Induction of Cell Cycle Arrest and Morphological Differentiation by Nurr1 and Retinoids in Dopamine MN9D Cells*

Dopamine cells are generated in the ventral midbrain during embryonic development. The progressive degeneration of these cells in patients with Parkinson's disease, and the potential therapeutic benefit by transplantation of in vitrogenerated dopamine cells, has triggered intense interest in understanding the process whereby these cells develop. Nurr1 is an orphan nuclear receptor essential for the development of midbrain dopaminergic neurons. However, the mechanism by which Nurr1 promotes dopamine cell differentiation has remained unknown. In this study we have used a dopamine-synthesizing cell line (MN9D) with immature characteristics to analyze the function of Nurr1 in dopamine cell development. The results demonstrate that Nurr1 can induce cell cycle arrest and a highly differentiated cell morphology in these cells. These two functions were both mediated through a DNA binding-dependent mechanism that did not require Nurr1 interaction with the heterodimerization partner retinoid X receptor. However, retinoids can promote the differentiation of MN9D cells independently of Nurr1. Importantly, the closely related orphan receptors NGFI-B and Nor1 were also able to induce cell cycle arrest and differentiation. Thus, the growth inhibitory activities of the NGFI-B/Nurr1/Nor1 orphan receptors, along with their widespread expression patterns both during development and in the adult, suggest a more general role in control of cell proliferation in the developing embryo and in adult tissues.

[1]  J. I. Izpisúa Belmonte,et al.  Evidence that Shh cooperates with a retinoic acid inducible co-factor to establish ZPA-like activity. , 1996, Development.

[2]  P. O’Farrell,et al.  Active repression of transcription by the engrailed homeodomain protein. , 1991, The EMBO journal.

[3]  F. Gage,et al.  Nurr1, an orphan nuclear receptor, is a transcriptional activator of endogenous tyrosine hydroxylase in neural progenitor cells derived from the adult brain. , 1999, Development.

[4]  J. Milbrandt,et al.  The orphan receptors NGFI-B and steroidogenic factor 1 establish monomer binding as a third paradigm of nuclear receptor-DNA interaction. , 1993, Molecular and cellular biology.

[5]  J. Smith,et al.  Inhibition of Xbra transcription activation causes defects in mesodermal patterning and reveals autoregulation of Xbra in dorsal mesoderm. , 1996, Development.

[6]  G. Duester.  Families of retinoid dehydrogenases regulating vitamin A function: production of visual pigment and retinoic acid. , 2000, European journal of biochemistry.

[7]  L. Olson,et al.  Fate of mesencephalic AHD2-expressing dopamine progenitor cells in NURR1 mutant mice. , 1999, Experimental cell research.

[8]  K. Umesono,et al.  Unique response pathways are established by allosteric interactions among nuclear hormone receptors , 1995, Cell.

[9]  G. Bruscalupi,et al.  Characterization of the response of growth and differentiation to lipoproteins and agents affecting cholesterol metabolism in murine neuroblastoma cells , 1994, International Journal of Developmental Neuroscience.

[10]  Marten P. Smidt,et al.  A second independent pathway for development of mesencephalic dopaminergic neurons requires Lmx1b , 2000, Nature Neuroscience.

[11]  K. Umesono,et al.  Functional inhibition of retinoic acid response by dominant negative retinoic acid receptor mutants. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

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

[13]  P. Chambon,et al.  Synergistic activation of retinoic acid (RA)-responsive genes and induction of embryonal carcinoma cell differentiation by an RA receptor alpha (RAR alpha)-, RAR beta-, or RAR gamma-selective ligand in combination with a retinoid X receptor-specific ligand , 1995, Molecular and cellular biology.

[14]  J. Milbrandt,et al.  Domains regulating transcriptional activity of the inducible orphan receptor NGFI-B. , 1992, The Journal of biological chemistry.

[15]  T. Perlmann,et al.  Activity of the Nurr1 Carboxyl-terminal Domain Depends on Cell Type and Integrity of the Activation Function 2* , 1999, The Journal of Biological Chemistry.

[16]  T. Perlmann,et al.  A novel pathway for vitamin A signaling mediated by RXR heterodimerization with NGFI-B and NURR1. , 1995, Genes & development.

[17]  K. Umesono,et al.  Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D3 receptors , 1991, Cell.

[18]  S. Minucci,et al.  RAR and RXR selective ligands cooperatively induce apoptosis and neuronal differentiation in P19 embryonal carcinoma cells , 1996, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[19]  S. Tetradis,et al.  Regulation of the nuclear orphan receptor Nur77 in bone by parathyroid hormone. , 2001, Biochemical and biophysical research communications.

[20]  F. Sharp,et al.  Stress induces zinc finger immediate early genes in the rat adrenal gland , 2000, Brain Research.

[21]  Miguel Beato,et al.  Steroid hormone receptors: Many Actors in search of a plot , 1995, Cell.

[22]  A. Fox,et al.  Immortalization of embryonic mesencephalic dopaminergic neurons by somatic cell fusion , 1991, Brain Research.

[23]  L. Olson,et al.  Cellular expression of the immediate early transcription factors Nurr1 and NGFI-B suggests a gene regulatory role in several brain regions including the nigrostriatal dopamine system. , 1996, Brain research. Molecular brain research.

[24]  I B Dawid,et al.  Retinoid X receptor (RXR) within the RXR-retinoic acid receptor heterodimer binds its ligand and enhances retinoid-dependent gene expression , 1997, Molecular and cellular biology.

[25]  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.

[26]  M. Baudry,et al.  Nurr1 mRNA expression in neonatal and adult rat brain following kainic acid-induced seizure activity. , 1998, Brain research. Molecular brain research.

[27]  A. Heller,et al.  Glial-derived neurotrophic factor (GDNF) induced morphological differentiation of an immortalized monoclonal hybrid dopaminergic cell line of mesencephalic neuronal origin , 1996, Brain Research.

[28]  Philippe Kastner,et al.  Nonsteroid nuclear receptors: What Are genetic studies telling us about their role in real life? , 1995, Cell.

[29]  F. Zindy,et al.  Postnatal neuronal proliferation in mice lacking Ink4d and Kip1 inhibitors of cyclin-dependent kinases. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[30]  J. Drouin,et al.  Novel dimeric Nur77 signaling mechanism in endocrine and lymphoid cells , 1997, Molecular and cellular biology.

[31]  R. Evans,et al.  Nuclear receptor that identifies a novel retinoic acid response pathway , 1990, Nature.

[32]  J. Manley,et al.  Functional domains of the Drosophila Engrailed protein. , 1993, The EMBO journal.

[33]  S. Bandoh,et al.  The NGFI-B subfamily of the nuclear receptor superfamily (review). , 1998, International journal of oncology.

[34]  Jacek Ostrowski,et al.  Two distinct actions of retinoid-receptor ligands , 1996, Nature.

[35]  J. Lehmann,et al.  Retinoids selective for retinoid X receptor response pathways. , 1992, Science.

[36]  S. Mcconnell,et al.  Constructing the cerebral cortex: Neurogenesis and fate determination , 1995, Neuron.

[37]  K. Yoshikawa.  Cell cycle regulators in neural stem cells and postmitotic neurons , 2000, Neuroscience Research.

[38]  T. Perlmann,et al.  Retinoic Acid Receptor/Retinoid X Receptor Heterodimers Can Be Activated through Both Subunits Providing a Basis for Synergistic Transactivation and Cellular Differentiation* , 1997, The Journal of Biological Chemistry.

[39]  T. Perlmann,et al.  Docosahexaenoic acid, a ligand for the retinoid X receptor in mouse brain. , 2000, Science.

[40]  R. Post,et al.  Rat nurr1 is prominently expressed in perirhinal cortex, and differentially induced in the hippocampal dentate gyrus by electroconvulsive vs. kindled seizures. , 1997, Brain research. Molecular brain research.

[41]  B. Bresnihan,et al.  Involvement of the nuclear orphan receptor NURR1 in the regulation of corticotropin-releasing hormone expression and actions in human inflammatory arthritis. , 2001, Arthritis and rheumatism.

[42]  N. Ohkura,et al.  Molecular cloning of a novel thyroid/steroid receptor superfamily gene from cultured rat neuronal cells. , 1994, Biochemical and biophysical research communications.

[43]  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.

[44]  J. Milbrandt,et al.  Participation of non-zinc finger residues in DNA binding by two nuclear orphan receptors. , 1992, Science.

[45]  C. Glass,et al.  Regulation of retinoid signalling by receptor polarity and allosteric control of ligand binding , 1994, Nature.

[46]  T. Iwawaki,et al.  Identification of a potential nurr1 response element that activates the tyrosine hydroxylase gene promoter in cultured cells. , 2000, Biochemical and biophysical research communications.

[47]  G Wolterink,et al.  A homeodomain gene Ptx3 has highly restricted brain expression in mesencephalic dopaminergic neurons. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[48]  J. Milbrandt.  Nerve growth factor induces a gene homologous to the glucocorticoid receptor gene , 1988, Neuron.

[49]  F. Sharp,et al.  Prolonged expression of zinc finger immediate‐early gene mRNAs and decreased protein synthesis following kainic acid induced seizures , 1999, The European journal of neuroscience.

[50]  R. Evans,et al.  The RXR heterodimers and orphan receptors , 1995, Cell.

[51]  J. Drouin,et al.  Heterodimerization between Members of the Nur Subfamily of Orphan Nuclear Receptors as a Novel Mechanism for Gene Activation , 1999, Molecular and Cellular Biology.

[52]  P. Chambon,et al.  A mutation mimicking ligand‐induced conformational change yields a constitutive RXR that senses allosteric effects in heterodimers , 1997, The EMBO journal.

[53]  U. Dräger,et al.  High levels of a retinoic acid-generating dehydrogenase in the meso-telencephalic dopamine system. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[54]  J. B. Jaynes,et al.  Inserting the Ftz homeodomain into engrailed creates a dominant transcriptional repressor that specifically turns off Ftz target genes in vivo. , 1995, Development.