Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis

Autores
Günther, German; Herlax, Vanesa Silvana; Lillo, M. Pilar; Sandoval Altamirano, Catalina; Belmar, Libnny N.; Sánchez, Susana A.
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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 (20 μM 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.
Fil: Günther, German. Universidad de Chile; Chile
Fil: Herlax, Vanesa Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina
Fil: Lillo, M. Pilar. Consejo Superior de Investigaciones Científicas. Instituto de Química Física; España
Fil: Sandoval Altamirano, Catalina. Universidad de Chile; Chile
Fil: Belmar, Libnny N.. Universidad de Concepción; Chile
Fil: Sánchez, Susana A.. Universidad de Concepción; Chile
Materia
FLIM PHASOR
LAURDAN GP
MEMBRANE FLUIDITY
MEMBRANE HETEROGENEITY
SOLUBILIZATION
SPECTRAL PHASOR
SUCROSE ESTER
SURFACTANTS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/95331
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/95331
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysisGünther, GermanHerlax, Vanesa SilvanaLillo, M. PilarSandoval Altamirano, CatalinaBelmar, Libnny N.Sánchez, Susana A.FLIM PHASORLAURDAN GPMEMBRANE FLUIDITYMEMBRANE HETEROGENEITYSOLUBILIZATIONSPECTRAL PHASORSUCROSE ESTERSURFACTANTShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The 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 (20 μM 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.Fil: Günther, German. Universidad de Chile; ChileFil: Herlax, Vanesa Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Lillo, M. Pilar. Consejo Superior de Investigaciones Científicas. Instituto de Química Física; EspañaFil: Sandoval Altamirano, Catalina. Universidad de Chile; ChileFil: Belmar, Libnny N.. Universidad de Concepción; ChileFil: Sánchez, Susana A.. Universidad de Concepción; ChileElsevier Science2018-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/95331Günther, German; Herlax, Vanesa Silvana; Lillo, M. Pilar; Sandoval Altamirano, Catalina; Belmar, Libnny N.; et al.; Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 161; 1-2018; 375-3850927-7765CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.colsurfb.2017.10.068info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0927776517307348info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:00:11Zoai:ri.conicet.gov.ar:11336/95331instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-10 13:00:11.484CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis
title Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis
spellingShingle Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis
Günther, German
FLIM PHASOR
LAURDAN GP
MEMBRANE FLUIDITY
MEMBRANE HETEROGENEITY
SOLUBILIZATION
SPECTRAL PHASOR
SUCROSE ESTER
SURFACTANTS
title_short Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis
title_full Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis
title_fullStr Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis
title_full_unstemmed Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis
title_sort Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis
dc.creator.none.fl_str_mv Günther, German
Herlax, Vanesa Silvana
Lillo, M. Pilar
Sandoval Altamirano, Catalina
Belmar, Libnny N.
Sánchez, Susana A.
author Günther, German
author_facet Günther, German
Herlax, Vanesa Silvana
Lillo, M. Pilar
Sandoval Altamirano, Catalina
Belmar, Libnny N.
Sánchez, Susana A.
author_role author
author2 Herlax, Vanesa Silvana
Lillo, M. Pilar
Sandoval Altamirano, Catalina
Belmar, Libnny N.
Sánchez, Susana A.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv FLIM PHASOR
LAURDAN GP
MEMBRANE FLUIDITY
MEMBRANE HETEROGENEITY
SOLUBILIZATION
SPECTRAL PHASOR
SUCROSE ESTER
SURFACTANTS
topic FLIM PHASOR
LAURDAN GP
MEMBRANE FLUIDITY
MEMBRANE HETEROGENEITY
SOLUBILIZATION
SPECTRAL PHASOR
SUCROSE ESTER
SURFACTANTS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv 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 (20 μM 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.
Fil: Günther, German. Universidad de Chile; Chile
Fil: Herlax, Vanesa Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina
Fil: Lillo, M. Pilar. Consejo Superior de Investigaciones Científicas. Instituto de Química Física; España
Fil: Sandoval Altamirano, Catalina. Universidad de Chile; Chile
Fil: Belmar, Libnny N.. Universidad de Concepción; Chile
Fil: Sánchez, Susana A.. Universidad de Concepción; Chile
description 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 (20 μM 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.
publishDate 2018
dc.date.none.fl_str_mv 2018-01
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/95331
Günther, German; Herlax, Vanesa Silvana; Lillo, M. Pilar; Sandoval Altamirano, Catalina; Belmar, Libnny N.; et al.; Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 161; 1-2018; 375-385
0927-7765
CONICET Digital
CONICET
url http://hdl.handle.net/11336/95331
identifier_str_mv Günther, German; Herlax, Vanesa Silvana; Lillo, M. Pilar; Sandoval Altamirano, Catalina; Belmar, Libnny N.; et al.; Study of rabbit erythrocytes membrane solubilization by sucrose monomyristate using laurdan and phasor analysis; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 161; 1-2018; 375-385
0927-7765
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.colsurfb.2017.10.068
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0927776517307348
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1842979864617943040
score 12.993085

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