Elsevier

Developmental Biology

Volume 218, Issue 2, 15 February 2000, Pages 259-274
Developmental Biology

Regular Article
Purification and Analysis of in Vivo-Differentiated Oligodendrocytes Expressing the Green Fluorescent Protein

https://doi.org/10.1006/dbio.1999.9574Get rights and content
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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

GFP
differentiated oligodendrocyte
patch clamp
primary culture
live cell imaging

Cited by (0)

1

Present address: Department of Anatomy, Virginia Commonwealth University, Richmond, VA.

2

Present address: Department of Physiology and Neurobiology, University of Connecticut, Storrs, CT.