Trichodinids are ciliated protozoans that reversibly attach to the tegument of marine and freshwater host-organisms via an adhesive disc. In this study, we have used permeabilized cell models of Trichodina pediculus to examine the distribution of centrins, a Ca(2+)-binding protein associated with centrioles and/or contractile filamentous structures in a large number of protists. The previous finding that filamentous material of the adhesive disc comprised a 23-kDa centrin analog suggested that this protein might be a disc-specific isoform. This possibility was explored through immunolabeling methods using two distinct antibodies, anti-ecto-endoplasmic boundary (EEB) and anti-Hscen2 previously shown to react respectively with centrin-based filament networks and with centrioles. Immunofluorescence microscopy showed that anti-EEB reacts with filamentous material of the disc but not with basal bodies. Conversely, anti-Hscen2 cross-reacted with basal bodies but failed to label any type of structure occurring in the disc area. More detailed data on localization of this protein was obtained by immunoelectron microscopy showing gold particles deposits in the lumen of basal bodies. The different patterns revealed by this immunochemical approach suggest that the two protein antigens concerned by this study are distinct centrin isoforms that presumably perform organelle-specific function in the ciliate T. pediculus.
Keywords: Adhesive disc; Centrin; Centrioles; Ciliates; Functional diversification; Trichodina.