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Noninvasive in vivo monitoring of neuronal differentiation using reporter driven by a neuronal promoter
Cited 33 time in
Web of Science
Cited 34 time in Scopus
- Authors
- Issue Date
- 2007-09-22
- Publisher
- Springer Verlag
- Citation
- Eur J Nucl Med Mol Imaging. 2008;35(1):135-45
- Keywords
- Animals ; Cell Differentiation/*genetics ; Cell Line ; Gene Expression Regulation ; Genes, Reporter/*genetics ; Genetic Vectors ; Humans ; Iodine Radioisotopes ; Luciferases/genetics/metabolism ; Luminescence ; Neurons/*cytology/*metabolism/radionuclide imaging ; Phosphopyruvate Hydratase/genetics/metabolism ; Promoter Regions, Genetic/*genetics ; Rats ; Symporters/genetics/metabolism ; Transfection
- Abstract
- PURPOSE: We imaged neuronal differentiation in vivo using dual reporters (sodium iodide symporter [NIS] and luciferase) coupled to a neuron-specific enolase (NSE) promoter. METHODS: PC12 (NSE positive) and F11 cells were transfected with a bicistronic (NIS and luciferase; pNSE-NF) or a luciferase (pNSE-Fluc) reporter coupled to the NSE promoter. Weak NSE promoter activity was overcome by a two-step transcriptional amplification (TSTA) system (pNSE-TSTA-Fluc). In vivo, NIS and luciferase expression were examined using a (99m)Tc-pertechnetate gamma camera and bioluminescence imaging, respectively. RESULTS: pNSE-NF-transfected PC12 cells showed 3-fold higher radioiodine uptakes and >100-fold higher luciferase activity than parental cells. NIS or luciferase activity was not detected in pNSE-NF-transfected HeLa cells. When F11 cells were differentiated into neurons by db-cAMP, NIS and luciferase activities increased 4-fold compared to those without treatment, which was confirmed by Western blot and RT-PCR of NSE. In vivo in pNSE-NF-transfected F11 cells, db-cAMP treatment increased the luciferase activity but not the scintigraphic activity. In vitro, pNSE-TSTA-Fluc produced 130-fold higher luciferase activity than pNSE-Fluc and neuronal differentiation showed 4-fold higher activity from both pNSE-TSTA-Fluc and pNSE-Fluc than before differentiation. In vivo, in pNSE-TSTA-Fluc-transfected F11 cells, luciferase activity increased after neuronal differentiation. In vivo luciferase activity persisted up to 2 days after db-cAMP-induced neuronal differentiation. CONCLUSION: NSE promoter-driven dual reporter transgenes revealed the possibility of in vivo imaging of neuronal differentiation, which was further enabled by high amplification using a TSTA system. We propose that this strategy be used to follow the transplanted stem cells during differentiation in live animals.
- ISSN
- 1619-7089 (Electronic)
- Language
- English
- URI
- http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=17885755
http://www.springerlink.com/content/a00w277558086p0v/fulltext.pdf
https://hdl.handle.net/10371/67782
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