Trypanosoma cruzi: Flow Cytometric Analysis. I. Analysis of Total DNA/Organism by Means of Mithramycin‐Induced Fluorescence1,2

Dvorak, James A.; Hall, Thomas; Crane, Mark St. J.; Engel, Juan C.; McDaniel, James P.; Uriegas, Rodolfo

doi:10.1111/j.1550-7408.1982.tb05427.x

Cited by 91 publications

(41 citation statements)

References 27 publications

(7 reference statements)

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“…However, the pattern of lectin-binding sites obtained for T. cruzi Dm28c (isolated from opossum) differed from that obtained for the Y strain (isolated from man) (Pereira et al, 1980). This observation is in agreement with the generally described heterogeneity of the parasite (Morel et al, 1980;Dvorak et al, 1980;Engel et al, 1982;Schottelius & Uhlenbruck, 1983;Tibayrenc et al, 1986;Stevens et al, 1988. Aymerich & Goldenberg, 1989).…”

Section: Discussionsupporting

confidence: 81%

Changes in cell-surface carbohydrates of Trypanosoma cruzi during metacyclogenesis under chemically defined conditions

Andrade¹,

Esteves²,

Angluster³

et al. 1991

Journal of General Microbiology

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Highly purified lectins with specificities for receptor molecules containing sialic acid, N-acetylglucosamine (DGlcNAc), N-acetylgalactosamine (D-GalNAc), galactose (D-Gal), mannose-like residues (D-Man) or L-fucose (LFuc), were used to determine changes in cell-surface carbohydrates of the protozoal parasite Trypanosomu cruzi during metacyclogenesis under chemically defined conditions. Of the D-GalNAc-binding lectins, BS-I selectively agglutinated metacyclic trypomastigotes, MPL was selective for replicating epimastigotes, whereas SBA strongly agglutinated all developmental stages of T. cruzi. WGA (sialic acid and/or D-GIcNAc specific) was also reactive with differentiating epimastigotes and metacyclic trypomastigotes but displayed a higher reactivity with replicating epimastigote forms. A progressive decrease in agglutinating activity was observed for jacaline (specific for D-Gal) during the metacyclogenesis process; conversely, a progressive increase in affinity was observed for RCA-I (D-Gabspecific), although the reactivity of other D-Gal-specific lectins (PNA and AxP) was strong at all developmental stages. All developmental stages of T. cruzi were agglutinated by Con A and Lens cufinaris lectins (specific for D-Man-like residues); however, they were unreactive with the L-fucose-binding lectins from Lotus tetrugonolobos and Ufex europueus. These agglutination assays were further confirmed by binding studies using I z51-labelled lectins. Neuraminidase activity was detected in supernatants of cell-free differentiation medium using the PNA hemagglutination test with human A erythrocytes. The most pronounced differences in lectin agglutination activity were observed between replicating and differentiating epimastigotes, suggesting that changes in the composition of accessible cell-surface carbohydrates precede the morphological transformation of epimastigotes into metacyclic trypomastigotes.

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Section: Discussionsupporting

confidence: 81%

Changes in cell-surface carbohydrates of Trypanosoma cruzi during metacyclogenesis under chemically defined conditions

Andrade¹,

Esteves²,

Angluster³

et al. 1991

Journal of General Microbiology

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“…One approach to understanding the relevance of our experimental intra-lineage TcI hybrids to the natural T. cruzi inter-lineage hybrid populations TcIId and TcIIe, is to compare their DNA contents. This approach was pioneered by James Dvorak who used flow cytometric analysis of T. cruzi to show that DNA content varied dramatically between T. cruzi stocks (Dvorak et al 1982). As a basis of this comparative work we prepared a cohort of approximately 50 biological clones of T. cruzi representing all the known genetic lineages and including well-established reference strains.…”

Section: G E N E T I C E X C H a N G E I N T R Y P A N O S O M A C R mentioning

confidence: 99%

The molecular epidemiology and phylogeography of Trypanosoma cruzi and parallel research on Leishmania: looking back and to the future

et al. 2009

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SUMMARYTrypanosoma cruzi is the protozoan agent of Chagas disease, and the most important parasitic disease in Latin America. Protozoa of the genus Leishmania are global agents of visceral and cutaneous leishmaniasis, fatal and disfiguring diseases. In the 1970s multilocus enzyme electrophoresis demonstrated that T. cruzi is a heterogeneous complex. Six zymodemes were described, corresponding with currently recognized lineages, TcI and TcIIa-e – now defined by multiple genetic markers. Molecular epidemiology has substantially resolved the phylogeography and ecological niches of the T. cruzi lineages. Genetic hybridization has fundamentally influenced T. cruzi evolution and epidemiology of Chagas disease. Genetic exchange of T. cruzi in vitro involves fusion of diploids and genome erosion, producing aneuploid hybrids. Transgenic fluorescent clones are new tools to elucidate molecular genetics and phenotypic variation. We speculate that pericardial sequestration plays a role in pathogenesis. Multilocus sequence typing, microsatellites and, ultimately, comparative genomics are improving understanding of T. cruzi population genetics. Similarly, in Leishmania, genetic groups have been defined, including epidemiologically important hybrids; genetic exchange can occur in the sand fly vector. We describe the profound impact of this parallel research on genetic diversity of T. cruzi and Leishmania, in the context of epidemiology, taxonomy and disease control.

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“…Compared with the staining with DAPI alone, the coloration of cytoplasmic RNA with a subsequent PI-stain made the outline of the parasites clearer and increased the brightn ess of the field of vision. Furthermore , differential staining of nuclei with DAPI from cytoplasm with PI enabled one easier discrimination of the parasites than in singl e staining with any of the known fluorescent dyes such as DAPI (3 -5) , mithramycin (12), acridine orange (13,14), rhodamine (15,16), ethidium bromide (17) and auramine (18,19). It should be noted that double staining with DAPI -PI was inadequate to observe mitochondrial DNA except that in T .…”

Section: Discussionmentioning

confidence: 99%

Simple and Rapid Staining for Detection of Entamoeba Cysts and Other Protozoans With Fluorochromes

Kawamoto

Mizuno

Fujioka

et al. 1987

JJMSB

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Trypanosoma cruzi: Flow Cytometric Analysis. I. Analysis of Total DNA/Organism by Means of Mithramycin‐Induced Fluorescence^1,²

Cited by 91 publications

References 27 publications

Changes in cell-surface carbohydrates of Trypanosoma cruzi during metacyclogenesis under chemically defined conditions

Changes in cell-surface carbohydrates of Trypanosoma cruzi during metacyclogenesis under chemically defined conditions

The molecular epidemiology and phylogeography of Trypanosoma cruzi and parallel research on Leishmania: looking back and to the future

Simple and Rapid Staining for Detection of Entamoeba Cysts and Other Protozoans With Fluorochromes

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