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

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, w...

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Detalles Bibliográficos
Autores principales: Kamerbeek, Constanza B., Borroni, María Virginia, Pediconi, María F., Sato, Satoshi B., Kobayashi, Toshihide, Barrantes, Francisco José
Formato: Artículo
Lenguaje:Inglés
Publicado: Elsevier B.V. 2019
Materias:
MEDICINA
RECEPTORES
MEMBRANAS CELULARES
COLESTEROL
ANTICUERPOS
Acceso en línea:https://repositorio.uca.edu.ar/handle/123456789/8752
Aporte de:
Repositorio Institucional de la Universidad Católica Argentina (UCA) de Universidad Católica Argentina
id I33-R139123456789-8752
record_format dspace
institution Universidad Católica Argentina
institution_str I-33
repository_str R-139
collection Repositorio Institucional de la Universidad Católica Argentina (UCA)
language Inglés
topic MEDICINA
RECEPTORES
MEMBRANAS CELULARES
COLESTEROL
ANTICUERPOS
spellingShingle MEDICINA
RECEPTORES
MEMBRANAS CELULARES
COLESTEROL
ANTICUERPOS
Kamerbeek, Constanza B.
Borroni, María Virginia
Pediconi, María F.
Sato, Satoshi B.
Kobayashi, Toshihide
Barrantes, Francisco José
Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
topic_facet MEDICINA
RECEPTORES
MEMBRANAS CELULARES
COLESTEROL
ANTICUERPOS
description 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.
format Artículo
author Kamerbeek, Constanza B.
Borroni, María Virginia
Pediconi, María F.
Sato, Satoshi B.
Kobayashi, Toshihide
Barrantes, Francisco José
author_facet Kamerbeek, Constanza B.
Borroni, María Virginia
Pediconi, María F.
Sato, Satoshi B.
Kobayashi, Toshihide
Barrantes, Francisco José
author_sort Kamerbeek, Constanza B.
title Antibody-induced acetylcholine receptor clusters inhabit liquid-ordered and liquid-disordered domains
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
publisher Elsevier B.V.
publishDate 2019
url https://repositorio.uca.edu.ar/handle/123456789/8752
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bdutipo_str Repositorios
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