In聽vivo labeling of peroxisomes by photoconvertible mEos2 in myelinating glia of mice

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• 作者：Sarah Richert ; S ; ra Kleinecke ; Jenniffer G眉nther ; Florian Schaumburg ; Julia Edgar ; Gerd Ulrich Nienhaus ; Klaus-Armin Nave ; Celia M. Kassmann
• 关键词：Transgenic mouse ; Photoconversion ; Fluorescent peroxisomes ; Oligodendrocytes ; mEos2
• 刊名：Biochimie
• 出版年：March, 2014
• 年：2014
• 卷：98
• 期：Complete
• 页码：127-134
• 全文大小：3385 K

文摘

Mutations of several genes encoding peroxisomal proteins have been associated with human diseases. Some of these display specific white matter abnormalities in the brain, although the affected proteins are ubiquitously expressed. To better understand the etiology of peroxisomal myelin diseases, we aimed to label these organelles in聽vivo and in a cell type specific fashion. We had previously shown that in oligodendrocytes and Schwann cells numerous peroxisomes reside in the cytoplasmic channels of 鈥渘on-compacted鈥?myelin. These organelles are smaller and biochemically distinct from non-myelin peroxisomes. Targeting peroxisomal functions in various cell types of the brain has demonstrated that oligodendroglial peroxisomes are specifically important for long-term integrity of the CNS. To visualize myelin peroxisomes in intact cells and tissues by live imaging, we have generated a novel line of transgenic mice for the expression of fluorescently tagged peroxisomes specifically in myelinating glia. This was achieved by modifying the gene for a photoconvertible mEos2 with a peroxisomal targeting signal type 1 (PTS1) and generating a fusion gene with the myelin-specific Cnp1 promoter. In the brain of resulting transgenic mice, peroxisomes are selectively labeled in oligodendrocytes. In this novel genetic tool, photoconversion of single peroxisomes from green to red fluorescence can be used to monitor the fate of single organelles and to determine the dynamics of PTS1-mediated protein import in the context of myelin diseases that affect peroxisomal functions.