Images of mitochondrial UCP 1 in mouse thymocytes using confocal microscopy
Images of mitochondrial UCP 1 in mouse thymocytes using confocal microscopy
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DOI：
10.1016/j.bbabio.2007.10.003
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发表时间：
2008-02-01
影响因子：
4.3
通讯作者：
Porter, Richard K.
中科院分区：
文献类型：
--
作者：
Adams, Alison E.;Hanrahan, Orla;Porter, Richard K.
本研究的目的是证明线粒体解偶联蛋白1（UCP 1）在纯胸腺细胞的组成型表达，用激光扫描共聚焦显微镜图像。为此，我们探测了来自UCP 1敲除小鼠和野生型小鼠的胸腺细胞。用线粒体示踪红测定胸腺细胞中线粒体的位置，用Hoescht染色标记细胞核。我们证明，所有研究的细胞是胸腺细胞，如通过对胸腺细胞表面标志物Thy 1（CD90）具有特异性的单克隆抗体预先偶联到荧光标记（Alexa 448，绿色）所确定的。使用对UCP 1特异的一级肽抗体和二级荧光标记抗体（Alexa 647，洋红色），我们能够证明UCP 1与线粒体相关，在来自UCP 1野生型小鼠的胸腺细胞中，而不是来自UCP 1敲除小鼠的胸腺细胞中。这是第一张证明UCP 1在胸腺细胞线粒体中原位存在的图像，也是第一张清楚地证明UCP 1在棕色脂肪细胞以外的细胞中表达的图像。我们的结论是，小鼠胸腺细胞含有UCP 1在其线粒体。(c)2007 Elsevier B.V.保留所有权利。
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 UCP 1 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. (c) 2007 Elsevier B.V. All rights reserved.