Heme requirement and intracellular trafficking in Trypanosoma cruzi epimastigotes☆
Section snippets
Materials and methods
Parasites.Trypanosoma cruzi strains Y (CT-IOC-106) and Dm28c (CT-IOC-010) strains were provided by the Trypanosomatid Collection of the Oswaldo Cruz Institute, Fiocruz, Brazil. The protozoa were grown at 28 °C for 7 days in brain–heart infusion medium (BHI), and supplemented with 30 μM hemin (heme-Cl) and 10% fetal calf serum (FCS). Parasite growth was monitored by cell counting in a Neubauer chamber.
Heme, hemoglobin, and globin-derived peptides effects on T. cruzi epimastigotes growth. To
Effects of supplementary hemin on different strains of T. cruzi epimastigotes
We investigated the effects of hemin on the in vitro growth of two strains (Y, Dm28c) of T. cruzi. By varying the hemin concentration from 0 to 1 mM, we observed an increased parasite proliferation in a dose-dependent manner (Fig. 1A and B). Strain Y and clone Dm28c cultivated in BHI medium or BHI medium supplemented with 3, 30, 100, and 1000 μM of heme exhibited respective median generation times of 5.33, 4.07, 3.38, 3.06, 2.66 days (n = 4) and 5.40, 4.61, 4.40, 3.59, 3.30 days (n = 5),
Acknowledgments
We express our gratitude to Dr. Ulysses Lins and Dr. Thais Souto Padron, from Instituto de Microbiologia Prof Paulo de Góes, UFRJ, to allow the mounting of Pd-mP filter on their epifluorescence microscope Zeiss Axioplan 2. We thank Dr. E. Pays and Dr. C. Felu (Brussels) for critical reading of the manuscript.
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This work was supported by FAPERJ, PADCT, CNPq, Howard Hughes Medical Institute (to P.L. Oliveira) and PRONEX (to P.L. Oliveira).
- 1
Present address: Lab. de Microbiologia Celular, Departamento de Micobacterioses, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Brazil.
- 2
These authors contributed equally to this work.