Browse

Noninvasive in vivo monitoring of neuronal differentiation using reporter driven by a neuronal promoter

Cited 32 time in Web of Science Cited 31 time in Scopus
Authors
Hwang, Do Won; Kang, Joo Hyun; Jeong, Jae Min; Chung, June-Key; Lee, Myung Chul; Kim, Soonhag; Lee, Dong Soo
Issue Date
2007-09-22
Publisher
Springer Verlag
Citation
Eur J Nucl Med Mol Imaging. 2008 Jan;35(1):135-45. Epub 2007 Sep 21.
Keywords
AnimalsCell Differentiation/*geneticsCell LineGene Expression RegulationGenes, Reporter/*geneticsGenetic VectorsHumansIodine RadioisotopesLuciferases/genetics/metabolismLuminescenceNeurons/*cytology/*metabolism/radionuclide imagingPhosphopyruvate Hydratase/genetics/metabolismPromoter Regions, Genetic/*geneticsRatsSymporters/genetics/metabolismTransfection
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://hdl.handle.net/10371/28346
DOI
https://doi.org/10.1007/s00259-007-0561-8
Files in This Item:
There are no files associated with this item.
Appears in Collections:
College of Medicine/School of Medicine (의과대학/대학원)Program in Cancer Biology (협동과정-종양생물학전공)Journal Papers (저널논문_협동과정-종양생물학전공)
  • mendeley

Items in S-Space are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse