Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis

Autores
Caruso, Benjamin; Mangiarotti, Agustín; Wilke, Natalia; Perillo, Maria Angelica
Año de publicación
2018
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Lipid Droplets (LD) are intracelular structures consisting on an apolar lipid core - composed mainly of triglycerides (TG) and steryl esters- which is surrounded by a phospholipid and protein monolayer. LDs originate in the ER bilayer, where TG synthesis concludes. The mechanisms underlying TG nucleation, size maduration and budding-off from the ER membrane are a matter of current investigations and the role of dewetting from cytosolic-bilayer interface appears to play a critical role. In order to contrast the nano-sized "blisters" of TG that some authors predict1, here we formed free-standing bilayers by transferring films of a monolayer of mixed phosphatidylcholine(EPC)/TG in coexistence with TG microlenses (i.e. an excluded TG phase floating in the surface). These membranes were characterized by adding them the solvatochromic fluorescent probe Nile Red (NR) and observing them under spectral confocal microscope. Such bilayers exhibit fluorescence emission spectra comparable of bilayers of vesicles with similar composition (POPC and TO). By comparison with literature data and fluorescence spectra of EPC and TG monolayers, the peaks could be assigned to different phases, namely 1) PC membranes (λemmax=630 nm ) bilayer and bilayer) and 2) TG isotropic phase (λemmax=570 nm ). No microscopic structures could be observed at λ emmax=570 nm. Diffusion of NR under this TG phase was characterized using FRAP analysis yielding values (D=2 μm2s ) typical of model bilayer membranes, suggesting that the probe is diffusing in a 2D structure. This system appears appropiate for describing which is the distribution of the TG phase, that is, homogeneously among the intrabilayer space or in nanoscopic "blisters", by evaluating diffusion times obtained by FCS and FRAP.
Fil: Caruso, Benjamin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Mangiarotti, Agustín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Wilke, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Perillo, Maria Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
XLVII Reunión Anual de la Sociedad Argentina de Biofísica
La Plata
Argentina
Sociedad Argentina de Biofísica
Universidad Nacional de La plata. Facultad de Ciencias Médicas
Materia
Biomembranas
Lipid Droplets
Fluorescence Correlation Spectroscopy
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:11336/154225
Acceder
id CONICETDig_a702a07085bca7b0d70d3bbc5fd3dacb
oai_identifier_str oai:ri.conicet.gov.ar:11336/154225
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesisCaruso, BenjaminMangiarotti, AgustínWilke, NataliaPerillo, Maria AngelicaBiomembranasLipid DropletsFluorescence Correlation Spectroscopyhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Lipid Droplets (LD) are intracelular structures consisting on an apolar lipid core - composed mainly of triglycerides (TG) and steryl esters- which is surrounded by a phospholipid and protein monolayer. LDs originate in the ER bilayer, where TG synthesis concludes. The mechanisms underlying TG nucleation, size maduration and budding-off from the ER membrane are a matter of current investigations and the role of dewetting from cytosolic-bilayer interface appears to play a critical role. In order to contrast the nano-sized "blisters" of TG that some authors predict1, here we formed free-standing bilayers by transferring films of a monolayer of mixed phosphatidylcholine(EPC)/TG in coexistence with TG microlenses (i.e. an excluded TG phase floating in the surface). These membranes were characterized by adding them the solvatochromic fluorescent probe Nile Red (NR) and observing them under spectral confocal microscope. Such bilayers exhibit fluorescence emission spectra comparable of bilayers of vesicles with similar composition (POPC and TO). By comparison with literature data and fluorescence spectra of EPC and TG monolayers, the peaks could be assigned to different phases, namely 1) PC membranes (λemmax=630 nm ) bilayer and bilayer) and 2) TG isotropic phase (λemmax=570 nm ). No microscopic structures could be observed at λ emmax=570 nm. Diffusion of NR under this TG phase was characterized using FRAP analysis yielding values (D=2 μm2s ) typical of model bilayer membranes, suggesting that the probe is diffusing in a 2D structure. This system appears appropiate for describing which is the distribution of the TG phase, that is, homogeneously among the intrabilayer space or in nanoscopic "blisters", by evaluating diffusion times obtained by FCS and FRAP.Fil: Caruso, Benjamin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Mangiarotti, Agustín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Wilke, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Perillo, Maria Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaXLVII Reunión Anual de la Sociedad Argentina de BiofísicaLa PlataArgentinaSociedad Argentina de BiofísicaUniversidad Nacional de La plata. Facultad de Ciencias MédicasSociedad Argentina de BiofísicaCelej, Maria Soledad2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/154225Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis; XLVII Reunión Anual de la Sociedad Argentina de Biofísica; La Plata; Argentina; 2018; 75-75978-987-27591-6-2CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/reuniones-cientificas/reunionsab-previas/info:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/xlii-reunion-anual-sab/Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:57:16Zoai:ri.conicet.gov.ar:11336/154225instacron: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:34982025-09-03 09:57:16.275CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis
title Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis
spellingShingle Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis
Caruso, Benjamin
Biomembranas
Lipid Droplets
Fluorescence Correlation Spectroscopy
title_short Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis
title_full Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis
title_fullStr Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis
title_full_unstemmed Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis
title_sort Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis
dc.creator.none.fl_str_mv Caruso, Benjamin
Mangiarotti, Agustín
Wilke, Natalia
Perillo, Maria Angelica
author Caruso, Benjamin
author_facet Caruso, Benjamin
Mangiarotti, Agustín
Wilke, Natalia
Perillo, Maria Angelica
author_role author
author2 Mangiarotti, Agustín
Wilke, Natalia
Perillo, Maria Angelica
author2_role author
author
author
dc.contributor.none.fl_str_mv Celej, Maria Soledad
dc.subject.none.fl_str_mv Biomembranas
Lipid Droplets
Fluorescence Correlation Spectroscopy
topic Biomembranas
Lipid Droplets
Fluorescence Correlation Spectroscopy
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Lipid Droplets (LD) are intracelular structures consisting on an apolar lipid core - composed mainly of triglycerides (TG) and steryl esters- which is surrounded by a phospholipid and protein monolayer. LDs originate in the ER bilayer, where TG synthesis concludes. The mechanisms underlying TG nucleation, size maduration and budding-off from the ER membrane are a matter of current investigations and the role of dewetting from cytosolic-bilayer interface appears to play a critical role. In order to contrast the nano-sized "blisters" of TG that some authors predict1, here we formed free-standing bilayers by transferring films of a monolayer of mixed phosphatidylcholine(EPC)/TG in coexistence with TG microlenses (i.e. an excluded TG phase floating in the surface). These membranes were characterized by adding them the solvatochromic fluorescent probe Nile Red (NR) and observing them under spectral confocal microscope. Such bilayers exhibit fluorescence emission spectra comparable of bilayers of vesicles with similar composition (POPC and TO). By comparison with literature data and fluorescence spectra of EPC and TG monolayers, the peaks could be assigned to different phases, namely 1) PC membranes (λemmax=630 nm ) bilayer and bilayer) and 2) TG isotropic phase (λemmax=570 nm ). No microscopic structures could be observed at λ emmax=570 nm. Diffusion of NR under this TG phase was characterized using FRAP analysis yielding values (D=2 μm2s ) typical of model bilayer membranes, suggesting that the probe is diffusing in a 2D structure. This system appears appropiate for describing which is the distribution of the TG phase, that is, homogeneously among the intrabilayer space or in nanoscopic "blisters", by evaluating diffusion times obtained by FCS and FRAP.
Fil: Caruso, Benjamin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Mangiarotti, Agustín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Wilke, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Perillo, Maria Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
XLVII Reunión Anual de la Sociedad Argentina de Biofísica
La Plata
Argentina
Sociedad Argentina de Biofísica
Universidad Nacional de La plata. Facultad de Ciencias Médicas
description Lipid Droplets (LD) are intracelular structures consisting on an apolar lipid core - composed mainly of triglycerides (TG) and steryl esters- which is surrounded by a phospholipid and protein monolayer. LDs originate in the ER bilayer, where TG synthesis concludes. The mechanisms underlying TG nucleation, size maduration and budding-off from the ER membrane are a matter of current investigations and the role of dewetting from cytosolic-bilayer interface appears to play a critical role. In order to contrast the nano-sized "blisters" of TG that some authors predict1, here we formed free-standing bilayers by transferring films of a monolayer of mixed phosphatidylcholine(EPC)/TG in coexistence with TG microlenses (i.e. an excluded TG phase floating in the surface). These membranes were characterized by adding them the solvatochromic fluorescent probe Nile Red (NR) and observing them under spectral confocal microscope. Such bilayers exhibit fluorescence emission spectra comparable of bilayers of vesicles with similar composition (POPC and TO). By comparison with literature data and fluorescence spectra of EPC and TG monolayers, the peaks could be assigned to different phases, namely 1) PC membranes (λemmax=630 nm ) bilayer and bilayer) and 2) TG isotropic phase (λemmax=570 nm ). No microscopic structures could be observed at λ emmax=570 nm. Diffusion of NR under this TG phase was characterized using FRAP analysis yielding values (D=2 μm2s ) typical of model bilayer membranes, suggesting that the probe is diffusing in a 2D structure. This system appears appropiate for describing which is the distribution of the TG phase, that is, homogeneously among the intrabilayer space or in nanoscopic "blisters", by evaluating diffusion times obtained by FCS and FRAP.
publishDate 2018
dc.date.none.fl_str_mv 2018
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/154225
Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis; XLVII Reunión Anual de la Sociedad Argentina de Biofísica; La Plata; Argentina; 2018; 75-75
978-987-27591-6-2
CONICET Digital
CONICET
url http://hdl.handle.net/11336/154225
identifier_str_mv Evidence of tryglyceride-phase incorporation into artificial bilayers for studying lipid droplets biogenesis; XLVII Reunión Anual de la Sociedad Argentina de Biofísica; La Plata; Argentina; 2018; 75-75
978-987-27591-6-2
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
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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
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Sociedad Argentina de Biofísica
publisher.none.fl_str_mv Sociedad Argentina de Biofísica
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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reponame_str CONICET Digital (CONICET)
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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
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