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

Autores
Kamerbeek, Constanza B.; Borroni, María Virginia; Pediconi, María F.; Sato, Satoshi B.; Kobayashi, Toshihide; Barrantes, Francisco José
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Fil: Kamerbeek, Constanza B. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Borroni, María Virginia. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Pediconi, María F. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Sato, Satoshi B. Lipid Biology Laboratory, RIKEN; Japón
Fil: Kobayashi, Toshihide. Lipid Biology Laboratory, RIKEN; Japón
Fil: Barrantes, Francisco José. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Neurobiología Molecular; Argentina
Fil: Barrantes Francisco José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Barrantes, Francisco José. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Abstract: 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 (α-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.
Fuente
Biophysical Journal Vol. 105, N° 7, 2013
Materia
MEDICINA
RECEPTORES
MEMBRANAS CELULARES
COLESTEROL
ANTICUERPOS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
Repositorio Institucional (UCA)
Institución
Pontificia Universidad Católica Argentina
OAI Identificador
oai:ucacris:123456789/8752

id RIUCA_ae67f14b7e3e5442ba9961b371ddd0dd
oai_identifier_str oai:ucacris:123456789/8752
network_acronym_str RIUCA
repository_id_str 2585
network_name_str Repositorio Institucional (UCA)
spelling Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domainsKamerbeek, Constanza B.Borroni, María VirginiaPediconi, María F.Sato, Satoshi B.Kobayashi, ToshihideBarrantes, Francisco JoséMEDICINARECEPTORESMEMBRANAS CELULARESCOLESTEROLANTICUERPOSFil: Kamerbeek, Constanza B. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Borroni, María Virginia. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Pediconi, María F. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Sato, Satoshi B. Lipid Biology Laboratory, RIKEN; JapónFil: Kobayashi, Toshihide. Lipid Biology Laboratory, RIKEN; JapónFil: Barrantes, Francisco José. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Neurobiología Molecular; ArgentinaFil: Barrantes Francisco José. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Barrantes, Francisco José. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaAbstract: 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 (α-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.Elsevier B.V.2013info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttps://repositorio.uca.edu.ar/handle/123456789/87520006-3495 (impreso)1542-0086 (online)10.1016/j.bpj.2013.08.03924094401Kamerbeek CB, Borroni V, Pediconi MF, Sato SB, Kobayashi T, Barrantes FJ. Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains. Biophys J. 2013;105(7):1601–1611. doi:10.1016/j.bpj.2013.08.039. Disponible en: https://repositorio.uca.edu.ar/handle/123456789/8752Biophysical Journal Vol. 105, N° 7, 2013reponame:Repositorio Institucional (UCA)instname:Pontificia Universidad Católica Argentinaenginfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/4.0/2025-07-03T10:56:54Zoai:ucacris:123456789/8752instacron:UCAInstitucionalhttps://repositorio.uca.edu.ar/Universidad privadaNo correspondehttps://repositorio.uca.edu.ar/oaiclaudia_fernandez@uca.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:25852025-07-03 10:56:55.188Repositorio Institucional (UCA) - Pontificia Universidad Católica Argentinafalse
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 B.
MEDICINA
RECEPTORES
MEMBRANAS CELULARES
COLESTEROL
ANTICUERPOS
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 B.
Borroni, María Virginia
Pediconi, María F.
Sato, Satoshi B.
Kobayashi, Toshihide
Barrantes, Francisco José
author Kamerbeek, Constanza B.
author_facet Kamerbeek, Constanza B.
Borroni, María Virginia
Pediconi, María F.
Sato, Satoshi B.
Kobayashi, Toshihide
Barrantes, Francisco José
author_role author
author2 Borroni, María Virginia
Pediconi, María F.
Sato, Satoshi B.
Kobayashi, Toshihide
Barrantes, Francisco José
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv MEDICINA
RECEPTORES
MEMBRANAS CELULARES
COLESTEROL
ANTICUERPOS
topic MEDICINA
RECEPTORES
MEMBRANAS CELULARES
COLESTEROL
ANTICUERPOS
dc.description.none.fl_txt_mv Fil: Kamerbeek, Constanza B. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Borroni, María Virginia. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Pediconi, María F. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Sato, Satoshi B. Lipid Biology Laboratory, RIKEN; Japón
Fil: Kobayashi, Toshihide. Lipid Biology Laboratory, RIKEN; Japón
Fil: Barrantes, Francisco José. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas. Laboratorio de Neurobiología Molecular; Argentina
Fil: Barrantes Francisco José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Barrantes, Francisco José. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Abstract: 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 (α-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.
description Fil: Kamerbeek, Constanza B. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
publishDate 2013
dc.date.none.fl_str_mv 2013
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 https://repositorio.uca.edu.ar/handle/123456789/8752
0006-3495 (impreso)
1542-0086 (online)
10.1016/j.bpj.2013.08.039
24094401
Kamerbeek CB, Borroni V, Pediconi MF, Sato SB, Kobayashi T, Barrantes FJ. Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains. Biophys J. 2013;105(7):1601–1611. doi:10.1016/j.bpj.2013.08.039. Disponible en: https://repositorio.uca.edu.ar/handle/123456789/8752
url https://repositorio.uca.edu.ar/handle/123456789/8752
identifier_str_mv 0006-3495 (impreso)
1542-0086 (online)
10.1016/j.bpj.2013.08.039
24094401
Kamerbeek CB, Borroni V, Pediconi MF, Sato SB, Kobayashi T, Barrantes FJ. Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains. Biophys J. 2013;105(7):1601–1611. doi:10.1016/j.bpj.2013.08.039. Disponible en: https://repositorio.uca.edu.ar/handle/123456789/8752
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/4.0/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/4.0/
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier B.V.
publisher.none.fl_str_mv Elsevier B.V.
dc.source.none.fl_str_mv Biophysical Journal Vol. 105, N° 7, 2013
reponame:Repositorio Institucional (UCA)
instname:Pontificia Universidad Católica Argentina
reponame_str Repositorio Institucional (UCA)
collection Repositorio Institucional (UCA)
instname_str Pontificia Universidad Católica Argentina
repository.name.fl_str_mv Repositorio Institucional (UCA) - Pontificia Universidad Católica Argentina
repository.mail.fl_str_mv claudia_fernandez@uca.edu.ar
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score 13.070432