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Study of Trypanosoma cruzi epimastigote cell death by NMR-visible mobile lipid analysis

Published online by Cambridge University Press:  05 January 2012

DIEGO BENITEZ
Affiliation:
Grupo de Química Medicinal, Laboratorio de Química Orgánica, Facultad de Ciencias-Facultad de Química, Universidad de la República, Montevideo, Uruguay
HORACIO PEZAROGLO
Affiliation:
Laboratorio de Resonancia Magnética Nuclear, Facultad de Ciencias-Facultad de Química, Universidad de la República, Montevideo, Uruguay
VERÓNICA MARTÍNEZ
Affiliation:
Laboratorio de Resonancia Magnética Nuclear, Facultad de Ciencias-Facultad de Química, Universidad de la República, Montevideo, Uruguay
GABRIELA CASANOVA
Affiliation:
Unidad de Microscopía Electrónica de Transmisión, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay
GONZALO CABRERA
Affiliation:
Programa de Biología Celular y Molecular, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile
NORBEL GALANTI
Affiliation:
Programa de Biología Celular y Molecular, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile
MERCEDES GONZÁLEZ
Affiliation:
Grupo de Química Medicinal, Laboratorio de Química Orgánica, Facultad de Ciencias-Facultad de Química, Universidad de la República, Montevideo, Uruguay
HUGO CERECETTO*
Affiliation:
Grupo de Química Medicinal, Laboratorio de Química Orgánica, Facultad de Ciencias-Facultad de Química, Universidad de la República, Montevideo, Uruguay
*
*Corresponding author: Grupo de Química Medicinal, Laboratorio de Química Orgánica, Facultad de Ciencias-Facultad de Química, Universidad de la República, Iguá 4225, 11400 Montevideo, Uruguay. Tel: +598 2 525 86 18 (7-216). Fax: +598 2 525 0749. E-mail: hcerecet@fq.edu.uy

Summary

Cell death mechanisms in Trypanosoma cruzi have not been disclosed in detail though different conventional techniques have been used in the classification of parasite-cell death type. Nuclear magnetic resonance (NMR) has successfully been used as a tool to evaluate the onset of apoptosis in a number of higher eukaryote-cell models analysing the ratio of CH2/CH3 integration from the visible mobile lipids (VML). Surprisingly, this versatile non-invasive spectroscopy technique has never been employed with this purpose in T. cruzi. In the present study it is shown that under different parasite death-conditions the ratio CH2/CH3 varied drastically. Thus, T. cruzi epimastigotes in apoptotic conditions increase significantly this ratio while in necrotic as well as in autophagic situations the parasites maintain the VML, CH2/CH3 ratio, in normal values. Additionally, other VML markers commonly used in these studies, such as the change in the region of methyl-choline moiety, -N+(CH3)3, exhibited different particular patterns according to the type of cell death. Our results suggest that the 1H NMR-VML technique is an adequate tool to discriminate different T. cruzi death pathways.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2012

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