Study of Trypanosoma cruzi epimastigote cell death by NMR-visible mobile lipid analysis.
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 CH(2)/CH(3) 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 CH(2)/CH(3) 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, CH(2)/CH(3) 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(+)(CH(3))(3), exhibited different particular patterns according to the type of cell death. Our results suggest that the (1)H NMR-VML technique is an adequate tool to discriminate different T. cruzi death pathways.
Grupo de Química Medicinal, Laboratorio de Química Orgánica, Universidad de la República, Montevideo, Uruguay.
Diego
D
Benitez
Horacio
H
Pezaroglo
Verónica
V
Martínez
Gabriela
G
Casanova
Gonzalo
G
Cabrera
Norbel
N
Galanti
Mercedes
M
González
Hugo
H
Cerecetto
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D001343:Autophagy; D016923:Cell Death; D002794:Choline / Q000032:analysis / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008055:Lipids / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009336:Necrosis; D009547:Nifurtimox / Q000494:pharmacology; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000254:growth & development*
D008055:Lipids; D006861:Hydrogen Peroxide; D009547:Nifurtimox; D002794:Choline