Regulation Of Intracellular Calcium Homeostasis In Trypanosoma Cruzi. Effects Of Calmidazolium And Trifluoperazine

Trypanosoma cruzi epimastigotes maintained an intracellular free calcium concentration of about 0.15 μM, as measured with the fluorescent indicator Fura-2. The maintenance of low [Ca2+]i is energy-dependent since it is disrupted by KCN and FCCP. When the cells were permeabilized with digitonin, the steady-state free Ca2+ concentration in the absence of ATP was about 0.7 μM. The additional presence of ATP resulted in a steady-state level close to 0.1-0.2 μM which compares favorably with the concentration detected in intact cells. Intracellular Ca2+ uptake at high levels of free Ca2+ (1 μM) was due to energy-dependent mitochondrial uptake as indicated by its FCCP-sensitivity. However, as the free Ca2+ concentration was lowered from 1 μM, essentially all uptake was due to the ATP-dependent Ca2+ sequestration by the endoplasmic reticulum as indicated by its stimulation by ATP, and its inhibition by sodium vanadate. 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