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GFAP-expressing progenitors are the principal source of constitutive neurogenesis in adult mouse forebrain

Nature Neuroscience volume 7, pages 1233–1241 (2004)Cite this article

Abstract

Establishing the cellular identity in vivo of adult multipotent neural progenitors is fundamental to understanding their biology. We used two transgenic strategies to determine the relative contribution of glial fibrillary acidic protein (GFAP)-expressing progenitors to constitutive neurogenesis in the adult forebrain. Transgenically targeted ablation of dividing GFAP-expressing cells in the adult mouse subependymal and subgranular zones stopped the generation of immunohistochemically identified neuroblasts and new neurons in the olfactory bulb and the hippocampal dentate gyrus. Transgenically targeted cell fate mapping showed that essentially all neuroblasts and neurons newly generated in the adult mouse forebrain in vivo, and in adult multipotent neurospheres in vitro, derived from progenitors that expressed GFAP. Constitutively dividing GFAP-expressing progenitors showed predominantly bipolar or unipolar morphologies with significantly fewer processes than non-neurogenic multipolar astrocytes. These findings identify morphologically distinctive GFAP-expressing progenitor cells as the predominant sources of constitutive adult neurogenesis, and provide new methods for manipulating and investigating these cells.

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Figure 1: Expression of transgene-derived HSV-TK and GFAP and labeling by BrdU in the SEZ and SGZ of adult GFAP-TK mice.
Figure 2: Depletion of BrdU labeling and neuroblast staining by progressive ablation of dividing GFAP-TK cells in the SEZ and SGZ.
Figure 3: Chronic ablation of dividing GFAP-TK cells stops the generation of new neurons in the adult olfactory bulb (OB) and dentate gyrus (DG).
Figure 4: Cre/loxP-mediated fate mapping of GFAP-expressing progenitors.
Figure 5: Adult neuroblasts and adult-born neurons express reporter protein in GFAP-Cre reporter mice in vitro and in vivo.
Figure 6: Dividing, GFAP-expressing progenitors in the adult SEZ and SGZ have a bipolar or unipolar phenotype that differs from nondividing multipolar stellate astrocytes.

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Acknowledgements

This work was supported by grants from the US National Institutes of Health (NINDS) NS042693 and NS47386, and a Stein/Oppenheimer Award. ADRG is supported by a Ford Foundation predoctoral fellowship. We thank M.E. Sislak and T. Chiem for technical assistance. We thank F. Myhren and Clavis Pharma for eGCV, and P. Borgese and Hoffman La Roche for GCV.

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  1. Department of Neurobiology and Brain Research Institute, University of California, Los Angeles, 90095-1763, California, USA

    A Denise R Garcia, Ngan B Doan, Tetsuya Imura, Toby G Bush & Michael V Sofroniew

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Supplementary information

Supplementary Fig. 1

Subpopulations of neurons express reporter protein in the olfactory bulbs and dentate gyrus, but not in the striatum. Single channel (a1-a2, b1-b2, c1-c2) and merged images (a3, b3, c3) of the confocal micrographs seen in Figure 5h, i, and k, respectively, showing βGal (green, a1) or GFP (green, b1, c1) and NeuN (blue, a2, b2, c2) double immunofluorescence staining in the striatum (a1-a3), olfactory bulb (b1-b3), and dentate gyrus (c1-c3). (a1-a3) In the striatum, NeuN-positive neurons do not express reporter protein (βGal); a βGal-positive cell that does not co-label with NeuN has the appearance of an astrocyte. (b1-b3) In the granule cell layer (GCL) of olfactory bulb, a subset of NeuN-positive cells are double labeled with GFP (arrows). (c1-c3) In the GCL of the dentate gyrus, a subset of NeuN-positive cells are double labeled with GFP (arrows); a GFP-positive cell that does not co-label with NeuN has the appearance of an astrocyte (arrowhead). (PDF 153 kb)

Supplementary Fig. 2

Dividing GFAP-positive/TK-positive cells in the SEZ and SGZ have a bipolar or unipolar morphology. Single channel (a1-a3, b1-b3) and merged images (a4, b4) of the confocal micrographs seen in Figure 6a and b. Triple immunofluorescence staining for BrdU (red), GFAP (green) and TK (blue) identifies triple stained cells in the SEZ (a1-a4) and SGZ (b1-b4). Triple stained areas where red, green and blue overlap appear white in the overlay images. The cell in (a) exhibits triple staining in much of the cell body. The cell in (b) exhibits white triple staining only in a few regions where GFAP positive filaments extend into the cell body, however, examination of all three channels separately and in overlay shows that the cell is clearly triple labeled. (PDF 141 kb)

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Garcia, A., Doan, N., Imura, T. et al. GFAP-expressing progenitors are the principal source of constitutive neurogenesis in adult mouse forebrain. Nat Neurosci 7, 1233–1241 (2004). https://doi.org/10.1038/nn1340

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