title: Images of mitochondrial UCP 1 in mouse thymocytes using confocal microscopy.
abstract: The aim of this study was to demonstrate the constitutive expression of mitochondrial uncoupling protein 1 (UCP 1) in pure thymocytes using laser scanning confocal microscopic imagery. To that end we probed thymocytes from UCP 1 knock-out and wild-type mice. Mitochondrial location in thymocytes was determined using Mitotracker Red and the nucleus was labelled using Hoescht stain. We demonstrate that all cells investigated were thymocytes as determined by a monoclonal antibody specific for the thymocyte surface marker Thy 1 (CD90) pre-coupled to a fluorescent labelled (Alexa 448, green). Using a primary peptide antibody specific to UCP 1, and secondary fluorescently labelled (Alexa 647, magenta) antibody, we were able to demonstrate that UCP 1 is associated with mitochondria in thymocytes from UCP 1 wild-type mice but not thymocytes from UCP1-knock-out mice. These are the first images demonstrating the presence of UCP 1 in thymocyte mitochondria, in situ, and the first to clearly demonstrate UCP 1 expression in cells other than brown adipocytes. We conclude that mouse thymocytes contain UCP 1 in their mitochondria.
authors.affiliation: School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Ireland.
authors.forename: Alison E
authors.initials: AE
authors.lastname: Adams
authors.forename: Orla
authors.initials: O
authors.lastname: Hanrahan
authors.forename: Derek N
authors.initials: DN
authors.lastname: Nolan
authors.forename: H Paul
authors.initials: HP
authors.lastname: Voorheis
authors.forename: Padraic
authors.initials: P
authors.lastname: Fallon
authors.forename: Richard K
authors.initials: RK
authors.lastname: Porter
mesh_terms: D000818:Animals; D005260:Female; D005455:Fluorescent Antibody Technique; D007473:Ion Channels / Q000096:biosynthesis*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D024101:Mitochondrial Proteins / Q000096:biosynthesis*; D013950:Thymus Gland / Q000166:cytology* / Q000378:metabolism; D000071256:Uncoupling Protein 1
chemical_list: D007473:Ion Channels; D024101:Mitochondrial Proteins; C000604372:Ucp1 protein, mouse; D000071256:Uncoupling Protein 1