Mariotini-Moura C.,Federal University of Viçosa |
Mariotini-Moura C.,Institute Nacional Of Biotecnologia Estrutural E Quimica Medicinal Em Doencas Infecciosas Inbeqmedi |
Bastos M.S.E.,Federal University of Viçosa |
Bastos M.S.E.,Institute Nacional Of Biotecnologia Estrutural E Quimica Medicinal Em Doencas Infecciosas Inbeqmedi |
And 26 more authors.
Acta Tropica | Year: 2014
Previous work has suggested that Trypanosoma cruzi diphosphohydrolase 1 (TcNTPDase-1) may be involved in the infection of mammalian cells and serve as a potential target for rational drug design. In this work, we produced recombinant TcNTPDase-1 and evaluated its nucleotidase activity, cellular localization and role in parasite adhesion to mammalian host cells. TcNTPDase-1 was able to utilize a broad range of triphosphate and diphosphate nucleosides. The enzyme's Km for ATP (0.096mM) suggested a capability to influence the host's ATP-dependent purinergic signaling. The use of specific polyclonal antibodies allowed us to confirm the presence of TcNTPDase-1 at the surface of parasites by confocal and electron microscopy. In addition, electron microscopy revealed that TcNTPDase-1 was also found in the flagellum, flagellum insertion region, kinetoplast, nucleus and intracellular vesicles. The presence of this enzyme in the flagellum insertion region and vesicles suggests that it may have a role in nutrient acquisition, and the widespread distribution of TcNTPDase-1 within the parasite suggests that it may be involved in other biological process. Adhesion assays using anti-TcNTPDase-1 polyclonal antibodies as a blocker or purified recombinant TcNTPDase-1 as a competitor revealed that the enzyme has a role in parasite-host cell adhesion. These data open new frontiers to future studies on this specific parasite-host interaction and other unknown functions of TcNTPDase-1 related to its ubiquitous localization. © 2013 The Authors.
Maia-Brigagao C.,Federal University of Rio de Janeiro |
Maia-Brigagao C.,Instituto Nacional Of Biologia Estrutural E Bioimagem |
Gadelha A.P.R.,Instituto Nacional Of Biologia Estrutural E Bioimagem |
Gadelha A.P.R.,Brazilian National Institute of Technology |
And 3 more authors.
Microscopy and Microanalysis | Year: 2013
Giardia duodenalis is a protozoan parasite that causes intestinal disorders. The trophozoites present four pairs of flagella. Here we further analyze the structural organization of the anterior flagella associated structures of G. duodenalis. High resolution scanning electron microscopy of detergent-extracted trophozoites revealed novel aspects of the interaction of the anterior flagella axonemes with the marginal plates. Images of the marginal plates showed that it was located in the anterior region of the parasite, above the crossing point of the anterior flagella axonemes toward the periphery of the cell. Two well distinguished structures were seen associated with the anterior flagella. The first one corresponds to the dense rods, located just below the axoneme. The second one is a system of filaments located in the upper portion of the flagellum, facing the marginal plates and connecting these two structures. The thickness of the filaments is around 18 nm and they are spaced at intervals of 4-32 nm (average 18 nm). The length of the filaments may vary from 33 to 240 nm. We suggest that this filamentous structure of Giardia may help the dynamics and behavior of the anterior flagella of trophozoites during protozoan motility and adhesion, providing favorable conditions for the establishment of parasitism. © Microscopy Society of America 2013.
Girard-Dias W.,Federal University of Rio de Janeiro |
Girard-Dias W.,Instituto Nacional Of Biologia Estrutural E Bioimagem |
Alcantara C.L.,Federal University of Rio de Janeiro |
Alcantara C.L.,Instituto Nacional Of Biologia Estrutural E Bioimagem |
And 8 more authors.
Histochemistry and Cell Biology | Year: 2012
The structural organization of Trypanosoma cruzi has been intensely investigated by different microscopy techniques. At the electron microscopy level, bi-dimensional analysis of thin sections of chemically fixed cells has been one of the most commonly used techniques, despite the known potential of generating artifacts during chemical fixation and the subsequent steps of sample preparation. In contrast, more sophisticated and elaborate techniques, such as cryofixation followed by freeze substitution that are known to preserve the samples in a more close-to-native state, have not been widely applied to T. cruzi. In addition, the 3D characterization of such cells has been carried out mostly using 3D reconstruction from serial sections, currently considered a low resolution technique when compared to electron tomography (ET). In this work, we re-visited the 3D ultrastructure of T. cruzi using a combination of two approaches: (1) analysis of both conventionally processed and cryofixed and freeze substituted cells and (2) 3D reconstruction of large volumes by serial electron tomography. The analysis of high-pressure frozen and freeze substituted parasites showed novel characteristics in a number of intracellular structures, both in their structure and content. Organelles generally showed a smooth and regular morphology in some cases presenting a characteristic electron dense content. Ribosomes and new microtubule sets showed an unexpected localization in the cell body. The improved preservation and imaging in 3D of T. cruzi cells using cryopreparation techniques has revealed some novel aspects of the ultrastructural organization of this parasite. © Springer-Verlag 2012.