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dc.contributor.author
Stockert, Juan C.
dc.contributor.author
Carou, María Clara
dc.contributor.author
Casas, Adriana Gabriela
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Garcia Vior, María Cecilia
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Ezquerra Riega, Sergio Dario
dc.contributor.author
Blanco, María M.
dc.contributor.author
Espada, Jesús
dc.contributor.author
Blázquez Castro, Alfonso
dc.contributor.author
Horobin, Richard W.
dc.contributor.author
Lombardo, Daniel Marcelo
dc.date.available
2021-10-01T13:29:45Z
dc.date.issued
2020-06
dc.identifier.citation
Stockert, Juan C.; Carou, María Clara; Casas, Adriana Gabriela; Garcia Vior, María Cecilia; Ezquerra Riega, Sergio Dario; et al.; Fluorescent redox-dependent labeling of lipid droplets in cultured cells by reduced phenazine methosulfate; Elsevier; Heliyon; 6; 6; 6-2020; 1-6
dc.identifier.issn
2405-8440
dc.identifier.uri
http://hdl.handle.net/11336/142223
dc.description.abstract
Natural and synthetic phenazines are widely used in biomedical sciences. In dehydrogenase histochemistry, phenazine methosulfate (PMS) is applied as a redox reagent for coupling reduced coenzymes to the reduction of tetrazolium salts into colored formazans. PMS is also currently used for cytotoxicity and viability assays of cell cultures using sulfonated tetrazoliums. Under UV (340 nm) excitation, aqueous solutions of the cationic PMS show green fluorescence (λem: 526 nm), whereas the reduced hydrophobic derivative (methyl-phenazine, MPH) shows blue fluorescence (λem: 465 nm). Under UV (365 nm) excitation, cultured cells (LM2, IGROV-1, BGC-1, and 3T3-L1 adipocytes) treated with PMS (5 μg/mL, 30 min) showed cytoplasmic granules with bright blue fluorescence, which correspond to lipid droplets labeled by the lipophilic methyl-phenazine. After formaldehyde fixation blue-fluorescing droplets could be stained with oil red O. Interestingly, PMS-treated 3T3-L1 adipocytes observed under UV excitation 24 h after labeling showed large lipid droplets with a weak green emission within a diffuse pale blue-fluorescing cytoplasm, whereas a strong green emission was observed in small lipid droplets. This fluorescence change from blue to green indicates that reoxidation of methyl-phenazine to PMS can occur. Regarding cell uptake and labeling mechanisms, QSAR models predict that the hydrophilic PMS is not significantly membrane-permeant, so most PMS reduction is expected to be extracellular and associated with a plasma membrane NAD(P)H reductase. Once formed, the lipophilic and blue-fluorescing methyl-phenazine enters live cells and mainly accumulates in lipid droplets. Overall, the results reported here indicate that PMS is an excellent fluorescent probe to investigate labeling and redox dynamics of lipid droplets in cultured cells.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
BIOCHEMISTRY
dc.subject
BIOLOGICAL SCIENCES
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BIOMOLECULES
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CELL BIOLOGY
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CELL CULTURE
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FLUORESCENT LABELING
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LIPID DROPLETS
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PMS
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QSAR
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REDOX REAGENTS
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Biología Celular, Microbiología
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Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
Fluorescent redox-dependent labeling of lipid droplets in cultured cells by reduced phenazine methosulfate
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2021-09-27T15:20:39Z
dc.journal.volume
6
dc.journal.number
6
dc.journal.pagination
1-6
dc.journal.pais
Países Bajos
dc.description.fil
Fil: Stockert, Juan C.. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigación y Tecnología en Reproducción Animal; Argentina
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Fil: Carou, María Clara. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigación y Tecnología en Reproducción Animal; Argentina
dc.description.fil
Fil: Casas, Adriana Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Investigaciones sobre Porfirinas y Porfirias. Universidad de Buenos Aires. Centro de Investigaciones sobre Porfirinas y Porfirias; Argentina
dc.description.fil
Fil: Garcia Vior, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Orgánica; Argentina
dc.description.fil
Fil: Ezquerra Riega, Sergio Dario. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Orgánica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
dc.description.fil
Fil: Blanco, María M.. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Orgánica; Argentina
dc.description.fil
Fil: Espada, Jesús. Universidad Bernardo O'Higgins; Chile
dc.description.fil
Fil: Blázquez Castro, Alfonso. Universidad Autónoma de Madrid. Facultad de Ciencias. Departamento de Biología; España
dc.description.fil
Fil: Horobin, Richard W.. University of Glasgow; Reino Unido
dc.description.fil
Fil: Lombardo, Daniel Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigación y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
dc.journal.title
Heliyon
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S2405844020310264
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.heliyon.2020.e04182
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