title: Isolation and characterization of the mitochondrion-like organelle from Blastocystis hominis.
abstract: Blastocystis hominis in an unusual protozoan parasite of the human intestinal tract. Previous studies have described the presence of mitochondrial-like structures despite the anaerobic nature of the organism. In this study, we describe a simple and rapid technique to isolate and characterize mitochondrion-like organelles (MLO) from B. hominis. The parasite was disrupted using glass beads and the MLO were collected and purified using a sucrose gradient. Negative staining and transmission electron microscopy of the isolated organelles showed mitochondrial-like structures. B. hominis cells were stained with rhodamine 123 and MitoLight to show the presence of transmembrane potential of the MLO. DAPI staining of the cells suggested the presence of DNA in the MLO. Though brief reports have been made in literature, this study is the first to describe a technique for the isolation of the MLO from this organism. Using this technique of isolation, major metabolic functions of the organelle, their associated macromolecules and intra-mitochondrial location can be extensively studied. The role of MLO in this anaerobic protozoan can be widely investigated using this protocol.
authors.affiliation: Laboratory of Molecular and Cellular Parasitology, Department of Microbiology, Faculty of Medicine, National University of Singapore, 5 Science Drive 2, 117597 Singapore.
authors.forename: A M A
authors.initials: AM
authors.lastname: Nasirudeen
authors.forename: Kevin S W
authors.initials: KS
authors.lastname: Tan
mesh_terms: D000818:Animals; D016845:Blastocystis hominis / Q000502:physiology / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry; D007211:Indoles / Q000737:chemistry; D008564:Membrane Potentials / Q000502:physiology; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000502:physiology* / Q000648:ultrastructure; D020112:Rhodamine 123 / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D007211:Indoles; D020112:Rhodamine 123; C007293:DAPI