Abstract
The insect-transmitted protozoan parasite Trypanosoma cruzi experiences changes in nutrient availability and rate of flux through different metabolic pathways across its life cycle. The species encompasses much genetic diversity of both the nuclear and mitochondrial genomes among isolated strains. The genetic or expression variation of both genomes are likely to impact metabolic responses to environmental stimuli, and even steady state metabolic function, among strains. To begin formal characterization these differences, we compared aspects of metabolism between genetically similar strains CL Brener and Tulahuen with less similar Esmeraldo and Sylvio X10 strains in a culture environment. Epimastigotes of all strains took up glucose at similar rates. However, the degree of medium acidification that could be observed when glucose was absent from the medium varied by strain, indicating potential differences in excreted metabolic byproducts. Our main focus was differences related to electron transport chain function. We observed differences in ATP-coupled respiration and maximal respiratory capacity, mitochondrial membrane potential, and mitochondrial morphology between strains, despite the fact that abundances of two nuclear-encoded proteins of the electron transport chain are similar between strains. RNA sequencing reveals strain-specific differences in abundances of mRNAs encoding proteins of the respiratory chain but also other metabolic processes. From these differences in metabolism and mitochondrial phenotypes we have generated tentative models for the differential metabolic fluxes or differences in gene expression that may underlie these results.
Highlights
Fundamental biochemical and gene expression pathways of the disease-causing, insect transmitted trypanosomatids are often deciphered in the model organism Trypanosoma brucei
Replicating insect stage T. cruzi cells from strains CL Brener, Tulahuen cl98 (ATCC1 50829), Sylvio X10 (ATCC1 50823) and Esmeraldo cl3 (ATCC1 50794) were cultured in Liver Infusion Tryptose (LIT) medium [30] supplemented with 10% FBS and 20 μg/ml hemin (Sigma H9039) in 27 ̊C, 5% CO2 incubators
A more challenging restricted medium (RPMI) contains a defined, and narrower range of nutrient sources and other additives and was supplemented with very limited FBS compared to normal medium conditions
Summary
Fundamental biochemical and gene expression pathways of the disease-causing, insect transmitted trypanosomatids are often deciphered in the model organism Trypanosoma brucei. Gene expression and flux through major metabolic pathways are known to dramatically differ between T. brucei proliferative insect and mammalian life stages [1,2,3,4]. Gene expression and mitochondrial metabolism in Trypanosoma cruzi strains study design, data collection and analysis, decision to publish, or preparation of the manuscript
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