Sonic Hedgehog and FGF8: Inadequate Signals for the Differentiation of a Dopamine Phenotype in Mouse and Human Neurons in Culture
Embryonic mouse striatal neurons and human neurons derived from the NT2/hNT stem cell line can be induced, in culture, to express the dopaminergic (DA) biosynthetic enzyme tyrosine hydroxylase (TH). The novel expression of TH in these cells is signaled by the synergistic interaction of factors prese...
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Published in | Experimental neurology Vol. 169; no. 1; pp. 36 - 43 |
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Main Authors | , |
Format | Journal Article |
Language | English |
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Amsterdam
Elsevier Inc
01.05.2001
Elsevier |
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Abstract | Embryonic mouse striatal neurons and human neurons derived from the NT2/hNT stem cell line can be induced, in culture, to express the dopaminergic (DA) biosynthetic enzyme tyrosine hydroxylase (TH). The novel expression of TH in these cells is signaled by the synergistic interaction of factors present in the media, such as fibroblast growth factor 1 (FGF1) and one of several possible coactivators [DA, phorbol 12-myristate 13-acetate (TPA), isobutylmethylxanthine (IBMX), or forskolin]. Similarly, in vivo, it has recently been reported that the expression of TH in the developing midbrain is mediated by the synergy of FGF8 and the patterning molecule sonic hedgehog (Shh). In the present study, we examined whether the putative in vivo DA differentiation factors can similarly signal TH in our in vitro cell systems. We found that FGF8 and Shh induced TH expression in fewer than 2% of NT2/hNT cells and less than 5% of striatal neurons. The latter could be amplified to as much as 30% by increasing the concentration of growth factor 10-fold or by the addition of other competent coactivators (IBMX/forskolin, TPA, and DA). Additivity/inhibitor experiments indicated that FGF8 worked through traditional tyrosine kinase-initiated MAP/MEK signaling pathways. However, the Shh signal transduction cascade remained unclear. These data suggest that cues effective in vivo may be less successful in promoting the differentiation of a DA phenotype in mouse and human neurons in culture. Thus, our ability to generate DA neurons from different cell lines, for use in the treatment of Parkinson's disease, will depend on the identification of appropriate differentiation signals for each cell type under investigation. |
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AbstractList | Embryonic mouse striatal neurons and human neurons derived from the NT2/hNT stem cell line can be induced, in culture, to express the dopaminergic (DA) biosynthetic enzyme tyrosine hydroxylase (TH). The novel expression of TH in these cells is signaled by the synergistic interaction of factors present in the media, such as fibroblast growth factor 1 (FGF1) and one of several possible coactivators [DA, phorbol 12-myristate 13-acetate (TPA), isobutylmethylxanthine (IBMX), or forskolin]. Similarly, in vivo, it has recently been reported that the expression of TH in the developing midbrain is mediated by the synergy of FGF8 and the patterning molecule sonic hedgehog (Shh). In the present study, we examined whether the putative in vivo DA differentiation factors can similarly signal TH in our in vitro cell systems. We found that FGF8 and Shh induced TH expression in fewer than 2% of NT2/hNT cells and less than 5% of striatal neurons. The latter could be amplified to as much as 30% by increasing the concentration of growth factor 10-fold or by the addition of other competent coactivators (IBMX/forskolin, TPA, and DA). Additivity/inhibitor experiments indicated that FGF8 worked through traditional tyrosine kinase-initiated MAP/MEK signaling pathways. However, the Shh signal transduction cascade remained unclear. These data suggest that cues effective in vivo may be less successful in promoting the differentiation of a DA phenotype in mouse and human neurons in culture. Thus, our ability to generate DA neurons from different cell lines, for use in the treatment of Parkinson's disease, will depend on the identification of appropriate differentiation signals for each cell type under investigation. |
Author | Stull, Natalie D. Iacovitti, Lorraine |
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Keywords | stem cells differentiation fibroblast growth factor Parkinson's disease striatum tissue culture sonic hedgehog NT2 cells dopamine tyrosine hydroxylase Cell culture Stem cell Parkinson disease Tyrosine 3-monooxygenase Signal transduction Degenerative disease Protein-tyrosine kinase Human Tyrosine Nervous system diseases Dopamine Enzyme Transferases Rodentia Cell differentiation Gene expression Cerebral disorder Vertebrata Chemotherapy Phenotype Mammalia Treatment Mouse Animal Central nervous system disease Dopaminergic neuron Oxidoreductases Gene therapy Extrapyramidal syndrome |
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SubjectTerms | 1-Methyl-3-isobutylxanthine - pharmacology Animals Biological and medical sciences Biotechnology Cell Differentiation - drug effects Cell Differentiation - physiology Cells, Cultured Colforsin - pharmacology Corpus Striatum - cytology Corpus Striatum - embryology Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases differentiation dopamine Dopamine - metabolism Dopamine - pharmacology Dose-Response Relationship, Drug Drug Synergism Enzyme Inhibitors - pharmacology fibroblast growth factor Fibroblast Growth Factor 8 Fibroblast Growth Factors - pharmacology Fundamental and applied biological sciences. Psychology Gene therapy Health. Pharmaceutical industry Hedgehog Proteins Humans Industrial applications and implications. Economical aspects MAP Kinase Signaling System - drug effects Medical sciences Mice Neurology Neurons - cytology Neurons - drug effects Neurons - metabolism NT2 cells Parkinson's disease Phenotype Proteins - pharmacology Signal Transduction - drug effects Signal Transduction - physiology sonic hedgehog stem cells striatum tissue culture Tissue Plasminogen Activator - pharmacology Trans-Activators Tyrosine 3-Monooxygenase - biosynthesis tyrosine hydroxylase |
Title | Sonic Hedgehog and FGF8: Inadequate Signals for the Differentiation of a Dopamine Phenotype in Mouse and Human Neurons in Culture |
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