867
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpj.2013.08.039
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
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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