Clathrin expression in Trypanosoma cruzi
1 Laboratory of Cell Biology, Carlos Chagas Institute, Fiocruz, Rua Professor Algacyr Munhoz Mader 3775, 81350-010 Curitiba, PR, Brazil
2 Laboratory of Molecular Biology of Trypanosomatids, Carlos Chagas Institute, Fiocruz-PR, 81350-010 Curitiba, PR, Brazil
3 Department of General Biology, State University of Ponta Grossa, 84030-900 Ponta Grossa, PR, Brazil
BMC Cell Biology 2014, 15:23 doi:10.1186/1471-2121-15-23Published: 19 June 2014
Clathrin-mediated vesicular trafficking, the mechanism by which proteins and lipids are transported between membrane-bound organelles, accounts for a large proportion of import from the plasma membrane (endocytosis) and transport from the trans-Golgi network towards the endosomal system. Clathrin-mediated events are still poorly understood in the protozoan Trypanosoma cruzi, the causative agent of Chagas disease in Latin America. In this study, clathrin heavy (TcCHC) and light (TcCLC) chain gene expression and protein localization were investigated in different developmental forms of T. cruzi (epimastigotes, trypomastigotes and amastigotes), using both polyclonal and monoclonal antibodies raised against T. cruzi recombinant proteins.
Analysis by confocal microscopy revealed an accumulation of TcCHC and TcCLC at the cell anterior, where the flagellar pocket and Golgi complex are located. TcCLC partially colocalized with the Golgi marker TcRAB7-GFP and with ingested albumin, but did not colocalize with transferrin, a protein mostly ingested via uncoated vesicles at the cytostome/cytopharynx complex.
Clathrin heavy and light chains are expressed in T. cruzi. Both proteins typically localize anterior to the kinetoplast, at the flagellar pocket and Golgi complex region. Our data also indicate that in T. cruzi epimastigotes clathrin-mediated endocytosis of albumin occurs at the flagellar pocket, while clathrin-independent endocytosis of transferrin occurs at the cytostome/cytopharynx complex.