Changes of gene expression profiles during neuronal differentiation of central nervous system precursors treated with ascorbic acid
- Changes of gene expression profiles during neuronal differentiation of central nervous system precursors treated with ascorbic acid
- ascorbic acid; CNS precursor; cDNA mi-croarray; dopaminergic neurons
- Issue Date
- JOURNAL OF NEUROSCIENCE RESEARCH, v. 78, No. 1, Page. 29-37
- Ascorbic acid (AA) has been shown to increase the yield of dopaminergic (DA) neurons
derived from basic fibroblast growth factor (bFGF)-expanded mesencephalic precursors.
To understand the molecular mechanisms underlying this phenomenon, we used cDNA
microarray analysis to examine differential expression of neuronal genes following
AA treatment. The putative precursor cells were isolated from E13 rat ventral
mesencephalons and expanded in the presence of bFGF. Cells were incubated in
mitogen-free media supplemented with 200 M AA or were left untreated as a control,
and total RNA was isolated at different time points (expansion stage and 1, 3, and
6 days after induction of differentiation) and subjected to cDNA microarray analysis.
Differentiation was evaluated by Western blot analysis and immunocytochemistry of
neuron-specific markers. AA treatment of the mesencephalic precursors increased the
expression of neuronal (MAP2) and astrocytic (glial fibrillary acidic protein)
markers and the percentage of tyrosine hydroxylase (TH)-positive cells. The
microarray analysis revealed that 12 known genes were up-regulated and 20 known
genes were down-regulated in expansion-stage AA-treated cells. Six days after the
induction of differentiation, AA-treated cells showed up-regulation of 48 known
genes and down-regulation of 5 known genes. Our results identified several proteins, such as transferrin, S-100, and somatostatin, as being differentially regulated in AA-treated mesencephalic precursors. This novel result may lead to a better understanding of the molecular mechanisms underlying the AA-induced differentiation of mesencephalic precursors into DA neurons and may form the basis for improved DA neuronal production for treatment of Parkinson`s disease patients.
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