Study ofTrypanosoma cruziepimastigote cell death by NMR-visible mobile lipid analysis

Abstract

SUMMARYCell death mechanisms inTrypanosoma cruzihave 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/CH3integration from the visible mobile lipids (VML). Surprisingly, this versatile non-invasive spectroscopy technique has never been employed with this purpose inT. cruzi. In the present study it is shown that under different parasite death-conditions the ratio CH2/CH3varied drastically. Thus,T. cruziepimastigotes in apoptotic conditions increase significantly this ratio while in necrotic as well as in autophagic situations the parasites maintain the VML, CH2/CH3ratio, 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 the1H NMR-VML technique is an adequate tool to discriminate differentT. cruzideath pathways.