Images of mitochondrial UCP 1 in mouse thymocytes using confocal microscopy.
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.
Adams
Alison E
AE
School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Ireland.
Hanrahan
Orla
O
Nolan
Derek N
DN
Voorheis
H Paul
HP
Fallon
Padraic
P
Porter
Richard K
RK
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
D007473:Ion Channels; D024101:Mitochondrial Proteins; C000604372:Ucp1 protein, mouse; D000071256:Uncoupling Protein 1