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Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis.
- Source :
-
Colloids and surfaces. B, Biointerfaces [Colloids Surf B Biointerfaces] 2018 Jan 01; Vol. 161, pp. 375-385. Date of Electronic Publication: 2017 Nov 01. - Publication Year :
- 2018
-
Abstract
- The study of surfactant and bio membranes interaction is particularly complex due to the diversity in lipid composition and the presence of proteins in natural membranes. Even more difficult is the study of this interaction in vivo since cellular damage may complicate the interpretation of the results, therefore for most of the studies in this field either artificial or model systems are used. One of the model system most used to study biomembranes are erythrocytes due to their relatively simple structure (they lack nuclei and organelles having only the plasma membrane), their convenient experimental manipulation and availability. In this context, we used rabbit erythrocytes as a model membrane and Laurdan (6-lauroyl-2-dimethylaminonaphthalene) as the fluorescent probe to study changes promoted in the membrane by the interaction with the sucrose monoester of myristic acid, β-d-fructofuranosyl-6-O-myristoyl-α-d-glucopyranoside (MMS). Surfactant and erythrocytes interaction was studied by measuring hemoglobin release and the changes in water content in the membrane sensed by Laurdan. Using two-photon excitation, three types of measurements were performed: Generalized Polarization (analyzed as average GP values), Fluorescence Lifetime Imaging, FLIM (analyzed using phasor plots) and Spectral imaging (analyzed using spectral phasor). Our data indicate that at sublytical concentration of surfactant (20μM MMS), there is a decrease of about 35% in erythrocytes size, without changes in Laurdan lifetime or emission spectra. We also demonstrate that as hemolysis progress, Laurdan lifetime increased due to the decrease in hemoglobin (strong quencher of Laurdan emission) content inside the erythrocytes. Under these conditions, Laurdan spectral phasor analyses can extract the information on the water content in the membrane in the presence of hemoglobin. Our results indicate an increase in membrane fluidity in presence of MMS.<br /> (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Subjects :
- 2-Naphthylamine chemistry
2-Naphthylamine metabolism
Animals
Erythrocyte Membrane chemistry
Erythrocyte Membrane drug effects
Erythrocytes drug effects
Fluorescent Dyes chemistry
Fluorescent Dyes metabolism
Hemoglobins metabolism
Hemolysis
Laurates chemistry
Membrane Fluidity drug effects
Pulmonary Surfactants pharmacology
Rabbits
Solubility
Water metabolism
2-Naphthylamine analogs & derivatives
Erythrocyte Membrane metabolism
Erythrocytes metabolism
Laurates metabolism
Myristic Acid metabolism
Sucrose metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1873-4367
- Volume :
- 161
- Database :
- MEDLINE
- Journal :
- Colloids and surfaces. B, Biointerfaces
- Publication Type :
- Academic Journal
- Accession number :
- 29102849
- Full Text :
- https://doi.org/10.1016/j.colsurfb.2017.10.068