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TRANSFERRIN UPTAKE MAY OCCUR THROUGH DETERGENT-RESISTANT MEMBRANE DOMAINS AT THE CYTOPHARYNX OF TRYPANOSOMA CRUZI EPIMASTIGOTE FORMS
Membrana resistente a detergente
Trypanosoma cruzi
Ultraestrutura
Formas de epimastigote
Detergent-resistant membrane
Endocytosis
Trypanosoma cruzi
Ultrastructure
Epimastigote forms
Affilliation
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Ultra-estrutura Celular. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Bioquímica Médica. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Bioquímica Médica. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.
Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Bioquímica Médica. Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Bioquímica Médica. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Instituto de Biologia Molecular do Paraná. Curitiba, PR, Brasil.
Abstract
Uptake of transferrin by epimastigote forms of the protozoan Trypanosoma cruzi occurs mainly through a cytostome/ cytopharynx, via uncoated endocytic vesicles that bud off from the bottom of the cytopharynx. We have here examined whether detergent-resistant membrane (DRM) domains might be involved in this process. Purified whole cell membrane fractions were assayed for cholesterol levels and used in dot blot analyses. Detergent-resistant membrane markers (cholera B toxin and anti-flotillin-1 antibody) presented positive reaction by dot blots in cholesterol-rich/ protein-poor membrane sub-fractions. The positive dot blot fraction was submitted to lipid composition analysis, showing composition similar to that of raft fractions described for other eukaryotic cells. Immunofluorescence assays allowed the localization of punctual positive signal for flotillin-1, matching the precise cytostome/ cytopharynx location. These data were confirmed by immunofluorescence assays with the co-localization of flotillin-1 and the transferrin uptake site. Our data suggest that DRM domains occur and are integrated at the cytostome/ cytopharynx of T. cruzi epimastigotes, being the main route for transferrin uptake.
Keywords in Portuguese
EndocitoseMembrana resistente a detergente
Trypanosoma cruzi
Ultraestrutura
Formas de epimastigote
Keywords
CytopharynxDetergent-resistant membrane
Endocytosis
Trypanosoma cruzi
Ultrastructure
Epimastigote forms
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