Further studies on the organization of the hydrogenosome in Tritrichomonas foetus

The fine structure of the hydrogenosome of Tritrichomonas foetus was analysed using different approaches: routine transmission electron microscopy, quick-freezing techniques followed by freeze-fracture, deep-etching and freeze-substitution, cryo-ultramicrotomy, serial sectioning followed by three-dimensional (3D) reconstruction and cytochemical detection of carbohydrates, Ca++ and phosphates. The presence of two closely apposed unit membranes surrounding the hydrogenosome, as well as its internal vesicle, was shown both in thin sections of well-preserved cells and in freeze-fracture replicas. Analysis of light micrographs, thin serial sections used to 3-D reconstruction and freeze-fracture replicas, show that the hydrogenosome of T. foetus resembles a sphere, but presents a protusion towards the cytoplasm. The vesicle varies in size from organelle to organelle and represent about 8.5% of the volume of the organelle. Based on the fact that the vesicle (a) presents a distinct morphological appearance from the hydrogenosome matrix, (b) was the main site of Ca++-accumulation, (c) presents phosphatase activity and (d) its membrane presents N-acetyl-glucosamine-containing glycoconjugates, as revealed by incubation of cryosections in the presence of gold-labeled WGA, we conclude that it represents a specialized sub-compartment of the hydrogenosome. Freeze-fracture followed by deep etching showed the presence of large number of particles, probably correspondent to macromolecules, within the hydrogenosomal matrix. These structures were not randomly distributed.