Original Research Report In Vitro Generation of Dopaminergic Neurons from Adult Subventricular Zone Neural Progenitor Cells

The adult subventricular zone (SVZ) supports a population of cells that display the hallmarks of stem cells: they are self-renewing and multipotent—capable of generating neurons, oligodendrocytes, and astrocytes. In vivo, these adult neural stem cells (aNSCs) are fated primarily for a � -amino butyric acid (GABA)-ergic lineage of olfactory bulb interneurons, a small subpopulation of which is dopaminergic. Here, we investigate the plasticity of aNSCs in vitro, in particular, their ability to generate a specific neuronal lineage, midbrain dopamine neurons. Previous work using mouse embryonic stem (ES) cells showed that introduction of early developmental inductive cues, sonic hedgehog (SHH) and fibroblast growth factor-8 (FGF-8), directed ES cell-derived neuroepithelial cells to generate midbrain dopaminergic neurons, those lost in Parkinson’s disease. Placing aNSCs under similar culture conditions, immunocytochemistry and RT-PCR analysis revealed early dopaminergic neuron specification. However, aNSC-derived neurons remained morphologically immature, exhibiting concurrent nestin and tyrosine hydroxylase (TH) expression, with cell death occurring in the final differentiation stage. High-performance liquid chromatography (HPLC) analysis revealed that while aNSC-derived neurons released dopamine, release was not significantly increased following depolarization with K � . In

[1]  M. Chopp,et al.  The Sonic Hedgehog Pathway Mediates Carbamylated Erythropoietin-enhanced Proliferation and Differentiation of Adult Neural Progenitor Cells* , 2007, Journal of Biological Chemistry.

[2]  J. Partanen,et al.  Fibroblast Growth Factor Receptors Cooperate to Regulate Neural Progenitor Properties in the Developing Midbrain and Hindbrain , 2007, The Journal of Neuroscience.

[3]  Arturo Alvarez-Buylla,et al.  Mosaic Organization of Neural Stem Cells in the Adult Brain , 2007, Science.

[4]  M. Ekker,et al.  A Subpopulation of Olfactory Bulb GABAergic Interneurons Is Derived from Emx1- and Dlx5/6-Expressing Progenitors , 2007, The Journal of Neuroscience.

[5]  A. Prochiantz,et al.  Progressive Loss of Dopaminergic Neurons in the Ventral Midbrain of Adult Mice Heterozygote for Engrailed1 , 2007, The Journal of Neuroscience.

[6]  H. Okano,et al.  Subventricular Zone-Derived Neuroblasts Migrate and Differentiate into Mature Neurons in the Post-Stroke Adult Striatum , 2006, The Journal of Neuroscience.

[7]  Q. Deng,et al.  Identification of Intrinsic Determinants of Midbrain Dopamine Neurons , 2006, Cell.

[8]  A. Joyner,et al.  In vivo analysis of quiescent adult neural stem cells responding to Sonic hedgehog , 2005, Nature.

[9]  M. Götz,et al.  Neuronal fate determinants of adult olfactory bulb neurogenesis , 2005, Nature Neuroscience.

[10]  R. Pearce,et al.  Directed Differentiation of Dopaminergic Neuronal Subtypes from Human Embryonic Stem Cells , 2005, Stem cells.

[11]  Kwang-Soo Kim,et al.  The homeodomain transcription factor Pitx3 facilitates differentiation of mouse embryonic stem cells into AHD2-expressing dopaminergic neurons , 2005, Molecular and Cellular Neuroscience.

[12]  Alan Carleton,et al.  Sonic hedgehog controls stem cell behavior in the postnatal and adult brain , 2005, Development.

[13]  Ole Isacson,et al.  Intrastriatal Transforming Growth Factor α Delivery to a Model of Parkinson's Disease Induces Proliferation and Migration of Endogenous Adult Neural Progenitor Cells without Differentiation into Dopaminergic Neurons , 2004, The Journal of Neuroscience.

[14]  V. Tabar,et al.  Derivation of midbrain dopamine neurons from human embryonic stem cells. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[15]  M. Smidt,et al.  Homeobox gene Pitx3 and its role in the development of dopamine neurons of the substantia nigra , 2004, Cell and Tissue Research.

[16]  M. Picciotto,et al.  Nestin promoter/enhancer directs transgene expression to precursors of adult generated periglomerular neurons , 2004, The Journal of comparative neurology.

[17]  Theo Hagg,et al.  Dopaminergic nigrostriatal projections regulate neural precursor proliferation in the adult mouse subventricular zone , 2004, The European journal of neuroscience.

[18]  E. Wagner,et al.  Catecholamine Synthesizing Enzymes and Their Modulation by Immobilization Stress in Knockout Mice , 2004, Annals of the New York Academy of Sciences.

[19]  A. Álvarez-Buylla,et al.  For the Long Run Maintaining Germinal Niches in the Adult Brain , 2004, Neuron.

[20]  M. Li,et al.  Generation of embryonic stem cells and transgenic mice expressing green fluorescence protein in midbrain dopaminergic neurons , 2004, The European journal of neuroscience.

[21]  V. Palma,et al.  Hedgehog-GLI signaling regulates the behavior of cells with stem cell properties in the developing neocortex , 2003, Development.

[22]  M. Beal,et al.  Neural subtype specification of fertilization and nuclear transfer embryonic stem cells and application in parkinsonian mice , 2003, Nature Biotechnology.

[23]  W. Dauer,et al.  Parkinson's Disease Mechanisms and Models , 2003, Neuron.

[24]  Michael Dragunow,et al.  Increased cell proliferation and neurogenesis in the adult human Huntington's disease brain , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[25]  H. Baker,et al.  Dlx‐1 and Dlx‐2 expression in the adult mouse brain: Relationship to dopaminergic phenotypic regulation , 2003, The Journal of comparative neurology.

[26]  Wolfgang Wurst,et al.  The isthmic organizer signal FGF8 is required for cell survival in the prospective midbrain and cerebellum , 2003, Development.

[27]  J. García-Verdugo,et al.  Postnatal development of radial glia and the ventricular zone (VZ): a continuum of the neural stem cell compartment. , 2003, Cerebral cortex.

[28]  S. Weiss,et al.  Glycoprotein 130 Signaling Regulates Notch1Expression and Activation in the Self-Renewal of Mammalian Forebrain Neural Stem Cells , 2003, The Journal of Neuroscience.

[29]  M. Depew,et al.  DLX5 Regulates Development of Peripheral and Central Components of the Olfactory System , 2003, The Journal of Neuroscience.

[30]  Arturo Alvarez-Buylla,et al.  EGF Converts Transit-Amplifying Neurogenic Precursors in the Adult Brain into Multipotent Stem Cells , 2002, Neuron.

[31]  P. Hantraye,et al.  Intrastriatal sonic hedgehog injection increases Patched transcript levels in the adult rat subventricular zone , 2002, The European journal of neuroscience.

[32]  Jürgen Winkler,et al.  Long‐term survival and cell death of newly generated neurons in the adult rat olfactory bulb , 2002, The European journal of neuroscience.

[33]  O. Lindvall,et al.  Neuronal replacement from endogenous precursors in the adult brain after stroke , 2002, Nature Medicine.

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

[35]  R. McKay,et al.  Dopamine neurons derived from embryonic stem cells function in an animal model of Parkinson's disease , 2002, Nature.

[36]  H. Rohrer,et al.  Specification of catecholaminergic and serotonergic neurons , 2002, Nature Reviews Neuroscience.

[37]  Arturo Alvarez-Buylla,et al.  Neurogenesis in Adult Subventricular Zone , 2002, The Journal of Neuroscience.

[38]  H. Chun,et al.  Phenotypic Differentiation during Migration of Dopaminergic Progenitor Cells to the Olfactory Bulb , 2001, The Journal of Neuroscience.

[39]  Sally Temple,et al.  The development of neural stem cells , 2001, Nature.

[40]  Wolfgang Wurst,et al.  Fate of Midbrain Dopaminergic Neurons Controlled by the Engrailed Genes , 2001, The Journal of Neuroscience.

[41]  A. Sidow,et al.  A double-deletion mutation in the Pitx3 gene causes arrested lens development in aphakia mice. , 2001, Genomics.

[42]  Nadine Kabbani,et al.  Enhanced Proliferation, Survival, and Dopaminergic Differentiation of CNS Precursors in Lowered Oxygen , 2000, The Journal of Neuroscience.

[43]  K. Mizuseki,et al.  Induction of Midbrain Dopaminergic Neurons from ES Cells by Stromal Cell–Derived Inducing Activity , 2000, Neuron.

[44]  M. Pompeiano,et al.  Decreased apoptosis in proliferative and postmitotic regions of the caspase 3‐deficient embryonic central nervous system , 2000, The Journal of comparative neurology.

[45]  R. McKay,et al.  Efficient generation of midbrain and hindbrain neurons from mouse embryonic stem cells , 2000, Nature Biotechnology.

[46]  F. Gage,et al.  Mammalian neural stem cells. , 2000, Science.

[47]  J. García-Verdugo,et al.  Adult‐derived neural precursors transplanted into multiple regions in the adult brain , 1999, Annals of neurology.

[48]  E. Snyder,et al.  Induction of a midbrain dopaminergic phenotype in Nurr1-overexpressing neural stem cells by type 1 astrocytes , 1999, Nature Biotechnology.

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

[50]  D. van der Kooy,et al.  Adult Mammalian Forebrain Ependymal and Subependymal Cells Demonstrate Proliferative Potential, but only Subependymal Cells Have Neural Stem Cell Characteristics , 1999, The Journal of Neuroscience.

[51]  E. Parati,et al.  Epidermal and Fibroblast Growth Factors Behave as Mitogenic Regulators for a Single Multipotent Stem Cell-Like Population from the Subventricular Region of the Adult Mouse Forebrain , 1999, The Journal of Neuroscience.

[52]  A. Rosenthal,et al.  Specification of dopaminergic and serotonergic neurons in the vertebrate CNS , 1999, Current Opinion in Neurobiology.

[53]  G. Fishell,et al.  Telencephalic progenitors maintain anteroposterior identities cell autonomously , 1998, Current Biology.

[54]  M. Luskin.  Neuroblasts of the postnatal mammalian forebrain: their phenotype and fate. , 1998, Journal of neurobiology.

[55]  R. McKay,et al.  Transplantation of expanded mesencephalic precursors leads to recovery in parkinsonian rats , 1998, Nature Neuroscience.

[56]  M. Brand,et al.  Fgf8 is mutated in zebrafish acerebellar (ace) mutants and is required for maintenance of midbrain-hindbrain boundary development and somitogenesis. , 1998, Development.

[57]  J. Weiner,et al.  Programmed cell death is a universal feature of embryonic and postnatal neuroproliferative regions throughout the central nervous system , 1998, The Journal of comparative neurology.

[58]  J. Rubenstein,et al.  FGF and Shh Signals Control Dopaminergic and Serotonergic Cell Fate in the Anterior Neural Plate , 1998, Cell.

[59]  M. Palkovits,et al.  Dopamine Biosynthesis Is Selectively Abolished in Substantia Nigra/Ventral Tegmental Area but Not in Hypothalamic Neurons in Mice with Targeted Disruption of the Nurr1 Gene , 1998, Molecular and Cellular Neuroscience.

[60]  M. Smidt,et al.  Nurr1 is essential for the induction of the dopaminergic phenotype and the survival of ventral mesencephalic late dopaminergic precursor neurons. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[61]  T. Kosaka,et al.  How simple is the organization of the olfactory glomerulus?: the heterogeneity of so-called periglomerular cells , 1998, Neuroscience Research.

[62]  G. Martin,et al.  An Fgf8 mutant allelic series generated by Cre- and Flp-mediated recombination , 1998, Nature Genetics.

[63]  B J Hoffer,et al.  Dopamine neuron agenesis in Nurr1-deficient mice. , 1997, Science.

[64]  J. Parnavelas,et al.  Apoptosis and Its Relation to the Cell Cycle in the Developing Cerebral Cortex , 1997, The Journal of Neuroscience.

[65]  A. Álvarez-Buylla,et al.  Network of tangential pathways for neuronal migration in adult mammalian brain. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[66]  Andrew Lumsden,et al.  Patterning the Vertebrate Neuraxis , 1996, Science.

[67]  R. Bakay,et al.  Dopaminergic and gabaergic interneurons of the olfactory bulb are derived from the neonatal subventricular zone , 1996, International Journal of Developmental Neuroscience.

[68]  F. Gage,et al.  Differentiation of adult hippocampus-derived progenitors into olfactory neurons in vivo , 1996, Nature.

[69]  A. Blaschke,et al.  Widespread programmed cell death in proliferative and postmitotic regions of the fetal cerebral cortex. , 1996, Development.

[70]  Arturo Alvarez-Buylla,et al.  Chain Migration of Neuronal Precursors , 1996, Science.

[71]  J. Rubenstein,et al.  Longitudinal organization of the anterior neural plate and neural tube. , 1995, Development.

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

[73]  A. Joyner,et al.  Multiple developmental defects in Engrailed-1 mutant mice: an early mid-hindbrain deletion and patterning defects in forelimbs and sternum. , 1994, Development.

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

[75]  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.

[76]  S. Weiss,et al.  Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. , 1992, Science.

[77]  E G Jones,et al.  Evidence for coexistence of GABA and dopamine in neurons of the rat olfactory bulb , 1987, The Journal of comparative neurology.

[78]  E. Parati,et al.  Neural stem cells: an overview. , 2004, Journal of endocrinological investigation.

[79]  M. Ekker,et al.  Neural stem cell lineages are regionally specified, but not committed, within distinct compartments of the developing brain. , 2002, Development.

[80]  A. Farbman,et al.  The cell biology of olfaction , 1992 .