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


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2007-09-21

AUTHORS

Do Won Hwang, Joo Hyun Kang, Jae Min Jeong, June-Key Chung, Myung Chul Lee, Soonhag Kim, Dong Soo Lee

ABSTRACT

PurposeWe imaged neuronal differentiation in vivo using dual reporters (sodium iodide symporter [NIS] and luciferase) coupled to a neuron-specific enolase (NSE) promoter.MethodsPC12 (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 99mTc-pertechnetate gamma camera and bioluminescence imaging, respectively.ResultspNSE-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.ConclusionNSE 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. More... »

PAGES

135-145

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00259-007-0561-8

DOI

http://dx.doi.org/10.1007/s00259-007-0561-8

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1024725391

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/17885755


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