Recombinant adenovirus vector-mediated functional expression of neurotropin-3 receptor (TrkC) in neural stem cells.
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.
Wang
Jun-Mei
JM
Division of Neuroscience, Department of Histology and Embryology, Zhongshan Medical College, Sun Yat-sen University, Guangzhou, China.
Zeng
Yuan-Shan
YS
Liu
Ran-Yi
RY
Huang
Wen-Lin
WL
Xiong
Yi
Y
Wang
Yan-Hua
YH
Chen
Shui-Jun
SJ
Teng
Yang D
YD
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008967:Molecular Biology / Q000379:methods; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D020933:Neurotrophin 3 / Q000378:metabolism / Q000494:pharmacology; D020812:Receptor, trkC / Q000819:agonists / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
D020933:Neurotrophin 3; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D020812:Receptor, trkC