Gene Expression Profile of Neuronal Progenitor Cells Derived from hESCs: Activation of Chromosome 11p15.5 and Comparison to Human Dopaminergic Neurons

Abstract

BackgroundWe 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.MethodologyDopaminergic 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 FindingsA 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.ConclusionsThe 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.

Mesenchymal Stem Cells from Human Umbilical Cord Express Preferentially Secreted Factors Related to Neuroprotection, Neurogenesis, and Angiogenesis

“…In contrast, the principal overrepresented processes associated with BM-MSCs include those related to ossification and skeletal system development (Figure 1C). In WJ-MSCs, neurogenesis transcriptional factors such as SOX11 [38] and PITX1 [39], as well as transcriptional factors associated with angiogenesis such as FOXF1 [40,41], were highly expressed, and these findings were confirmed by RT-qPCR (Figure 1D). Receptors involved in neurogenesis and angiogenesis, including CDH2, NRP2 and FLT1 , were also more abundant in WJ-MSCs (Figure 1D).…”

Neuroplasticity, axonal guidance and micro‐RNA genes are associated with morphine self‐administration behavior

“…A gene of considerable interest in the VS network is H19 . This gene contains miRNA miR-675 which has been postulated to regulate dopaminergic neuron function (Freed et al, 2008) thus indicating a role of miRNAs in drug seeking behavior. The identification of H19 as an important filtered SA gene is particularly intriguing given our recent finding that H19 is negatively correlated with morphine tolerance (Tapocik et al, 2009).…”

“…H19 is intriguing since it was found negatively associated with our self-administration (present study) and tolerance data (Tapocik et al, 2009) in the VS and PAG, respectively. This primary miRNA gene gives rise to miR-675 in humans and mice (Cai and Cullen, 2007) and the H19 imprinting region is involved in dopamine neuron differentiation (Freed et al, 2008). Since H19 was found associated with tolerance and self-administration it may be advantageous to pharmacologically target this candidate in chronic drug administration settings.…”

Genome Wide Profiling of Dopaminergic Neurons Derived from Human Embryonic and Induced Pluripotent Stem Cells

“…Development and use of newer technologies can help circumvent this caveat. For example, we have previously used FACS to enrich PSA-NCAM-expressing neural precursors [30], and other researchers have used a floor plate-specific cell surface marker Corin to enrich for mouse dopaminergic neurons [31,32]. Nevertheless, neither of these methods results in isolation of pure population of mature dopaminergic neurons, and thus far cell surface markers that would enable sorting of dopaminergic neurons have yet to be discovered.…”