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dc.contributor.author
Saavedra, Lucas  
dc.contributor.author
Buena Maizón, Héctor  
dc.contributor.author
Barrantes, Francisco Jose  
dc.date.available
2023-10-10T12:49:46Z  
dc.date.issued
2022-09  
dc.identifier.citation
Saavedra, Lucas; Buena Maizón, Héctor; Barrantes, Francisco Jose; Mapping the Nicotinic Acetylcholine Receptor Nanocluster Topography at the Cell Membrane with STED and STORM Nanoscopies; Molecular Diversity Preservation International; International Journal of Molecular Sciences; 23; 18; 9-2022; 1-22  
dc.identifier.issn
1422-0067  
dc.identifier.uri
http://hdl.handle.net/11336/214676  
dc.description.abstract
The cell-surface topography and density of nicotinic acetylcholine receptors (nAChRs) play a key functional role in the synapse. Here we employ in parallel two labeling and two super-resolution microscopy strategies to characterize the distribution of this receptor at the plasma membrane of the mammalian clonal cell line CHO-K1/A5. Cells were interrogated with two targeted techniques (confocal microscopy and stimulated emission depletion (STED) nanoscopy) and single-molecule nanoscopy (stochastic optical reconstruction microscopy, STORM) using the same fluorophore, Alexa Fluor 647, tagged onto either α-bungarotoxin (BTX) or the monoclonal antibody mAb35. Analysis of the topography of nanometer-sized aggregates (“nanoclusters”) was carried out using STORMGraph, a quantitative clustering analysis for single-molecule localization microscopy based on graph theory and community detection, and ASTRICS, an inter-cluster similarity algorithm based on computational geometry. Antibody-induced crosslinking of receptors resulted in nanoclusters with a larger number of receptor molecules and higher densities than those observed in BTX-labeled samples. STORM and STED provided complementary information, STED rendering a direct map of the mesoscale nAChR distribution at distances ~10-times larger than the nanocluster centroid distances measured in STORM samples. By applying photon threshold filtering analysis, we show that it is also possible to detect the mesoscale organization in STORM images.  
dc.format
application/pdf  
dc.language.iso
eng  
dc.publisher
Molecular Diversity Preservation International  
dc.rights
info:eu-repo/semantics/openAccess  
dc.rights.uri
https://creativecommons.org/licenses/by/2.5/ar/  
dc.subject
CHOLESTEROL  
dc.subject
NANOCLUSTERS  
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NANOSCOPY  
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NICOTINIC ACETYLCHOLINE RECEPTOR  
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STED  
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STORM  
dc.subject
SUPER-RESOLUTION MICROSCOPY  
dc.subject.classification
Biofísica  
dc.subject.classification
Ciencias Biológicas  
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CIENCIAS NATURALES Y EXACTAS  
dc.title
Mapping the Nicotinic Acetylcholine Receptor Nanocluster Topography at the Cell Membrane with STED and STORM Nanoscopies  
dc.type
info:eu-repo/semantics/article  
dc.type
info:ar-repo/semantics/artículo  
dc.type
info:eu-repo/semantics/publishedVersion  
dc.date.updated
2023-07-10T11:12:00Z  
dc.journal.volume
23  
dc.journal.number
18  
dc.journal.pagination
1-22  
dc.journal.pais
Suiza  
dc.description.fil
Fil: Saavedra, Lucas. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina  
dc.description.fil
Fil: Buena Maizón, Héctor. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina  
dc.description.fil
Fil: Barrantes, Francisco Jose. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentina  
dc.journal.title
International Journal of Molecular Sciences  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/1422-0067/23/18/10435  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.3390/ijms231810435