Images of mitochondrial UCP 1 in mouse thymocytes using confocal microscopy |
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Authors: | Alison E. Adams Derek N. Nolan Padraic Fallon |
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Affiliation: | a School of Biochemistry and Immunology, Trinity College Dublin, Dublin 2, Ireland b School of Health Science, Trinity College Dublin, St. James' Hospital, Dublin 8, Ireland |
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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. |
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Keywords: | BAT, brown adipose tissue BSA, bovine serum albumin FBS, foetal bovine serum PBS, phosphate buffered saline UCP, uncoupling protein |
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