title: Mitochondria-derived organelles in the diplomonad fish parasite Spironucleus vortens.
abstract: In some eukaryotes, mitochondria have become modified during evolution to yield derived organelles (MDOs) of a similar size (hydrogenosomes), or extremely reduced to produce tiny cellular vesicles (mitosomes). The current study provides evidence for the presence of MDOs in the highly infectious fish pathogen Spironucleus vortens, an organism that produces H₂ and is shown here to have no detectable cytochromes. Transmission electron microscopy (TEM) reveals that S. vortens trophozoites contain electron-dense, membranous structures sometimes with an electron-dense core (200 nm-1 μm), resembling the hydrogenosomes previously described in other protists from habitats deficient in O₂. Confocal microscopy establishes that these organelles exhibit autofluorescence emission spectra similar to flavoprotein constituents previously described for mitochondria and also present in hydrogenosomes. These organelles possess a membrane potential and are labelled by a fluorescently labeled antibody against Fe-hydrogenase from Blastocystis hominis. Heterologous antibodies raised to mitochondrial proteins frataxin and Isu1, also exhibit a discrete punctate pattern of localization in S. vortens; however these labelled structures are distinctly smaller (90-150 nm) than hydrogenosomes as observed previously in other organisms. TEM confirms the presence of double-membrane bounded organelles of this smaller size. In addition, strong background immunostaining occurs in the cytosol for frataxin and Isu1, and labelling by anti-ferredoxin antibody is generally distributed and not specifically localized except for at the anterior polar region. This suggests that some of the functions traditionally attributed to such MDOs may also occur elsewhere. The specialized parasitic life-style of S. vortens may necessitate more complex intracellular compartmentation of redox reactions than previously recognized. Control of infection requires biochemical characterization of redox-related organelles.
authors.affiliation: School of Biosciences, Cardiff University, Cardiff, Wales CF10 3AT, UK.
authors.forename: Coralie O M
authors.initials: CO
authors.lastname: Millet
authors.forename: Catrin F
authors.initials: CF
authors.lastname: Williams
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Hayes
authors.forename: Anthony C
authors.initials: AC
authors.lastname: Hann
authors.forename: Joanne
authors.initials: J
authors.lastname: Cable
authors.forename: David
authors.initials: D
authors.lastname: Lloyd
mesh_terms: D000818:Animals; D016828:Diplomonadida / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure*; D005393:Fish Diseases / Q000469:parasitology; D005398:Fisheries; D005399:Fishes; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D006859:Hydrogen / Q000378:metabolism; D033862:Iron-Binding Proteins / Q000032:analysis / Q000276:immunology; D008564:Membrane Potentials; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D024101:Mitochondrial Proteins / Q000032:analysis / Q000276:immunology; D061848:Optical Imaging; D015388:Organelles / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure*; D013053:Spectrophotometry; D000096923:Frataxin
chemical_list: D005456:Fluorescent Dyes; D006859:Hydrogen; D033862:Iron-Binding Proteins; D024101:Mitochondrial Proteins; D000096923:Frataxin
keywords: Flavoprotein; Frataxin; Hydrogenase; Hydrogenosomes; Isu1; MDO; Mitosomes; PBS; TEM; TMRE; mitochondria-derived organelles; phosphate-buffered saline at pH 7.2; tetramethyl rhodamine ethyl ester; transmission electron microscopy