The sterol composition of Trypanosoma cruzi changes after growth in different culture media and results in different sensitivity to digitonin-permeabilization

Claudia D Rodrigues, Rosana Catisti, Sergio A. Uyemura, Anibal E. Vercesi, Renee Lira, Carlos Rodriguez, Julio A. Urbina, Roberto Docampo

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Respiration, oxidative phosphorylation, and the corresponding changes in membrane potential (ΔΨ) of Trypanosoma cruzi epimastigotes grown either in liver infusion-tryptose (LIT) or brain heart infusion (BHI) culture medium were assayed in situ using digitonin to render their plasma membrane permeable to succinate, ADP, safranine O, and other small molecules. When the cells were permeabilized with 64 μM digitonin, a concentration previously used with epimastigotes, the ability of the cells grown in LIT medium to sustain oxidative phosphorylation was demonstrated by the detection of an oligomycin-sensitive decrease in mitochondrial membrane potential induced by ADP. In contrast, the cells grown in BHI medium were not able to sustain a stable membrane potential and did not respond to ADP addition. Analyses of oxygen consumption by these permeabilized cells indicated that the rate of basal respiration, which was similar in both cell types, was significantly decreased by 64 μM digitonin. Addition of ADP to the permeabilized cells grown in LIT medium promoted an oligomycin-sensitive transition from resting to phosphorylating respiration in contrast to the cells grown in BHI medium, whose respiration decreased steadily and did not respond either to ADP or CCCP. Titration of the cells grown in BHI medium with different digitonin concentrations indicated that their mitochondria have higher sensitivity to digitonin than those grown in LIT medium. Analysis of the sterol composition of epimastigotes grown in the two different media showed a higher percentage of cholesterol in total and mitochondrial extracts of epimastigotes grown in BHI medium as compared to those grown in LIT medium, suggesting the involvement of this sterol in their increased sensitivity to digitonin-permeabilization.

Original languageEnglish
Pages (from-to)588-594
Number of pages7
JournalJournal of Eukaryotic Microbiology
Volume48
Issue number5
StatePublished - Oct 17 2001
Externally publishedYes

Fingerprint

digitonin
Digitonin
Trypanosoma cruzi
Sterols
sterols
Culture Media
culture media
epimastigotes
Adenosine Diphosphate
Growth
heart
breathing
brain
liver
membrane potential
Liver
cells
oligomycin
Brain
Oligomycins

Keywords

  • Cholesterol
  • Membrane potential
  • Mitochondria
  • Oxidative phosphorylation
  • Respiration

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Applied Microbiology and Biotechnology
  • Agricultural and Biological Sciences (miscellaneous)
  • Microbiology

Cite this

The sterol composition of Trypanosoma cruzi changes after growth in different culture media and results in different sensitivity to digitonin-permeabilization. / Rodrigues, Claudia D; Catisti, Rosana; Uyemura, Sergio A.; Vercesi, Anibal E.; Lira, Renee; Rodriguez, Carlos; Urbina, Julio A.; Docampo, Roberto.

In: Journal of Eukaryotic Microbiology, Vol. 48, No. 5, 17.10.2001, p. 588-594.

Research output: Contribution to journalArticle

Rodrigues, Claudia D ; Catisti, Rosana ; Uyemura, Sergio A. ; Vercesi, Anibal E. ; Lira, Renee ; Rodriguez, Carlos ; Urbina, Julio A. ; Docampo, Roberto. / The sterol composition of Trypanosoma cruzi changes after growth in different culture media and results in different sensitivity to digitonin-permeabilization. In: Journal of Eukaryotic Microbiology. 2001 ; Vol. 48, No. 5. pp. 588-594.
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