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Purification and analysis of in vivo-differentiated oligodendrocytes expressing the green fluorescent protein

Item Type:Article
Title:Purification and analysis of in vivo-differentiated oligodendrocytes expressing the green fluorescent protein
Creators Name:Fuss, B. and Mallon, B. and Phan, T. and Ohlemeyer, C. and Kirchhoff, F. and Nishiyama, A. and Macklin, W.B.
Abstract:A complete understanding of the molecular mechanisms involved in the formation and repair of the central nervous system myelin sheath requires an unambiguous identification and isolation of in vivo-differentiated myelin- forming cells. In order to develop a novel tool for the analysis of in vivo- differentiated oligodendrocytes, we generated transgenic mice expressing a red-shifted variant of the green fluorescent protein under the control of the proteolipid protein promoter. We demonstrate here that green fluorescent protein-derived fluorescence in the central nervous system of 9-day- to 7- week-old mice is restricted to mature oligodendrocytes, as determined by its spatiotemporal appearance and by both immunocytochemical and electrophysiological criteria. Green fluorescent protein-positive oligodendrocytes could easily be visualized in live and fixed tissue. Furthermore, we show that this convenient and reliable identification now allows detailed physiological analyses of differentiated oligodendrocytes in situ. In addition, we developed a novel tissue culture system for in vivo- differentiated oligodendrocytes. Initial data using this system indicate that, for oligodendrocytes isolated after differentiation in vivo, as yet unidentified factors secreted by astrocytes are necessary for survival and/or reappearance of a mature phenotype in culture.
Keywords:Differentiated Oligodendrocyte, GFP, Live Cell Imaging, Patch Clamp, Primary Culture, Animals, Mice
Source:Developmental Biology
Publisher:Academic Press
Page Range:259-274
Date:15 February 2000
Official Publication:https://doi.org/10.1006/dbio.1999.9574
PubMed:View item in PubMed

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