Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes

Detalhes bibliográficos
Autor(a) principal: Batista, Cassiano Martin
Data de Publicação: 2015
Outros Autores: Kessler, Rafael Luis, Eger, Iriane, Soares, Maurilio José
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
DOI: 10.1371/journal.pone.0130165
Texto Completo: https://www.arca.fiocruz.br/handle/icict/22767
Resumo: Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Celular. Curitiba, PR, Brasil.
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spelling Batista, Cassiano MartinKessler, Rafael LuisEger, IrianeSoares, Maurilio José2017-10-04T15:38:18Z2017-10-04T15:38:18Z2015BATISTA, Cassiano Martin et al. Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes. PLoS ONE, v. 10, n. 6, p. 1-18, 2015.1932-6203https://www.arca.fiocruz.br/handle/icict/2276710.1371/journal.pone.0130165porPublic Library of ScienceTrypanosoma cruziChagas DiseaseIntracellular SpaceEndocytosisMammalsEnfermedad de ChagasEspacio IntracelularEndocitosisDoença de ChagasEspaço IntracelularEndocitoseMamíferosTrypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Celular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Genômica Funcional. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Celular. Curitiba, PR, Brasil. / Universidade Estadual de Ponta Grossa. Departamento de Biologia Geral. Ponta Grossa, Paraná, Brazil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Celular. Curitiba, PR, Brasil.Epimastigote forms of Trypanosoma cruzi (the etiologic agent of Chagas disease) internalize and store extracellular macromolecules in lysosome-related organelles (LROs) called reservosomes, which are positive for the cysteine protease cruzipain. Despite the importance of endocytosis for cell proliferation, macromolecule internalization remains poorly understood in the most clinically relevant proliferative form, the intracellular amastigotes found in mammalian hosts. The main obstacle was the lack of a simple method to isolate viable intracellular amastigotes from host cells. In this work we describe the fast and efficient isolation of viable intracellular amastigotes by nitrogen decompression (cavitation), which allowed the analysis of amastigote endocytosis, with direct visualization of internalized cargo inside the cells. The method routinely yielded 5x10(7) amastigotes--with typical shape and positive for the amastigote marker Ssp4--from 5x10(6) infected Vero cells (48 h post-infection). We could visualize the endocytosis of fluorescently-labeled transferrin and albumin by isolated intracellular amastigotes using immunofluorescence microscopy; however, only transferrin endocytosis was detected by flow cytometry (and was also analyzed by western blotting), suggesting that amastigotes internalized relatively low levels of albumin. Transferrin binding to the surface of amastigotes (at 4°C) and its uptake (at 37°C) were confirmed by binding dissociation assays using acetic acid. Importantly, both transferrin and albumin co-localized with cruzipain in amastigote LROs. Our data show that isolated T. cruzi intracellular amastigotes actively ingest macromolecules from the environment and store them in cruzipain-positive LROs functionally related to epimastigote reservosomes.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83084https://www.arca.fiocruz.br/bitstream/icict/22767/1/license.txt783568c2893d2e25a99990b126be1772MD51ORIGINALBatistaTrypanosom.PDFBatistaTrypanosom.PDFapplication/pdf6699272https://www.arca.fiocruz.br/bitstream/icict/22767/2/BatistaTrypanosom.PDF85e4b7086e47ed87e25212c726b28040MD52TEXTBatistaTrypanosom.PDF.txtBatistaTrypanosom.PDF.txtExtracted texttext/plain55588https://www.arca.fiocruz.br/bitstream/icict/22767/3/BatistaTrypanosom.PDF.txtec21c6b3cf6cfdf22b2459e5cb4b4dc7MD53icict/227672018-08-15 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dc.title.pt_BR.fl_str_mv Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes
title Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes
spellingShingle Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes
Batista, Cassiano Martin
Trypanosoma cruzi
Chagas Disease
Intracellular Space
Endocytosis
Mammals
Enfermedad de Chagas
Espacio Intracelular
Endocitosis
Doença de Chagas
Espaço Intracelular
Endocitose
Mamíferos
title_short Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes
title_full Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes
title_fullStr Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes
title_full_unstemmed Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes
title_sort Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes
author Batista, Cassiano Martin
author_facet Batista, Cassiano Martin
Kessler, Rafael Luis
Eger, Iriane
Soares, Maurilio José
author_role author
author2 Kessler, Rafael Luis
Eger, Iriane
Soares, Maurilio José
author2_role author
author
author
dc.contributor.author.fl_str_mv Batista, Cassiano Martin
Kessler, Rafael Luis
Eger, Iriane
Soares, Maurilio José
dc.subject.other.pt_BR.fl_str_mv Trypanosoma cruzi
topic Trypanosoma cruzi
Chagas Disease
Intracellular Space
Endocytosis
Mammals
Enfermedad de Chagas
Espacio Intracelular
Endocitosis
Doença de Chagas
Espaço Intracelular
Endocitose
Mamíferos
dc.subject.en.pt_BR.fl_str_mv Chagas Disease
Intracellular Space
Endocytosis
Mammals
dc.subject.es.pt_BR.fl_str_mv Enfermedad de Chagas
Espacio Intracelular
Endocitosis
dc.subject.decs.pt_BR.fl_str_mv Doença de Chagas
Espaço Intracelular
Endocitose
Mamíferos
description Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Celular. Curitiba, PR, Brasil.
publishDate 2015
dc.date.issued.fl_str_mv 2015
dc.date.accessioned.fl_str_mv 2017-10-04T15:38:18Z
dc.date.available.fl_str_mv 2017-10-04T15:38:18Z
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.citation.fl_str_mv BATISTA, Cassiano Martin et al. Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes. PLoS ONE, v. 10, n. 6, p. 1-18, 2015.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/22767
dc.identifier.issn.pt_BR.fl_str_mv 1932-6203
dc.identifier.doi.none.fl_str_mv 10.1371/journal.pone.0130165
identifier_str_mv BATISTA, Cassiano Martin et al. Trypanosoma cruzi intracellular amastigotes isolated by nitrogen decompression are capable of endocytosis and cargo storage in reservosomes. PLoS ONE, v. 10, n. 6, p. 1-18, 2015.
1932-6203
10.1371/journal.pone.0130165
url https://www.arca.fiocruz.br/handle/icict/22767
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
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