Cellular analysis of host cell infection by different developmental stages of Trypanosoma cruzi.
Trypanosoma cruzi is an obligate intracellular parasite that infects phagocytic and non-phagocytic mammalian cells by a complex process that appears to involve several discrete steps. Even though the infection process was described many years ago, the molecular mechanisms involved remain poorly understood. As fluorescent proteins have proven to be excellent tools for live-cell imaging, we used EGFP- and DsRed1-1-transfected trypomastigotes, amastigotes and epimastigotes to study the infection process in living cells. Contrary to what has been reported, our results showed that epimastigotes are as infective as trypomastigotes and amastigotes. Besides, differences in replication, differentiation and parasite release times were observed among the stages. Our results suggest that the different developmental stages use distinct attachment and invasion mechanisms. We propose that fluorescent-based plasmid expression systems are good models for studying the infection process of intracellular microorganisms and could offers insights about the molecular mechanisms involved.
Florencio-Martínez
Luis
L
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México D.F., Mexico.
Márquez-Dueñas
Claudia
C
Ballesteros-Rodea
Gilberto
G
Martínez-Calvillo
Santiago
S
Manning-Cela
Rebeca
R
D000818:Animals; D003001:Cloning, Molecular; D005347:Fibroblasts / Q000469:parasitology*; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D008018:Life Cycle Stages / Q000502:physiology*; D008164:Luminescent Proteins; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D014162:Transfection; D014349:Trypanosoma cruzi / Q000254:growth & development* / Q000502:physiology
D005456:Fluorescent Dyes; D008164:Luminescent Proteins; C485184:enhanced green fluorescent protein; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins