Early Postnatal Astroglial Cells Produce Multilineage Precursors and Neural Stem Cells In Vivo

To identify the fates that astroglial cells can attain in the postnatal brain, we generated mice carrying an inducible Cre recombinase (Cre-ERT2) controlled by the human GFAP promoter (hGFAP). In mice carrying the GCE (hGFAP-Cre-ERT2) transgene, OHT (4-hydroxy-tamoxifen) injections induced Cre recombination in astroglial cells at postnatal day 5 and allowed us to permanently tag these cells with reporter genes. Three days after recombination, reporter-tagged cells were quiescent astroglial cells that expressed the stem cell marker LeX in the subventricular zone (SVZ) and dentate gyrus (DG). After 2–4 weeks, the tagged GFAP lineage included proliferating progenitors expressing the neuronal marker Dcx (Doublecortin) in the SVZ and the DG. After 4 weeks, the GFAP lineage generated mature neurons in the olfactory bulb (OB), DG, and, strikingly, also in the cerebral cortex. A major portion of all neurons in the DG and OB born at the end of the first postnatal week were generated from GFAP+ cells. In addition to neurons, mature oligodendrocytes and astrocytes populating the cerebral cortex and white matter were also the progeny of GFAP+ astroglial ancestors. Thus, genetic fate mapping of postnatal GFAP+ cells reveals that they seed the postnatal brain with neural progenitors/stem cells that in turn give rise to neural precursors and their mature neuronal and oligodendrocytic progeny in many CNS regions, including the cerebral cortex.

[1]  D. Steindler,et al.  Identification of a multipotent astrocytic stem cell in the immature and adult mouse brain. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Daniel A. Lim,et al.  Subventricular Zone Astrocytes Are Neural Stem Cells in the Adult Mammalian Brain , 1999, Cell.

[3]  K. McCarthy,et al.  GFAP-positive progenitor cells produce neurons and oligodendrocytes throughout the CNS , 2006, Molecular and Cellular Neuroscience.

[4]  Philippe Soriano.  Generalized lacZ expression with the ROSA26 Cre reporter strain , 1999, Nature Genetics.

[5]  B. Sauer.  Inducible gene targeting in mice using the Cre/lox system. , 1998, Methods.

[6]  J. Partanen,et al.  Fibroblast Growth Factor Receptor 1 Is Required for the Proliferation of Hippocampal Progenitor Cells and for Hippocampal Growth in Mouse , 2022 .

[7]  M. Götz,et al.  Isolation of radial glial cells by fluorescent-activated cell sorting reveals a neuronal lineage. , 2000, Development.

[8]  J. Goldman,et al.  Gliogenic and neurogenic progenitors of the subventricular zone: Who are they, where did they come from, and where are they going? , 2003, Glia.

[9]  D. van der Kooy,et al.  The ablation of glial fibrillary acidic protein‐positive cells from the adult central nervous system results in the loss of forebrain neural stem cells but not retinal stem cells , 2003, The European journal of neuroscience.

[10]  M. Greenberg,et al.  Regulation of gliogenesis in the central nervous system by the JAK-STAT signaling pathway. , 1997, Science.

[11]  M. Sofroniew,et al.  GFAP-expressing progenitors are the principal source of constitutive neurogenesis in adult mouse forebrain , 2004, Nature Neuroscience.

[12]  C. Lois,et al.  Proliferating subventricular zone cells in the adult mammalian forebrain can differentiate into neurons and glia. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Brenner,et al.  GFAP Transgenic Mice , 1996, Methods.

[14]  Pasko Rakic,et al.  A golgi study of radial glial cells in developing monkey telencephalon: Morphogenesis and transformation into astrocytes , 1979, Anatomy and Embryology.

[15]  Brent A. Reynolds,et al.  Neural stem cells in the adult mammalian forebrain: A relatively quiescent subpopulation of subependymal cells , 1994, Neuron.

[16]  V S Caviness,et al.  Identification of radial glial cells within the developing murine central nervous system: studies based upon a new immunohistochemical marker. , 1988, Brain research. Developmental brain research.

[17]  P. Bartlett,et al.  The Adult Mouse Hippocampal Progenitor Is Neurogenic But Not a Stem Cell , 2005, The Journal of Neuroscience.

[18]  J. de Vellis,et al.  Transition between immature radial glia and mature astrocytes studied with a monoclonal antibody to vimentin. , 1984, Brain research.

[19]  S. Weiss,et al.  The Ciliary Neurotrophic Factor/Leukemia Inhibitory Factor/gp130 Receptor Complex Operates in the Maintenance of Mammalian Forebrain Neural Stem Cells , 2001, The Journal of Neuroscience.

[20]  G. Fishell,et al.  Radial Glia Serve as Neuronal Progenitors in All Regions of the Central Nervous System , 2004, Neuron.

[21]  V. Caviness,et al.  Dynamic changes in the density of radial glial fibers of the developing murine cerebral wall: A quantitative immunohistological analysis , 1992, The Journal of comparative neurology.

[22]  A. F. Schinder,et al.  Neuronal Differentiation in the Adult Hippocampus Recapitulates Embryonic Development , 2005, The Journal of Neuroscience.

[23]  C. Englund,et al.  Pax6, Tbr2, and Tbr1 Are Expressed Sequentially by Radial Glia, Intermediate Progenitor Cells, and Postmitotic Neurons in Developing Neocortex , 2005, The Journal of Neuroscience.

[24]  F. Vaccarino,et al.  Chronic hypoxia up-regulates fibroblast growth factor ligands in the perinatal brain and induces fibroblast growth factor-responsive radial glial cells in the sub-ependymal zone , 2002, Neuroscience.

[25]  P. Chambon,et al.  Conditional site-specific recombination in mammalian cells using a ligand-dependent chimeric Cre recombinase. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[26]  P. Rakic,et al.  Cell Proliferation Without Neurogenesis in Adult Primate Neocortex , 2001, Science.

[27]  Maria B. Luskin,et al.  Restricted proliferation and migration of postnatally generated neurons derived from the forebrain subventricular zone , 1993, Neuron.

[28]  M. Edwards,et al.  Organization of radial glia and related cells in the developing murine CNS. An analysis based upon a new monoclonal antibody marker , 1990, Neuroscience.

[29]  E. Geisert,et al.  Myelin/oligodendrocyte‐specific protein: A novel surface membrane protein that associates with microtubules , 1991, Journal of neuroscience research.

[30]  J. García-Verdugo,et al.  Astrocytes Give Rise to New Neurons in the Adult Mammalian Hippocampus , 2001, The Journal of Neuroscience.

[31]  T. Weissman,et al.  Neurons derived from radial glial cells establish radial units in neocortex , 2001, Nature.

[32]  D. van der Kooy,et al.  Adult Rodent Neurogenic Regions: The Ventricular Subependyma Contains Neural Stem Cells, But the Dentate Gyrus Contains Restricted Progenitors , 2002, The Journal of Neuroscience.

[33]  W. Wurst,et al.  Inducible gene deletion in astroglia and radial glia—A valuable tool for functional and lineage analysis , 2006, Glia.

[34]  F. Vaccarino,et al.  Fibroblast Growth Factor 2 Is Required for Maintaining the Neural Stem Cell Pool in the Mouse Brain Subventricular Zone , 2004, Developmental Neuroscience.

[35]  S. Weiss,et al.  Generation of Functional Radial Glial Cells by Embryonic and Adult Forebrain Neural Stem Cells , 2003, The Journal of Neuroscience.

[36]  M. Götz,et al.  Neuronal or Glial Progeny Regional Differences in Radial Glia Fate , 2003, Neuron.

[37]  S. Temple,et al.  LeX/ssea-1 Is Expressed by Adult Mouse CNS Stem Cells, Identifying Them as Nonependymal , 2002, Neuron.

[38]  P Chambon,et al.  Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ER(T) and Cre-ER(T2) recombinases. , 1999, Nucleic acids research.

[39]  V. Gallo,et al.  NG2-expressing cells in the subventricular zone are type C–like cells and contribute to interneuron generation in the postnatal hippocampus , 2004, The Journal of cell biology.

[40]  V. Gallo,et al.  Postnatal Neurogenesis and Gliogenesis in the Olfactory Bulb from NG2-Expressing Progenitors of the Subventricular Zone , 2004, The Journal of Neuroscience.

[41]  J. Goldman,et al.  Glial progenitors of the neonatal subventricular zone differentiate asynchronously, leading to spatial dispersion of glial clones and to the persistence of immature glia in the adult mammalian CNS. , 2004, Developmental biology.

[42]  A. Messing,et al.  GFAP promoter directs astrocyte-specific expression in transgenic mice , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[43]  P. Rakic,et al.  Molecular and Morphological Heterogeneity of Neural Precursors in the Mouse Neocortical Proliferative Zones , 2006, The Journal of Neuroscience.

[44]  Caiying Guo,et al.  Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon cre‐mediated excision , 2000, Genesis.

[45]  François Guillemot,et al.  Mash1 specifies neurons and oligodendrocytes in the postnatal brain , 2004, The EMBO journal.

[46]  N. Šestan,et al.  Midline radial glia translocation and corpus callosum formation require FGF signaling , 2006, Nature Neuroscience.

[47]  P Chambon,et al.  Ligand-activated site-specific recombination in mice. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[48]  Michaela Brenner.  Structure and Transcriptional Regulation of the GFAP Gene , 1994, Brain pathology.

[49]  J. García-Verdugo,et al.  Radial glia give rise to adult neural stem cells in the subventricular zone. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[50]  J. García-Verdugo,et al.  Cellular Composition and Three-Dimensional Organization of the Subventricular Germinal Zone in the Adult Mammalian Brain , 1997, The Journal of Neuroscience.

[51]  K. Miyazono,et al.  Synergistic signaling in fetal brain by STAT3-Smad1 complex bridged by p300. , 1999, Science.

[52]  Laura R. Ment,et al.  Cortical neurogenesis enhanced by chronic perinatal hypoxia , 2006, Experimental Neurology.

[53]  Philipp Berger,et al.  Tamoxifen-inducible glia-specific Cre mice for somatic mutagenesis in oligodendrocytes and Schwann cells , 2003, Molecular and Cellular Neuroscience.

[54]  A. Kriegstein,et al.  Radial glia diversity: A matter of cell fate , 2003, Glia.

[55]  D. Steindler,et al.  Editorial: To be glial or not—how glial are the precursors of neurons in development and adulthood? , 2003, Glia.

[56]  C. Gross,et al.  Neurogenesis in the neocortex of adult primates. , 1999, Science.

[57]  Z. Kaprielian,et al.  Oligodendrocyte and astrocyte development in rodents: An in situ and immunohistological analysis during embryonic development , 2002, Glia.