Authors

William J Freed, Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Department of Health and Human Services (DHHS)
Jia Chen, Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Department of Health and Human Services (DHHS)
Cristina M Bäckman, Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Department of Health and Human Services (DHHS)
Catherine M Schwartz, Laboratory of Neurosciences, Intramural Research Program (IRP), National Institute on Aging, National Institutes of Health (NIH), Department of Health and Human Services (DHHS)
Tandis Vazin, Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Department of Health and Human Services (DHHS)
Jingli Cai, Farber Institute for Neurosciences, Thomas Jefferson UniversityFollow
Charles E Spivak, Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Department of Health and Human Services (DHHS)
Carl R Lupica, Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Department of Health and Human Services (DHHS)
Mahendra S Rao, Laboratory of Neurosciences, Intramural Research Program (IRP), National Institute on Aging, National Institutes of Health (NIH), Department of Health and Human Services (DHHS)
Xianmin Zeng, Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Department of Health and Human Services (DHHS)

Document Type

Article

Publication Date

1-9-2008

Comments

This article has been peer reviewed and is published in PLoS One 2008, 3(1): e1422. The published version is available at DOI: 10.1371/journal.pone.0001422. © Public Library of Science

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.

METHODOLOGY: 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.

PRINCIPAL FINDINGS: 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.

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