title: Gene expression profile of neuronal progenitor cells derived from hESCs: activation of chromosome 11p15.5 and comparison to human dopaminergic neurons.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Baltimore, Maryland, USA. wfreed@mail.nih.gov
authors.forename: William J
authors.initials: WJ
authors.lastname: Freed
authors.forename: Jia
authors.initials: J
authors.lastname: Chen
authors.forename: Cristina M
authors.initials: CM
authors.lastname: Bäckman
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Schwartz
authors.forename: Tandis
authors.initials: T
authors.lastname: Vazin
authors.forename: Jingli
authors.initials: J
authors.lastname: Cai
authors.forename: Charles E
authors.initials: CE
authors.lastname: Spivak
authors.forename: Carl R
authors.initials: CR
authors.lastname: Lupica
authors.forename: Mahendra S
authors.initials: MS
authors.lastname: Rao
authors.forename: Xianmin
authors.initials: X
authors.lastname: Zeng
mesh_terms: D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D002469:Cell Separation; D002880:Chromosomes, Human, Pair 11; D004298:Dopamine / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D016133:Polymerase Chain Reaction
chemical_list: D004298:Dopamine