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
- Institución
- Pontificia Universidad Católica Argentina
- OAI Identificador
- oai:ucacris:123456789/8752
Ver los metadatos del registro completo
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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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1836638347778326528 |
score |
13.070432 |