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European Journal of Nuclear Medicine and Molecular Imaging

Erschienen in:

01.01.2008 | Original Article

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

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

Erschienen in: European Journal of Nuclear Medicine and Molecular Imaging | Ausgabe 1/2008

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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 ^99mTc-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.

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Titel: Noninvasive in vivo monitoring of neuronal differentiation using reporter driven by a neuronal promoter
verfasst von: Do Won Hwang
Joo Hyun Kang
Jae Min Jeong
June-Key Chung
Myung Chul Lee
Soonhag Kim
Dong Soo Lee
Publikationsdatum: 01.01.2008
Verlag: Springer-Verlag
Erschienen in: European Journal of Nuclear Medicine and Molecular Imaging / Ausgabe 1/2008
Print ISSN: 1619-7070
Elektronische ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-007-0561-8

Springer Medizin

Abstract

Purpose

Methods

Results

Conclusion

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