Regulation of ecto‐NTPDase I gene expression in Trypanosoma cruzi (974.7)

The ecto‐NTPDase I is an apyrase located on the outer surface of Trypanosoma cruzi plasma membrane, and previous data suggest that it possess a role to the infectivity and virulence of the parasite. In this study, we evaluate the mRNA levels for the Ecto‐NTPDase I by RT‐qPCR in distinct T. cruzi strains/clones and distinct evolutive forms. The Real Time PCR assays were performed with primers designed to amplify a 111bp sequence in the ecto‐NTPDase I T. cruzi gene and, as endogenous controls, a 268bp sequence in the calmodulin T. cruzi gene and a 100bp sequence in the GAPDH T.cruzi gene were used. We observed that the strains/ clones Cl‐Brener, Dm28c, 4167, LL014, 3663 and Y presented mRNA expression levels, respectively, 8,21±1,62; 7,17±1,51; 1,26±0,18; 1,20±0; 1,06±0,04; 0,94±0,08, times higher than the CL‐14 strain. Surprisingly, we observed that the epimastigote and amastigote presented expression levels, respectively, 14,35±2,04 and 10,23±1,16 times higher than the trypomastigote form. Moreover, following the parasite growth curve, we also observed that the expression of ecto‐NTPDase I raises in a time‐dependent manner, which could be related to a parasite response to a purine starvation in the cultivation medium. So far, our results suggest that the ecto‐NTPDase I expression is regulated at mRNA level, which is distinct between T. cruzi strains and evolutive forms. Grant Funding Source : Supported by CNPq and FAPERJ