Transient cholesterol effects on nicotinic acetylcholine receptor cell-surface mobility

Abstract: To what extent do cholesterol-rich lipid platforms modulate the supramolecular organization of the nicotinic acetylcholine receptor (AChR)? To address this question, the dynamics of AChR particles at high density and its cholesterol dependence at the surface of mammalian cells were studied...

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Autores principales: Almarza, Gonzalo, Sánchez, Francisco, Barrantes, Francisco José
Formato: Artículo
Lenguaje:Inglés
Publicado: Public Library of Science 2019
Materias:
MEDICINA
COLESTEROL
RECEPTORES
NEUROTRANSMISORES
MEMBRANAS CELULARES
Acceso en línea:https://repositorio.uca.edu.ar/handle/123456789/8751
Aporte de:
Repositorio Institucional de la Universidad Católica Argentina (UCA) de Universidad Católica Argentina
id I33-R139123456789-8751
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
COLESTEROL
RECEPTORES
NEUROTRANSMISORES
MEMBRANAS CELULARES
spellingShingle MEDICINA
COLESTEROL
RECEPTORES
NEUROTRANSMISORES
MEMBRANAS CELULARES
Almarza, Gonzalo
Sánchez, Francisco
Barrantes, Francisco José
Transient cholesterol effects on nicotinic acetylcholine receptor cell-surface mobility
topic_facet MEDICINA
COLESTEROL
RECEPTORES
NEUROTRANSMISORES
MEMBRANAS CELULARES
description Abstract: To what extent do cholesterol-rich lipid platforms modulate the supramolecular organization of the nicotinic acetylcholine receptor (AChR)? To address this question, the dynamics of AChR particles at high density and its cholesterol dependence at the surface of mammalian cells were studied by combining total internal reflection fluorescence microscopy and single-particle tracking. AChR particles tagged with a monovalent ligand, fluorescent α-bungarotoxin (αBTX), exhibited two mobile pools: i) a highly mobile one undergoing simple Brownian motion (16%) and ii) one with restricted motion (∼50%), the rest being relatively immobile (∼44%). Depletion of membrane cholesterol by methyl-α-cyclodextrin increased the fraction of the first pool to 22% and 33% after 15 and 40 min, respectively; the pool undergoing restricted motion diminished from 50% to 44% and 37%, respectively. Monoclonal antibody binding results in AChR crosslinking-internalization after 2 h; here, antibody binding immobilized within minutes ∼20% of the totally mobile AChR. This proportion dramatically increased upon cholesterol depletion, especially during the initial 10 min (83.3%). Thus, antibody crosslinking and cholesterol depletion exhibited a mutually synergistic effect, increasing the average lifetime of cell-surface AChRs∼10 s to ∼20 s. The instantaneous (microscopic) diffusion coefficient D2-4 of the AChR obtained from the MSD analysis diminished from ∼0.001 µm2 s(-1) to ∼0.0001-0.00033 µm2 s(-1) upon cholesterol depletion, ∼30% of all particles falling into the stationary mode. Thus, muscle-type AChR exhibits heterogeneous motional regimes at the cell surface, modulated by the combination of intrinsic (its supramolecular organization) and extrinsic (membrane cholesterol content) factors.
format Artículo
author Almarza, Gonzalo
Sánchez, Francisco
Barrantes, Francisco José
author_facet Almarza, Gonzalo
Sánchez, Francisco
Barrantes, Francisco José
author_sort Almarza, Gonzalo
title Transient cholesterol effects on nicotinic acetylcholine receptor cell-surface mobility
title_short Transient cholesterol effects on nicotinic acetylcholine receptor cell-surface mobility
title_full Transient cholesterol effects on nicotinic acetylcholine receptor cell-surface mobility
title_fullStr Transient cholesterol effects on nicotinic acetylcholine receptor cell-surface mobility
title_full_unstemmed Transient cholesterol effects on nicotinic acetylcholine receptor cell-surface mobility
title_sort transient cholesterol effects on nicotinic acetylcholine receptor cell-surface mobility
publisher Public Library of Science
publishDate 2019
url https://repositorio.uca.edu.ar/handle/123456789/8751
work_keys_str_mv AT almarzagonzalo transientcholesteroleffectsonnicotinicacetylcholinereceptorcellsurfacemobility
AT sanchezfrancisco transientcholesteroleffectsonnicotinicacetylcholinereceptorcellsurfacemobility
AT barrantesfranciscojose transientcholesteroleffectsonnicotinicacetylcholinereceptorcellsurfacemobility
bdutipo_str Repositorios
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