Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells

Autores
Borroni, Maria Virginia; Barrantes, Francisco Jose
Año de publicación
2010
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Stability of the nicotinic acetylcholine receptor (AChR) at the cell surface is critical to the correct functioning of the cholinergic synapse. Cholesterol (Chol) is an essential lipid that modulates AChR levels at the plasmalemma and ion translocation. We have studied the endocytosis of AChR in CHO-K1/A5 cells, a Chinese hamster ovary (CHO) cell line heterologously expressing murine muscle adult-type receptor under different Chol membrane content. Contrary to the norm, endocytosis of cell-surface AChR is accelerated by membrane Chol depletion via a hitherto unknown mechanism. This acceleration is no longer operative when membrane Chol levels are restored. We explored the possible mechanism involved in receptor loss in Choldepleted cells (Chol-). Under such conditions the AChR is internalized by a ligand-, clathrin- and dynaminindependent mechanism, which does not involve the presence of the AChR-associated protein rapsyn. The small GTPase Rac1 is required: expression of a dominant negative form of Rac1, Rac1N17, abrogates receptor endocytosis. At variance with the endocytic pathway in control CHO cells, the accelerated AChR internalization proceeds even upon disruption of the actin cytoskeleton and does not depend on the cytoskeleton-associated inositol lipid PI(4,5)P2; its sequestration by the PH domain of phospholipase C does not alter endocytosis. AChR internalization under Chol- conditions is furthermore found to require the activity of Arf6 and its effectors Rac1 and phospholipase D. Thus, membrane Chol appears to act as a key homeostatic regulator of cell-surface receptor levels, determining the rate and mechanism of AChR endocytosis
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
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
Eukaryotic Lipids;Ttreasure of regulatory information
Spetses
Grecia
Federation of European Biochemical Societies
International Union for Biochemistry and Molecular Biology
Materia
NICOTINIC RECEPTOR
CHOLESTEROL
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:11336/233376
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cellsBorroni, Maria VirginiaBarrantes, Francisco JoseNICOTINIC RECEPTORCHOLESTEROLLIPID DOMAINShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Stability of the nicotinic acetylcholine receptor (AChR) at the cell surface is critical to the correct functioning of the cholinergic synapse. Cholesterol (Chol) is an essential lipid that modulates AChR levels at the plasmalemma and ion translocation. We have studied the endocytosis of AChR in CHO-K1/A5 cells, a Chinese hamster ovary (CHO) cell line heterologously expressing murine muscle adult-type receptor under different Chol membrane content. Contrary to the norm, endocytosis of cell-surface AChR is accelerated by membrane Chol depletion via a hitherto unknown mechanism. This acceleration is no longer operative when membrane Chol levels are restored. We explored the possible mechanism involved in receptor loss in Choldepleted cells (Chol-). Under such conditions the AChR is internalized by a ligand-, clathrin- and dynaminindependent mechanism, which does not involve the presence of the AChR-associated protein rapsyn. The small GTPase Rac1 is required: expression of a dominant negative form of Rac1, Rac1N17, abrogates receptor endocytosis. At variance with the endocytic pathway in control CHO cells, the accelerated AChR internalization proceeds even upon disruption of the actin cytoskeleton and does not depend on the cytoskeleton-associated inositol lipid PI(4,5)P2; its sequestration by the PH domain of phospholipase C does not alter endocytosis. AChR internalization under Chol- conditions is furthermore found to require the activity of Arf6 and its effectors Rac1 and phospholipase D. Thus, membrane Chol appears to act as a key homeostatic regulator of cell-surface receptor levels, determining the rate and mechanism of AChR endocytosisFil: 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; 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; ArgentinaEukaryotic Lipids;Ttreasure of regulatory informationSpetsesGreciaFederation of European Biochemical SocietiesInternational Union for Biochemistry and Molecular BiologyFederation of European Biochemical Societies2010info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectWorkshopBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/vnd.ms-powerpointapplication/pdfhttp://hdl.handle.net/11336/233376Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells; Eukaryotic Lipids;Ttreasure of regulatory information; Spetses; Grecia; 2010; 13-13CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://web.science.uu.nl/archive/Spetses2010/posters.pdfInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:43:09Zoai:ri.conicet.gov.ar:11336/233376instacron: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 09:43:09.781CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells
title Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells
spellingShingle Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells
Borroni, Maria Virginia
NICOTINIC RECEPTOR
CHOLESTEROL
LIPID DOMAINS
title_short Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells
title_full Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells
title_fullStr Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells
title_full_unstemmed Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells
title_sort Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells
dc.creator.none.fl_str_mv Borroni, Maria Virginia
Barrantes, Francisco Jose
author Borroni, Maria Virginia
author_facet Borroni, Maria Virginia
Barrantes, Francisco Jose
author_role author
author2 Barrantes, Francisco Jose
author2_role author
dc.subject.none.fl_str_mv NICOTINIC RECEPTOR
CHOLESTEROL
LIPID DOMAINS
topic NICOTINIC RECEPTOR
CHOLESTEROL
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 Stability of the nicotinic acetylcholine receptor (AChR) at the cell surface is critical to the correct functioning of the cholinergic synapse. Cholesterol (Chol) is an essential lipid that modulates AChR levels at the plasmalemma and ion translocation. We have studied the endocytosis of AChR in CHO-K1/A5 cells, a Chinese hamster ovary (CHO) cell line heterologously expressing murine muscle adult-type receptor under different Chol membrane content. Contrary to the norm, endocytosis of cell-surface AChR is accelerated by membrane Chol depletion via a hitherto unknown mechanism. This acceleration is no longer operative when membrane Chol levels are restored. We explored the possible mechanism involved in receptor loss in Choldepleted cells (Chol-). Under such conditions the AChR is internalized by a ligand-, clathrin- and dynaminindependent mechanism, which does not involve the presence of the AChR-associated protein rapsyn. The small GTPase Rac1 is required: expression of a dominant negative form of Rac1, Rac1N17, abrogates receptor endocytosis. At variance with the endocytic pathway in control CHO cells, the accelerated AChR internalization proceeds even upon disruption of the actin cytoskeleton and does not depend on the cytoskeleton-associated inositol lipid PI(4,5)P2; its sequestration by the PH domain of phospholipase C does not alter endocytosis. AChR internalization under Chol- conditions is furthermore found to require the activity of Arf6 and its effectors Rac1 and phospholipase D. Thus, membrane Chol appears to act as a key homeostatic regulator of cell-surface receptor levels, determining the rate and mechanism of AChR endocytosis
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
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
Eukaryotic Lipids;Ttreasure of regulatory information
Spetses
Grecia
Federation of European Biochemical Societies
International Union for Biochemistry and Molecular Biology
description Stability of the nicotinic acetylcholine receptor (AChR) at the cell surface is critical to the correct functioning of the cholinergic synapse. Cholesterol (Chol) is an essential lipid that modulates AChR levels at the plasmalemma and ion translocation. We have studied the endocytosis of AChR in CHO-K1/A5 cells, a Chinese hamster ovary (CHO) cell line heterologously expressing murine muscle adult-type receptor under different Chol membrane content. Contrary to the norm, endocytosis of cell-surface AChR is accelerated by membrane Chol depletion via a hitherto unknown mechanism. This acceleration is no longer operative when membrane Chol levels are restored. We explored the possible mechanism involved in receptor loss in Choldepleted cells (Chol-). Under such conditions the AChR is internalized by a ligand-, clathrin- and dynaminindependent mechanism, which does not involve the presence of the AChR-associated protein rapsyn. The small GTPase Rac1 is required: expression of a dominant negative form of Rac1, Rac1N17, abrogates receptor endocytosis. At variance with the endocytic pathway in control CHO cells, the accelerated AChR internalization proceeds even upon disruption of the actin cytoskeleton and does not depend on the cytoskeleton-associated inositol lipid PI(4,5)P2; its sequestration by the PH domain of phospholipase C does not alter endocytosis. AChR internalization under Chol- conditions is furthermore found to require the activity of Arf6 and its effectors Rac1 and phospholipase D. Thus, membrane Chol appears to act as a key homeostatic regulator of cell-surface receptor levels, determining the rate and mechanism of AChR endocytosis
publishDate 2010
dc.date.none.fl_str_mv 2010
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Workshop
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/11336/233376
Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells; Eukaryotic Lipids;Ttreasure of regulatory information; Spetses; Grecia; 2010; 13-13
CONICET Digital
CONICET
url http://hdl.handle.net/11336/233376
identifier_str_mv Cholesterol levels determine AChR endocytic route in CHO-K1/A5 cells; Eukaryotic Lipids;Ttreasure of regulatory information; Spetses; Grecia; 2010; 13-13
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://web.science.uu.nl/archive/Spetses2010/posters.pdf
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/vnd.ms-powerpoint
application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Federation of European Biochemical Societies
publisher.none.fl_str_mv Federation of European Biochemical Societies
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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