Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane

Autores
Borroni, Maria Virginia; Baier, Carlos Javier; Lang, T; Bonini, Ida Clara; White, M. M; Garbus, Ingrid; Barrantes, Francisco Jose
Año de publicación
2007
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Novel effects of cholesterol (Chol) on nicotinic acetylcholine receptor (AChR) cell-surface stability, internalization and function are reported. AChRs are shown to occur in the form of submicron-sized (240–280 nm) domains that remain stable at the cell-surface membrane of CHO-K1/A5 cells over a period of hours. Acute (30 min, 37°C) exposure to methyl-β-cyclodextrin (CDx), commonly used as a diagnostic tool of endocytic mechanisms, is shown here to enhance AChR internalization kinetics in the receptor-expressing clonal cell line. This treatment drastically reduced (∼50%) the number of receptor domains by accelerating the rate of endocytosis (t1/2 decreased from 1.5–0.5 h). In addition, Chol depletion produced ion channel gain-of-function of the remaining cell-surface AChR, whereas Chol enrichment had the opposite effect. Fluorescence measurements under conditions of direct excitation of the probe Laurdan and of Förster-type resonance energy transfer (FRET) using the intrinsic protein fluorescence as donor both indicated an increase in membrane fluidity in the bulk membrane and in the immediate environment of the AChR protein upon Chol depletion. Homeostatic control of Chol content at the plasmalemma may thus modulate cell-surface organization and stability of receptor domains, and fine tune receptor channel function to temporarily compensate for acute AChR loss from the cell surface.
Fil: Borroni, Maria Virginia. 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. Unesco; Argentina
Fil: Baier, Carlos Javier. 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: Lang, T. Max-Planck-Institut für biophysikalische Chemie; Alemania. Institut Max Planck fuer Bioanorganische Chemie; Alemania
Fil: Bonini, Ida Clara. 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: White, M. M. Drexel University; Estados Unidos
Fil: Garbus, Ingrid. 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: Barrantes, Francisco Jose. 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. Unesco; Argentina
Materia
CHOLESTEROL
CYCLODEXTRINS
ENDOCYTOSIS
FLUORESCENCE MICROSCOPY
LIPID DOMAINS
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:11336/150904
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/150904
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membraneBorroni, Maria VirginiaBaier, Carlos JavierLang, TBonini, Ida ClaraWhite, M. MGarbus, IngridBarrantes, Francisco JoseCHOLESTEROLCYCLODEXTRINSENDOCYTOSISFLUORESCENCE MICROSCOPYLIPID DOMAINShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Novel effects of cholesterol (Chol) on nicotinic acetylcholine receptor (AChR) cell-surface stability, internalization and function are reported. AChRs are shown to occur in the form of submicron-sized (240–280 nm) domains that remain stable at the cell-surface membrane of CHO-K1/A5 cells over a period of hours. Acute (30 min, 37°C) exposure to methyl-β-cyclodextrin (CDx), commonly used as a diagnostic tool of endocytic mechanisms, is shown here to enhance AChR internalization kinetics in the receptor-expressing clonal cell line. This treatment drastically reduced (∼50%) the number of receptor domains by accelerating the rate of endocytosis (t1/2 decreased from 1.5–0.5 h). In addition, Chol depletion produced ion channel gain-of-function of the remaining cell-surface AChR, whereas Chol enrichment had the opposite effect. Fluorescence measurements under conditions of direct excitation of the probe Laurdan and of Förster-type resonance energy transfer (FRET) using the intrinsic protein fluorescence as donor both indicated an increase in membrane fluidity in the bulk membrane and in the immediate environment of the AChR protein upon Chol depletion. Homeostatic control of Chol content at the plasmalemma may thus modulate cell-surface organization and stability of receptor domains, and fine tune receptor channel function to temporarily compensate for acute AChR loss from the cell surface.Fil: Borroni, Maria Virginia. 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. Unesco; ArgentinaFil: Baier, Carlos Javier. 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: Lang, T. Max-Planck-Institut für biophysikalische Chemie; Alemania. Institut Max Planck fuer Bioanorganische Chemie; AlemaniaFil: Bonini, Ida Clara. 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: White, M. M. Drexel University; Estados UnidosFil: Garbus, Ingrid. 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: Barrantes, Francisco Jose. 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. Unesco; ArgentinaTaylor & Francis Ltd2007-01-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/150904Borroni, Maria Virginia; Baier, Carlos Javier; Lang, T; Bonini, Ida Clara; White, M. M; et al.; Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane; Taylor & Francis Ltd; Molecular Membrane Biology; 24; 1; 9-1-2007; 1-150968-7688CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.tandfonline.com/doi/full/10.1080/09687860600903387info:eu-repo/semantics/altIdentifier/doi/10.1080/09687860600903387info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:17:08Zoai:ri.conicet.gov.ar:11336/150904instacron: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:34982025-10-15 15:17:08.952CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane
title Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane
spellingShingle Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane
Borroni, Maria Virginia
CHOLESTEROL
CYCLODEXTRINS
ENDOCYTOSIS
FLUORESCENCE MICROSCOPY
LIPID DOMAINS
title_short Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane
title_full Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane
title_fullStr Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane
title_full_unstemmed Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane
title_sort Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane
dc.creator.none.fl_str_mv Borroni, Maria Virginia
Baier, Carlos Javier
Lang, T
Bonini, Ida Clara
White, M. M
Garbus, Ingrid
Barrantes, Francisco Jose
author Borroni, Maria Virginia
author_facet Borroni, Maria Virginia
Baier, Carlos Javier
Lang, T
Bonini, Ida Clara
White, M. M
Garbus, Ingrid
Barrantes, Francisco Jose
author_role author
author2 Baier, Carlos Javier
Lang, T
Bonini, Ida Clara
White, M. M
Garbus, Ingrid
Barrantes, Francisco Jose
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv CHOLESTEROL
CYCLODEXTRINS
ENDOCYTOSIS
FLUORESCENCE MICROSCOPY
LIPID DOMAINS
topic CHOLESTEROL
CYCLODEXTRINS
ENDOCYTOSIS
FLUORESCENCE MICROSCOPY
LIPID DOMAINS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Novel effects of cholesterol (Chol) on nicotinic acetylcholine receptor (AChR) cell-surface stability, internalization and function are reported. AChRs are shown to occur in the form of submicron-sized (240–280 nm) domains that remain stable at the cell-surface membrane of CHO-K1/A5 cells over a period of hours. Acute (30 min, 37°C) exposure to methyl-β-cyclodextrin (CDx), commonly used as a diagnostic tool of endocytic mechanisms, is shown here to enhance AChR internalization kinetics in the receptor-expressing clonal cell line. This treatment drastically reduced (∼50%) the number of receptor domains by accelerating the rate of endocytosis (t1/2 decreased from 1.5–0.5 h). In addition, Chol depletion produced ion channel gain-of-function of the remaining cell-surface AChR, whereas Chol enrichment had the opposite effect. Fluorescence measurements under conditions of direct excitation of the probe Laurdan and of Förster-type resonance energy transfer (FRET) using the intrinsic protein fluorescence as donor both indicated an increase in membrane fluidity in the bulk membrane and in the immediate environment of the AChR protein upon Chol depletion. Homeostatic control of Chol content at the plasmalemma may thus modulate cell-surface organization and stability of receptor domains, and fine tune receptor channel function to temporarily compensate for acute AChR loss from the cell surface.
Fil: Borroni, Maria Virginia. 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. Unesco; Argentina
Fil: Baier, Carlos Javier. 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: Lang, T. Max-Planck-Institut für biophysikalische Chemie; Alemania. Institut Max Planck fuer Bioanorganische Chemie; Alemania
Fil: Bonini, Ida Clara. 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: White, M. M. Drexel University; Estados Unidos
Fil: Garbus, Ingrid. 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: Barrantes, Francisco Jose. 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. Unesco; Argentina
description Novel effects of cholesterol (Chol) on nicotinic acetylcholine receptor (AChR) cell-surface stability, internalization and function are reported. AChRs are shown to occur in the form of submicron-sized (240–280 nm) domains that remain stable at the cell-surface membrane of CHO-K1/A5 cells over a period of hours. Acute (30 min, 37°C) exposure to methyl-β-cyclodextrin (CDx), commonly used as a diagnostic tool of endocytic mechanisms, is shown here to enhance AChR internalization kinetics in the receptor-expressing clonal cell line. This treatment drastically reduced (∼50%) the number of receptor domains by accelerating the rate of endocytosis (t1/2 decreased from 1.5–0.5 h). In addition, Chol depletion produced ion channel gain-of-function of the remaining cell-surface AChR, whereas Chol enrichment had the opposite effect. Fluorescence measurements under conditions of direct excitation of the probe Laurdan and of Förster-type resonance energy transfer (FRET) using the intrinsic protein fluorescence as donor both indicated an increase in membrane fluidity in the bulk membrane and in the immediate environment of the AChR protein upon Chol depletion. Homeostatic control of Chol content at the plasmalemma may thus modulate cell-surface organization and stability of receptor domains, and fine tune receptor channel function to temporarily compensate for acute AChR loss from the cell surface.
publishDate 2007
dc.date.none.fl_str_mv 2007-01-09
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/11336/150904
Borroni, Maria Virginia; Baier, Carlos Javier; Lang, T; Bonini, Ida Clara; White, M. M; et al.; Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane; Taylor & Francis Ltd; Molecular Membrane Biology; 24; 1; 9-1-2007; 1-15
0968-7688
CONICET Digital
CONICET
url http://hdl.handle.net/11336/150904
identifier_str_mv Borroni, Maria Virginia; Baier, Carlos Javier; Lang, T; Bonini, Ida Clara; White, M. M; et al.; Cholesterol depletion activates rapid internalization of submicron-sized acetylcholine receptor domains at the cell membrane; Taylor & Francis Ltd; Molecular Membrane Biology; 24; 1; 9-1-2007; 1-15
0968-7688
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://www.tandfonline.com/doi/full/10.1080/09687860600903387
info:eu-repo/semantics/altIdentifier/doi/10.1080/09687860600903387
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
application/pdf
dc.publisher.none.fl_str_mv Taylor & Francis Ltd
publisher.none.fl_str_mv Taylor & Francis Ltd
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 13.22299

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