Metabolic studies by 1H NMR of different forms of Trypanosoma cruzi as obtained by 'in vitro' culture.
By culturing Trypanosoma cruzi epimastigotes in modified Grace's medium with 10% foetal bovine serum, a significant quantity of metacyclic forms could be obtained. Transformation was observed after 8 days of culture, with metacyclic forms reaching 75%. Cultured Vero cells were infected with metacyclic forms and maintained until free-amastigote forms were obtained. Additionally, amastigote-like forms could be obtained by subjecting metacyclic cultures to heat shock. Parasites were grown with glucose as the major carbon source. The metabolites produced and excreted during culture were identified by difference proton nuclear magnetic resonance spectroscopy and quantified by enzymatic methods. The final products of glucose catabolism differed not only quantitatively but also qualitatively for the three major life-cycle stages of T. cruzi. The end products of metabolism produced by epimastigote forms were mainly acetate and pyruvate and, to a lesser extend, L-alanine and ethanol. Differences between epimastigotes and metacyclic forms were only quantitative. However, free amastigotes as well as amastigote-like forms, excreted acetate, glycerol, and pyruvate and to a lesser extent succinate, but no L-alanine or ethanol.
Sanchez-Moreno
M
M
Institute of Biotechnology, Faculty of Sciences, University of Granada, Spain.
Fernandez-Becerra
M C
MC
Castilla-Calvente
J J
JJ
Osuna
A
A
D000818:Animals; D001798:Blood Proteins; D002417:Cattle; D048429:Cell Size / Q000502:physiology; D002522:Chlorocebus aethiops; D003470:Culture Media; D003903:Deuterium; D006358:Hot Temperature; D009682:Magnetic Resonance Spectroscopy; D014349:Trypanosoma cruzi / Q000254:growth & development / Q000378:metabolism* / Q000469:parasitology; D014709:Vero Cells / Q000469:parasitology
D001798:Blood Proteins; D003470:Culture Media; D003903:Deuterium