Fluorescent probes DPH, TMA-DPH and C17-HC induce erythrocyte exovesiculation
Main Author: | |
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Publication Date: | 2002 |
Other Authors: | , |
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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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 |
status_str |
publishedVersion |
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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metadata only access info:eu-repo/semantics/openAccess |
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Springer |
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Springer |
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