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The Journal of Neuroscience, January 15, 1999, 19(2):759-774

Identification and Characterization of Early Glial Progenitors Using a Transgenic Selection Strategy

Karen J. Chandross¹, Rick I. Cohen¹, Peter Paras Jr¹, Michel Gravel², Peter E. Braun², and Lynn D. Hudson¹

¹ National Institutes of Health, National Institute for Neurological Disorders and Stroke, Laboratory of Developmental Neurogenetics, Bethesda, Maryland 20892, and ² McGill University, Department of Biochemistry, Montreal, Quebec H3G1Y6, Canada

To define the spatiotemporal development of and simultaneously select for oligodendrocytes (OLs) and Schwann cells (SCs), transgenic mice were generated that expressed a bacterial -galactosidase (-gal) and neomycin phosphotransferase fusion protein (geo) under the control of murine 2'3'-cyclic nucleotide 3'-phosphodiesterase (muCNP) promoters I and II. Transgenic -gal activity was detected at embryonic day 12.5 in the ventral region of the rhombencephalon and spinal cord and in the neural crest. When cells from the rhombencephalon were cultured in the presence of G418, surviving cells differentiated into OLs, indicating that during development this brain region provides one source of OL progenitors. Postnatally, robust -gal activity was localized to OLs throughout the brain and was absent from astrocytes, neurons, and microglia or monocytes. In the sciatic nerve -gal activity was localized exclusively to SCs. Cultures from postnatal day 10 brain or sciatic nerve were grown in the presence of G418, and within 8-9 d exposure to antibiotic, 99% of all surviving cells were -gal-positive OLs or SCs. These studies demonstrate that the muCNP-geo transgenic mice are useful for identifying OLs and SCs beginning at early stages of the glial cell lineage and throughout their development. This novel approach definitively establishes that the -gal-positive cells identified in vivo are glial progenitors, as defined by their ability to survive antibiotic selection and differentiate into OLs or SCs in vitro. Moreover, this experimental paradigm facilitates the rapid and efficient selection of pure populations of mouse OLs and SCs and further underscores the use of cell-specific promoters in the purification of distinct cell types.

Key words: 2',3'-cyclic nucleotide 3'-phosphodiesterase; -galactosidase; CNS; development; glia; neomycin resistance; peripheral nervous system; selection; tissue culture

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Copyright © 1999 Society for Neuroscience  0270-6474/99/192759-16$05.00/0

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