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

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European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

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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Acknowledgments

We thank Dr. Sanjiv Gambhir of Stanford University, USA, for kindly donating the TSTA vector construct. This work was supported by the Nano Bio Regenomics Project and Radiation Science Project of the Korean Science and Engineering Foundation.

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Authors and Affiliations

  1. Programs in Neuroscience, Seoul National University, Seoul, South Korea

    Do Won Hwang & Dong Soo Lee

  2. Department of Nuclear Medicine, Seoul National University College of Medicine, 28 Yongon-Dong, Jongno-Gu, Seoul, 110-744, South Korea

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

  3. Medical Research Center, Seoul National University College of Medicine, Seoul, South Korea

    Soonhag Kim

  4. Laboratory of Molecular Imaging and Therapy of Cancer Research Institute, Seoul National University College of Medicine, Seoul, South Korea

    Do Won Hwang, Jae Min Jeong & June-Key Chung

  5. Department of Nuclear Medicine, Seoul National University Hospital, 28 Yongon-Dong, Jongno-Gu, Seoul, 110-744, South Korea

    Dong Soo Lee

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  1. Do Won Hwang
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Correspondence to Soonhag Kim or Dong Soo Lee.

Additional information

Soonhag Kim and Dong Soo Lee contributed equally to this investigation.

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Hwang, D.W., Kang, J.H., Jeong, J.M. et al. Noninvasive in vivo monitoring of neuronal differentiation using reporter driven by a neuronal promoter. Eur J Nucl Med Mol Imaging 35, 135–145 (2008). https://doi.org/10.1007/s00259-007-0561-8

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  • DOI: https://doi.org/10.1007/s00259-007-0561-8

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