Abstract
We have constructed a recombinant adenovirus expression vector carrying the human neurotrophin-3 (NT-3) receptor TrkC (tyrosine protein kinase C) gene (rAd-TrkC; 2478 bp) and confirmed the expression of the encoded TrkC in green fluorescent protein (GFP)-murine neural stem cells (NSCs) by reverse transcription polymerase chain reaction (RT-PCR), Western blot analysis, and immunocytochemistry. The activity of the expressed rAd-TrkC was verified in vitro by evaluating dose-related responses of NSCs to NT-3, a TrkC specific ligand. TrkC-GFP-NSCs had a significantly higher percentage of neuronal differentiation when treated with NT-3 relative to the rAd-LacZ control cells (55.2% vs. 29.8%; P<0.05, chi(2) test). Thus, our rAd-TrkC vector can transfect NSCs and produce functional TrkC receptors to promote neuronal differentiation of NSCs.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Adenoviridae / genetics
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Animals
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Animals, Newborn
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Cell Culture Techniques
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Cell Differentiation / drug effects
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Cell Differentiation / genetics*
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Cells, Cultured
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Dose-Response Relationship, Drug
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Gene Expression Regulation / genetics
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Genetic Vectors / genetics*
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Mice
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Molecular Biology / methods
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Neurons / cytology
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Neurons / drug effects
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Neurons / metabolism*
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Neurotrophin 3 / metabolism
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Neurotrophin 3 / pharmacology
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Receptor, trkC / agonists
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Receptor, trkC / genetics*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Stem Cells / cytology
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Stem Cells / drug effects
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Stem Cells / metabolism*
Substances
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Neurotrophin 3
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Recombinant Fusion Proteins
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Green Fluorescent Proteins
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Receptor, trkC