Home LiteratureArticle Details
PMID: 12453046 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Genetic engineering of mouse embryonic stem cells by Nurr1 enhances differentiation and maturation into dopaminergic neurons.

The European journal of neuroscience ·Vol. 16 ·No. 10 ·2002-11-00 ·Pages 1829-38

Chung S, Sonntag KC, Andersson T, Bjorklund LM, Park JJ, Kim DW, Kang UJ, Isacson O, Kim KS

Abstract

Nurr1 is a transcription factor critical for the development of midbrain dopaminergic (DA) neurons. This study modified mouse embryonic stem (ES) cells to constitutively express Nurr1 under the elongation factor-1alpha promoter. The Nurr1-expression in ES cells lead to up-regulation of all DA neuronal markers tested, resulting in about a 4- to 5-fold increase in the proportion of DA neurons. In contrast, other neuronal and glial markers were not significantly changed by Nurr1 expression. It was also observed that there was an additional 4-fold increase in the number of DA neurons in Nurr1-expressing clones following treatment with Shh, FGF8 and ascorbic acid. Several lines of evidence suggest that these neurons may represent midbrain DA neuronal phenotypes; firstly, they coexpress midbrain DA markers such as aromatic L-amino acid decarboxylase, calretinin, and dopamine transporter, in addition to tyrosine hydroxylase and secondly, they do not coexpress other neurotransmitters such as GABA or serotonin. Finally, consistent with an increased number of DA neurons, the Nurr1 transduction enhanced the ability of these neurons to produce and release DA in response to membrane depolarization. This study demonstrates an efficient genetic manipulation of ES cells that facilitates differentiation to midbrain DA neurons, and it will serve as a framework of genetic engineering of ES cells by key transcription factor to regulate their cell fate.

MeSH Terms
Animals Brain Stem/metabolism Cell Culture Techniques Cell Differentiation/genetics DNA-Binding Proteins/genetics,metabolism Dopamine/metabolism Genetic Engineering/methods Immunohistochemistry Mice Neurons/metabolism Nuclear Receptor Subfamily 4, Group A, Member 2 Plasmids Pluripotent Stem Cells/metabolism Reverse Transcriptase Polymerase Chain Reaction Serotonin/metabolism Transcription Factors/genetics,metabolism Transfection Tyrosine 3-Monooxygenase/metabolism
Chemicals
DNA-Binding Proteins Nr4a2 protein, mouse Nuclear Receptor Subfamily 4, Group A, Member 2 Transcription Factors Serotonin Tyrosine 3-Monooxygenase Dopamine
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Chung Sangmi
Udall Parkinson's Disease Research Center of Excellence, Neuroregeneration Laboratory, McLean Hospital/Harvard Medical School, Belmont, MA 02478, USA.
Sonntag Kai-C
Andersson Therese
Bjorklund Lars M
Park Jae-Joon
Kim Dong-Wook
Kang Un Jung
Isacson Ole
Kim Kwang-Soo
References (49)
49 references, click to expand
  1. Differential regulation of transcription by the NURR1/NUR77 subfamily of nuclear transcription factors.
    Gene Expr. 1996;5(3):169-79 PMID: 8882640
  2. 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.
    Brain Res Mol Brain Res. 1996 Sep 5;41(1-2):111-20 PMID: 8883941
  3. Development of neuronal precursor cells and functional postmitotic neurons from embryonic stem cells in vitro.
    Mech Dev. 1996 Sep;59(1):89-102 PMID: 8892235
  4. Single factors direct the differentiation of stem cells from the fetal and adult central nervous system.
    Genes Dev. 1996 Dec 15;10(24):3129-40 PMID: 8985182
  5. Dopamine neuron agenesis in Nurr1-deficient mice.
    Science. 1997 Apr 11;276(5310):248-50 PMID: 9092472
  6. Role of aromatic L-amino acid decarboxylase for dopamine replacement by genetically modified fibroblasts in a rat model of Parkinson's disease.
    J Neurochem. 1997 Nov;69(5):2055-63 PMID: 9349551
  7. A homeodomain gene Ptx3 has highly restricted brain expression in mesencephalic dopaminergic neurons.
    Proc Natl Acad Sci U S A. 1997 Nov 25;94(24):13305-10 PMID: 9371841
  8. Blastula-stage stem cells can differentiate into dopaminergic and serotonergic neurons after transplantation.
    Exp Neurol. 1998 Jan;149(1):28-41 PMID: 9454612
  9. Nurr1 is essential for the induction of the dopaminergic phenotype and the survival of ventral mesencephalic late dopaminergic precursor neurons.
    Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):4013-8 PMID: 9520484
  10. 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.
    Mol Cell Neurosci. 1998 May;11(1-2):36-46 PMID: 9608532
  11. FGF and Shh signals control dopaminergic and serotonergic cell fate in the anterior neural plate.
    Cell. 1998 May 29;93(5):755-66 PMID: 9630220
  12. Specification of dopaminergic and serotonergic neurons in the vertebrate CNS.
    Curr Opin Neurobiol. 1999 Feb;9(1):26-36 PMID: 10072377
  13. Transcriptional control of neurotransmitter phenotype.
    Curr Opin Neurobiol. 1999 Feb;9(1):47-53 PMID: 10072363
  14. Induction of a midbrain dopaminergic phenotype in Nurr1-overexpressing neural stem cells by type 1 astrocytes.
    Nat Biotechnol. 1999 Jul;17(7):653-9 PMID: 10404157
  15. Embryonic stem cell-derived glial precursors: a source of myelinating transplants.
    Science. 1999 Jul 30;285(5428):754-6 PMID: 10427001
  16. Nurr1, an orphan nuclear receptor, is a transcriptional activator of endogenous tyrosine hydroxylase in neural progenitor cells derived from the adult brain.
    Development. 1999 Sep;126(18):4017-26 PMID: 10457011
  17. Characterization of the 5'-flanking region of the human dopamine transporter gene.
    Brain Res Mol Brain Res. 1999 Dec 10;74(1-2):167-74 PMID: 10640687
  18. A selective group of dopaminergic neurons express Nurr1 in the adult mouse brain.
    Brain Res. 1999 Dec 18;851(1-2):125-32 PMID: 10642835
  19. Calretinin and calbindin-D28k in dopaminergic neurons of the rat midbrain: a triple-labeling immunohistochemical study.
    Brain Res. 1999 Oct 30;846(1):129-36 PMID: 10536220
  20. Fate of mesencephalic AHD2-expressing dopamine progenitor cells in NURR1 mutant mice.
    Exp Cell Res. 1999 Dec 15;253(2):737-46 PMID: 10585298
  21. Dopamine transporter gene expression in rat mesencephalic dopaminergic neurons is increased by direct interaction with target striatal cells in vitro.
    Brain Res Mol Brain Res. 1996 Jul;39(1-2):160-6 PMID: 8804724
  22. Cloning and expression of a glutamate transporter from mouse brain.
    Neurosci Lett. 1993 Sep 3;159(1-2):183-6 PMID: 7903437
  23. Target cells modulate dopamine transporter gene expression during brain development.
    Neuroreport. 1994 May 9;5(9):1145-8 PMID: 7521683
  24. Embryonic stem cells differentiated in vitro as a novel source of cells for transplantation.
    Cell Transplant. 1996 Mar-Apr;5(2):131-43 PMID: 8689027
  25. Derivation of completely cell culture-derived mice from early-passage embryonic stem cells.
    Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8424-8 PMID: 8378314
  26. Identification of a new brain-specific transcription factor, NURR1.
    Mol Endocrinol. 1992 Dec;6(12):2129-35 PMID: 1491694
  27. Embryonic stem cells alone are able to support fetal development in the mouse.
    Development. 1990 Nov;110(3):815-21 PMID: 2088722
  28. Efficient generation of midbrain and hindbrain neurons from mouse embryonic stem cells.
    Nat Biotechnol. 2000 Jun;18(6):675-9 PMID: 10835609
  29. A shift from reversible to irreversible X inactivation is triggered during ES cell differentiation.
    Mol Cell. 2000 Apr;5(4):695-705 PMID: 10882105
  30. Identification of a potential nurr1 response element that activates the tyrosine hydroxylase gene promoter in cultured cells.
    Biochem Biophys Res Commun. 2000 Aug 11;274(3):590-5 PMID: 10924322
  31. Embryonic stem cells go dopaminergic.
    Neuron. 2000 Oct;28(1):11-4 PMID: 11086977
  32. Induction of midbrain dopaminergic neurons from ES cells by stromal cell-derived inducing activity.
    Neuron. 2000 Oct;28(1):31-40 PMID: 11086981
  33. Nigrostriatal innervation is preserved in Nurr1-null mice, although dopaminergic neuron precursors are arrested from terminal differentiation.
    Brain Res Mol Brain Res. 2000 Dec 8;84(1-2):67-78 PMID: 11113533
  34. Embryoid bodies: an in vitro model of mouse embryogenesis.
    Exp Physiol. 2000 Nov;85(6):645-51 PMID: 11187960
  35. Growth factors regulate the survival and fate of cells derived from human neurospheres.
    Nat Biotechnol. 2001 May;19(5):475-9 PMID: 11329020
  36. Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets.
    Science. 2001 May 18;292(5520):1389-94 PMID: 11326082
  37. Differentiation of embryonic stem cell-derived dopaminergic neurons is enhanced by survival-promoting factors.
    Mech Dev. 2001 Jul;105(1-2):93-104 PMID: 11429285
  38. Stem cells with brainpower.
    Nat Biotechnol. 2001 Dec;19(12):1117-8 PMID: 11731775
  39. In vitro differentiation of transplantable neural precursors from human embryonic stem cells.
    Nat Biotechnol. 2001 Dec;19(12):1129-33 PMID: 11731781
  40. Neural progenitors from human embryonic stem cells.
    Nat Biotechnol. 2001 Dec;19(12):1134-40 PMID: 11731782
  41. Orphan nuclear receptor Nurr1 is essential for Ret expression in midbrain dopamine neurons and in the brain stem.
    Mol Cell Neurosci. 2001 Dec;18(6):649-63 PMID: 11749040
  42. Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model.
    Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):2344-9 PMID: 11782534
  43. Analysis of different promoter systems for efficient transgene expression in mouse embryonic stem cell lines.
    Stem Cells. 2002;20(2):139-45 PMID: 11897870
  44. Dopamine neurons derived from embryonic stem cells function in an animal model of Parkinson's disease.
    Nature. 2002 Jul 4;418(6893):50-6 PMID: 12077607
  45. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  46. Establishment in culture of pluripotential cells from mouse embryos.
    Nature. 1981 Jul 9;292(5819):154-6 PMID: 7242681
  47. Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells.
    Proc Natl Acad Sci U S A. 1981 Dec;78(12):7634-8 PMID: 6950406
  48. HPRT-deficient (Lesch-Nyhan) mouse embryos derived from germline colonization by cultured cells.
    Nature. 1987 Mar 19-25;326(6110):292-5 PMID: 3821905
  49. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells.
    Cell. 1987 Nov 6;51(3):503-12 PMID: 2822260
Article Info
Journal
The European journal of neuroscience
Abbr.
Eur J Neurosci
ISSN
0953-816X
DOI
10.1046/j.1460-9568.2002.02255.x
Published
2002-11-00
Pages
1829-38
Language
English
Region
France
NLM ID
8918110
PMCID
PMC2610444
Subset
IM
Grants
NIMH NIH HHS · R01 MH048866 · United States
NINDS NIH HHS · R01 NS032080 · United States
NINDS NIH HHS · NS32080 · United States
NIMH NIH HHS · R29 MH048866 · United States
NINDS NIH HHS · P50 NS039793-03S1 · United States
NIMH NIH HHS · MH48866 · United States
NINDS NIH HHS · P50 NS039793 · United States
NINDS NIH HHS · R01 NS084869 · United States
NINDS NIH HHS · P50NS39793 · United States
External Links
PubMed source DOI: 10.1046/j.1460-9568.2002.02255.x PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2