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Images of mitochondrial UCP 1 in mouse thymocytes using confocal microscopy

2007; Elsevier BV; Volume: 1777; Issue: 2 Linguagem: Inglês

10.1016/j.bbabio.2007.10.003

1879-2650

A.E. Adams, Orla Hanrahan, Derek P. Nolan, H. Paul Voorheis, Padraic G. Fallon, Richard K. Porter,

Adipokines, Inflammation, and Metabolic Diseases

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.