Evidence map›Paper›PMID 32566788›Full record

ArticleHeliyon2020

Fluorescent redox-dependent labeling of lipid droplets in cultured cells by reduced phenazine methosulfate.

Juan C Stockert, María C Carou, Adriana G Casas, María C García Vior, Sergio D Ezquerra Riega, María M Blanco, Jesús Espada, Alfonso Blázquez-Castro, Richard W Horobin, Daniel M Lombardo

Open access · goldAbstract read
In one paragraph

Article in Heliyon, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
0.6field-weighted citation impact, top 37% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed, 7 citations in OpenAlex.

  1. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors at 5 institutions in 4 countries.

Juan C StockertUniversidad de Buenos Aires, Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal, Cátedra de Histología y Embriología, Buenos Aires, C1427CWO, Argentina.
María C CarouUniversidad de Buenos Aires, Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal, Cátedra de Histología y Embriología, Buenos Aires, C1427CWO, Argentina.
Adriana G CasasCentro de Investigaciones sobre Porfirinas y Porfirias, Hospital de Clínicas, Universidad de Buenos Aires, CONICET, C1120AAF, Argentina.
María C García ViorUniversidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Departamento de Química Orgánica, C1113AAD, CABA, Argentina.
Sergio D Ezquerra RiegaUniversidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Departamento de Química Orgánica, C1113AAD, CABA, Argentina.
María M BlancoUniversidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Departamento de Química Orgánica, C1113AAD, CABA, Argentina.
Jesús EspadaExperimental Dermatology and Skin Biology Group, Ramón y Cajal Institute for Health Research, Ramón y Cajal University Hospital, 28034, Madrid, Spain.
Alfonso Blázquez-CastroDepartamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Madrid, 28049, Spain.
Richard W HorobinChemical Biology and Precision Synthesis, School of Chemistry, University of Glasgow, Glasgow, G12 8QQ, Scotland, UK.
Daniel M LombardoUniversidad de Buenos Aires, Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal, Cátedra de Histología y Embriología, Buenos Aires, C1427CWO, Argentina.
Universidad de Buenos Aires · ARConsejo Nacional de Investigaciones Científicas y Técnicas · ARInstituto Cajal · ESUniversidad Autónoma de Madrid · ESUniversity of Glasgow · GB

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

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.

Indexed as

BiochemistryBiological sciencesBiomoleculesCell biologyCell cultureFluorescent labelingLipid dropletsPMSQSARRedox reagents

Identifiers

PMID32566788
PMCPMC7298651
OpenAlexW3034330898

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.