Inositol 1,4,5-trisphosphate receptor determines intracellular Ca²⁺ concentration in Trypanosoma cruzi throughout its life cycle.

ABSTRACT: Regulation of intracellular Ca²⁺ concentration ([Ca²⁺]_i) is vital for eukaryotic organisms. Recently, we identified a Ca²⁺ channel (TcIP ₃R) associated with intracellular Ca²⁺ stores in Trypanosoma cruzi, the parasitic protist that causes Chagas disease. In this study, we measured [Ca²⁺]_i during the parasite life cycle and determined whether TcIP ₃R is involved in the observed variations. Parasites expressing R-GECO1, a red fluorescent, genetically encoded Ca²⁺ indicator for optical imaging that fluoresces when bound to Ca²⁺, were produced. Using these R-GECO1-expressing parasites to measure [Ca²⁺]_i, we found that the [Ca²⁺]_i in epimastigotes was significantly higher than that in trypomastigotes and lower than that in amastigotes, and we observed a positive correlation between TcIP₃R mRNA expression and [Ca²⁺]_i during the parasite life cycle both in vitro and in vivo. We also generated R-GECO1-expressing parasites with TcIP ₃R expression levels that were approximately 65% of wild-type (wt) levels (SKO parasites), and [Ca²⁺]_i in the wt and SKO parasites was compared. The [Ca²⁺]_i in SKO parasites was reduced to approximately 50-65% of that in wt parasites. These results show that TcIP ₃R is the determinant of [Ca²⁺]_i in T. cruzi. Since Ca²⁺ signaling is vital for these parasites, TcIP ₃R is a promising drug target for Chagas disease.

SUBMITTER: Hashimoto M 

PROVIDER: S-EPMC5302059 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature