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PMID: 18183302 Published · epublish English Journal Article Research Support, N.I.H., Extramural

Gene expression profile of neuronal progenitor cells derived from hESCs: activation of chromosome 11p15.5 and comparison to human dopaminergic neurons.

PloS one ·Vol. 3 ·No. 1 ·2008-01-09 ·Pages e1422

Freed WJ, Chen J, Bäckman CM, Schwartz CM, Vazin T, Cai J, Spivak CE, Lupica CR, Rao MS, Zeng X

Abstract

We initiated differentiation of human embryonic stem cells (hESCs) into dopamine neurons, obtained a purified population of neuronal precursor cells by cell sorting, and determined patterns of gene transcription. Dopaminergic differentiation of hESCs was initiated by culturing hESCs with a feeder layer of PA6 cells. Differentiating cells were then sorted to obtain a pure population of PSA-NCAM-expressing neuronal precursors, which were then analyzed for gene expression using Massive Parallel Signature Sequencing (MPSS). Individual genes as well as regions of the genome which were activated were determined. A number of genes known to be involved in the specification of dopaminergic neurons, including MSX1, CDKN1C, Pitx1 and Pitx2, as well as several novel genes not previously associated with dopaminergic differentiation, were expressed. Notably, we found that a specific region of the genome located on chromosome 11p15.5 was highly activated. This region contains several genes which have previously been associated with the function of dopaminergic neurons, including the gene for tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis, IGF2, and CDKN1C, which cooperates with Nurr1 in directing the differentiation of dopaminergic neurons. Other genes in this region not previously recognized as being involved in the functions of dopaminergic neurons were also activated, including H19, TSSC4, and HBG2. IGF2 and CDKN1C were also found to be highly expressed in mature human TH-positive dopamine neurons isolated from human brain samples by laser capture. The present data suggest that the H19-IGF2 imprinting region on chromosome 11p15.5 is involved in the process through which undifferentiated cells are specified to become neuronal precursors and/or dopaminergic neurons.

MeSH Terms
Cell Differentiation/genetics Cell Line Cell Separation Chromosomes, Human, Pair 11 Dopamine/metabolism Embryonic Stem Cells/cytology Flow Cytometry Gene Expression Profiling Humans Immunohistochemistry Neurons/cytology,metabolism Polymerase Chain Reaction
Chemicals
Dopamine
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Freed William J
Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Baltimore, Maryland, USA. [email protected]
Chen Jia
Bäckman Cristina M
Schwartz Catherine M
Vazin Tandis
Cai Jingli
Spivak Charles E
Lupica Carl R
Rao Mahendra S
Zeng Xianmin
References (66)
66 references, click to expand
  1. Development of dopaminergic neurons in the mammalian brain.
    Cell Mol Life Sci. 2006 Jan;63(2):187-206 PMID: 16389456
  2. Identification of intrinsic determinants of midbrain dopamine neurons.
    Cell. 2006 Jan 27;124(2):393-405 PMID: 16439212
  3. Imprinting in clusters: lessons from Beckwith-Wiedemann syndrome.
    Trends Genet. 1997 Aug;13(8):330-4 PMID: 9260520
  4. Isolation of lineage-restricted neuronal precursors from multipotent neuroepithelial stem cells.
    Neuron. 1997 Oct;19(4):773-85 PMID: 9354325
  5. In vitro cloning of complex mixtures of DNA on microbeads: physical separation of differentially expressed cDNAs.
    Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1665-70 PMID: 10677516
  6. Gene expression analysis by massively parallel signature sequencing (MPSS) on microbead arrays.
    Nat Biotechnol. 2000 Jun;18(6):630-4 PMID: 10835600
  7. Loss of imprinted genes and paternal SUR1 mutations lead to focal form of congenital hyperinsulinism.
    Horm Res. 2000;53 Suppl 1:2-6 PMID: 10895035
  8. Sequence conservation and variability of imprinting in the Beckwith-Wiedemann syndrome gene cluster in human and mouse.
    Hum Mol Genet. 2000 Jul 22;9(12):1829-41 PMID: 10915772
  9. Induction of midbrain dopaminergic neurons from ES cells by stromal cell-derived inducing activity.
    Neuron. 2000 Oct;28(1):31-40 PMID: 11086981
  10. Ablation of NF1 function in neurons induces abnormal development of cerebral cortex and reactive gliosis in the brain.
    Genes Dev. 2001 Apr 1;15(7):859-76 PMID: 11297510
  11. Proliferation of progenitor cells in the adult rat brain correlates with the presence of vimentin-expressing astrocytes.
    Glia. 2001 Jun;34(4):253-66 PMID: 11360298
  12. Multilineage differentiation from human embryonic stem cell lines.
    Stem Cells. 2001;19(3):193-204 PMID: 11359944
  13. Altered sensitivity of CD81-deficient mice to neurobehavioral effects of cocaine.
    Brain Res Mol Brain Res. 2001 May 20;90(1):68-74 PMID: 11376857
  14. Unbalanced expression of 11p15 imprinted genes in focal forms of congenital hyperinsulinism: association with a reduction to homozygosity of a mutation in ABCC8 or KCNJ11.
    Am J Pathol. 2001 Jun;158(6):2177-84 PMID: 11395395
  15. A murine dopamine neuron-specific cDNA library and microarray: increased COX1 expression during methamphetamine neurotoxicity.
    Neurobiol Dis. 2001 Oct;8(5):822-33 PMID: 11592851
  16. Generation of dopaminergic neurons and pigmented epithelia from primate ES cells by stromal cell-derived inducing activity.
    Proc Natl Acad Sci U S A. 2002 Feb 5;99(3):1580-5 PMID: 11818560
  17. 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
  18. 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
  19. BMP4 initiates human embryonic stem cell differentiation to trophoblast.
    Nat Biotechnol. 2002 Dec;20(12):1261-4 PMID: 12426580
  20. The Caenorhabditis elegans dopaminergic system: opportunities for insights into dopamine transport and neurodegeneration.
    Annu Rev Pharmacol Toxicol. 2003;43:521-44 PMID: 12415122
  21. Complex history of a chromosomal paralogy region: insights from amphioxus aromatic amino acid hydroxylase genes and insulin-related genes.
    Mol Biol Evol. 1998 Nov;15(11):1373-80 PMID: 12572601
  22. Stable expression of hrGFP by mouse embryonic stem cells: promoter activity in the undifferentiated state and during dopaminergic neural differentiation.
    Stem Cells. 2003;21(6):647-53 PMID: 14595124
  23. Spatiotemporal patterns of gene expression during fetal monkey brain development.
    Brain Res Dev Brain Res. 2003 Dec 19;146(1-2):99-106 PMID: 14643016
  24. p57(Kip2) cooperates with Nurr1 in developing dopamine cells.
    Proc Natl Acad Sci U S A. 2003 Dec 23;100(26):15619-24 PMID: 14671317
  25. Making connections: the development of mesencephalic dopaminergic neurons.
    Brain Res Dev Brain Res. 2003 Dec 30;147(1-2):3-21 PMID: 14741747
  26. Spontaneous differentiation of germ cells from human embryonic stem cells in vitro.
    Hum Mol Genet. 2004 Apr 1;13(7):727-39 PMID: 14962983
  27. CD81-induced behavioural changes during chronic cocaine administration: in vivo gene delivery with regulatable lentivirus.
    Eur J Neurosci. 2004 Mar;19(6):1621-33 PMID: 15066158
  28. Properties of pluripotent human embryonic stem cells BG01 and BG02.
    Stem Cells. 2004;22(3):292-312 PMID: 15153607
  29. Derivation of midbrain dopamine neurons from human embryonic stem cells.
    Proc Natl Acad Sci U S A. 2004 Aug 24;101(34):12543-8 PMID: 15310843
  30. Generation of tyrosine hydroxylase positive neurons from human embryonic stem cells after coculture with cellular substrates and exposure to GDNF.
    Stem Cells. 2004;22(5):669-74 PMID: 15342931
  31. The structure and expression of a novel gene activated in early mouse embryogenesis.
    EMBO J. 1988 Mar;7(3):673-81 PMID: 3396539
  32. Identification of antisense transcripts of the chicken insulin-like growth factor-II gene.
    J Mol Endocrinol. 1991 Oct;7(2):145-54 PMID: 1930626
  33. Embryonic stem cell lines derived from human blastocysts.
    Science. 1998 Nov 6;282(5391):1145-7 PMID: 9804556
  34. Two novel genes in the center of the 11p15 imprinted domain escape genomic imprinting.
    Hum Mol Genet. 1999 Apr;8(4):683-90 PMID: 10072438
  35. SWiP-1: novel SOCS box containing WD-protein regulated by signalling centres and by Shh during development.
    Mech Dev. 1999 Apr;82(1-2):79-94 PMID: 10354473
  36. Genomic imprinting of IGF-II and H19 in adult human pancreatic tissues.
    Digestion. 1999 Sep-Oct;60(5):477-83 PMID: 10473973
  37. Dopaminergic differentiation of human embryonic stem cells.
    Stem Cells. 2004;22(6):925-40 PMID: 15536184
  38. In vivo gene delivery of urokinase-type plasminogen activator with regulatable lentivirus induces behavioural changes in chronic cocaine administration.
    Eur J Neurosci. 2004 Dec;20(12):3473-88 PMID: 15610180
  39. Structural and functional analysis of a 0.5-Mb chicken region orthologous to the imprinted mammalian Ascl2/Mash2-Igf2-H19 region.
    Genome Res. 2005 Jan;15(1):154-65 PMID: 15590938
  40. How imprinting centres work.
    Cytogenet Genome Res. 2006;113(1-4):81-9 PMID: 16575166
  41. The H19 gene: regulation and function of a non-coding RNA.
    Cytogenet Genome Res. 2006;113(1-4):188-93 PMID: 16575179
  42. Cognitive dysfunction in NF1 knock-out mice may result from altered vesicular trafficking of APP/DRD3 complex.
    BMC Neurosci. 2006;7:22 PMID: 16524466
  43. Excitability of dopamine neurons: modulation and physiological consequences.
    CNS Neurol Disord Drug Targets. 2006 Feb;5(1):79-97 PMID: 16613555
  44. CTCF mediates interchromosomal colocalization between Igf2/H19 and Wsb1/Nf1.
    Science. 2006 Apr 14;312(5771):269-72 PMID: 16614224
  45. Identification of novel genes regulated in the developing human ventral mesencephalon.
    Exp Neurol. 2006 Apr;198(2):427-37 PMID: 16473350
  46. Transcriptome coexpression map of human embryonic stem cells.
    BMC Genomics. 2006;7:103 PMID: 16670017
  47. The imprinted H19 noncoding RNA is a primary microRNA precursor.
    RNA. 2007 Mar;13(3):313-6 PMID: 17237358
  48. Sox-2 is expressed by glial and progenitor cells and Pax-6 is expressed by neuroblasts in the human subventricular zone.
    Exp Neurol. 2007 Apr;204(2):828-31 PMID: 17291498
  49. IGF and FGF cooperatively establish the regulatory stem cell niche of pluripotent human cells in vitro.
    Nature. 2007 Aug 30;448(7157):1015-21 PMID: 17625568
  50. Parental imprinting of the human H19 gene.
    FEBS Lett. 1992 Aug 31;309(1):25-8 PMID: 1380925
  51. In vitro differentiation of E-N-CAM expressing rat neural precursor cells isolated by FACS during prenatal development.
    J Neurosci Res. 1994 Feb 15;37(3):359-73 PMID: 8176758
  52. Expression of the imprinted gene H19 in the human fetus.
    Mol Reprod Dev. 1994 Jul;38(3):239-46 PMID: 7917273
  53. Lack of imprinting of the human dopamine D4 receptor (DRD4) gene.
    Am J Med Genet. 1996 Apr 9;67(2):229-31 PMID: 8723054
  54. Altered cell differentiation and proliferation in mice lacking p57KIP2 indicates a role in Beckwith-Wiedemann syndrome.
    Nature. 1997 May 8;387(6629):151-8 PMID: 9144284
  55. DNA methylation and imprinting: why bother?
    Trends Genet. 1997 Aug;13(8):323-9 PMID: 9260519
  56. Karyotypic stability, genotyping, differentiation, feeder-free maintenance, and gene expression sampling in three human embryonic stem cell lines derived prior to August 9, 2001.
    Stem Cells Dev. 2004 Dec;13(6):585-97 PMID: 15684826
  57. Monitoring early differentiation events in human embryonic stem cells by massively parallel signature sequencing and expressed sequence tag scan.
    Stem Cells Dev. 2004 Dec;13(6):694-715 PMID: 15684837
  58. The homeodomain transcription factor Pitx3 facilitates differentiation of mouse embryonic stem cells into AHD2-expressing dopaminergic neurons.
    Mol Cell Neurosci. 2005 Feb;28(2):241-52 PMID: 15691706
  59. Directed differentiation of dopaminergic neuronal subtypes from human embryonic stem cells.
    Stem Cells. 2005 Jun-Jul;23(6):781-90 PMID: 15917474
  60. The Hedgehog-inducible ubiquitin ligase subunit WSB-1 modulates thyroid hormone activation and PTHrP secretion in the developing growth plate.
    Nat Cell Biol. 2005 Jul;7(7):698-705 PMID: 15965468
  61. An atlas of human gene expression from massively parallel signature sequencing (MPSS).
    Genome Res. 2005 Jul;15(7):1007-14 PMID: 15998913
  62. De-differentiation-derived mesenchymal stem cells demonstrate selective repression in H19 bioregulatory RNA gene expression.
    Differentiation. 2005 Jul;73(6):294-302 PMID: 16138830
  63. Dopaminergic neurones: much more than dopamine?
    Br J Pharmacol. 2005 Sep;146(2):167-9 PMID: 16025140
  64. The role of chromatin structure in regulating the expression of clustered genes.
    Nat Rev Genet. 2005 Oct;6(10):775-81 PMID: 16160692
  65. NTera2: a model system to study dopaminergic differentiation of human embryonic stem cells.
    Stem Cells Dev. 2005 Oct;14(5):517-34 PMID: 16305337
  66. Development of the mesencephalic dopaminergic neuron system is compromised in the absence of neurogenin 2.
    Development. 2006 Feb;133(3):507-16 PMID: 16396906
Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0001422
Email
[email protected]
Published
2008-01-09
Epub
2008-00-09
Pages
e1422
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC2170519
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pone.0001422 PMC Full Text View journal details
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