Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation

Bibliographic Details
Main Author: Saldanha, C.
Publication Date: 2002
Other Authors: Santos, N. C., Martins-Silva, J.
Format: Article
Language: eng
Source: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Download full: http://hdl.handle.net/10451/20183
Summary: © 2002 Springer-Verlag New York Inc.
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spelling Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculationErythocyte membranevesicleAcetylcholinesteraseMembrane fluidityLipid microdomainsCholesterolDynamic light scattering© 2002 Springer-Verlag New York Inc.An experimental approach has been developed to study human erythrocyte vesiculation, using the fluorescent probes diphenylhexatriene (DPH), trimethylamino-diphenylhexatriene (TMA-DPH) and heptadecyl-hydroxycoumarin (C17-HC). Acetylcholinesterase (AChE) enzyme activity measurements confirmed the presence of exovesicles released from erythrocyte membranes labeled with DPH, TMA-DPH or C17-HC. The fluorescence intensity and anisotropy values obtained showed that the amphiphilic probes TMA-DPH and C17-HC are preferentially incorporated in the exovesicles (when compared with DPH). There is a significant decrease of the cholesterol content of the exovesicle suspensions with time, independently of the fluorescence probe used, reaching undetectable cholesterol levels for the samples incubated for 48 hr. The ratios between the concentration of cholesterol released in the exovesicles after 1 hr incubation with DPH, TMA-DPH or C17-HC and the probe concentration used in the incubation were 84.7, 3.82 and 0.074, respectively. The size of the released vesicles was evaluated by dynamic light scattering spectroscopy. Some hypotheses are proposed that could explain the resemblance and differences between the results obtained for erythrocytes labeled with each probe, considering the present knowledge of membrane vesiculation mechanisms, lipid microdomains (rafts), erythrocyte membrane phospholipid asymmetry and AChE inhibition by TMA-DPH and C17-HC. This work demonstrates that the fluorescent probes DPH, TMA-DPH and C17-HC induce rapid erythrocyte exovesiculation; their use can lead to new methodologies for the study of this still poorly understood mechanism.This work was supported by the Fundação para a Ciência e Tecnologia (FCT) of the Portuguese Ministry of Science, Technology and Higher Education (MCT) and by the Calouste Gulbenkian Foundation (Portugal)SpringerRepositório da Universidade de LisboaSaldanha, C.Santos, N. C.Martins-Silva, J.2015-10-01T11:09:45Z20022002-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10451/20183engJ. Membrane Biol. 190, 75-82 (2002)0022-2631http://dx.doi.org/10.1007/s00232-002-1025-5metadata only accessinfo:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2025-03-17T13:20:53Zoai:repositorio.ulisboa.pt:10451/20183Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-29T02:41:18.644407Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation
title Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation
spellingShingle Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation
Saldanha, C.
Erythocyte membranevesicle
Acetylcholinesterase
Membrane fluidity
Lipid microdomains
Cholesterol
Dynamic light scattering
title_short Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation
title_full Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation
title_fullStr Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation
title_full_unstemmed Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation
title_sort Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation
author Saldanha, C.
author_facet Saldanha, C.
Santos, N. C.
Martins-Silva, J.
author_role author
author2 Santos, N. C.
Martins-Silva, J.
author2_role author
author
dc.contributor.none.fl_str_mv Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Saldanha, C.
Santos, N. C.
Martins-Silva, J.
dc.subject.por.fl_str_mv Erythocyte membranevesicle
Acetylcholinesterase
Membrane fluidity
Lipid microdomains
Cholesterol
Dynamic light scattering
topic Erythocyte membranevesicle
Acetylcholinesterase
Membrane fluidity
Lipid microdomains
Cholesterol
Dynamic light scattering
description © 2002 Springer-Verlag New York Inc.
publishDate 2002
dc.date.none.fl_str_mv 2002
2002-01-01T00:00:00Z
2015-10-01T11:09:45Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
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dc.identifier.uri.fl_str_mv http://hdl.handle.net/10451/20183
url http://hdl.handle.net/10451/20183
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv J. Membrane Biol. 190, 75-82 (2002)
0022-2631
http://dx.doi.org/10.1007/s00232-002-1025-5
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