In vivo bioluminescence reporter gene imaging for the activation of neuronal differentiation induced by the neuronal activator neurogenin 1 (Ngn1) in neuronal precursor cells

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• 作者：Hyun Jeong Oh ; Do Won Hwang ; Hyewon Youn…
• 关键词：Activation of neuronal differentiation ; In vivo optical imaging ; Neurogenin 1 ; Neuron ; specific reporter
• 刊名：European Journal of Nuclear Medicine and Molecular Imaging
• 出版年：2013
• 出版时间：October 2013
• 年：2013
• 卷：40
• 期：10
• 页码：1607-1617
• 全文大小：756KB
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• 作者单位：Hyun Jeong Oh (1) (5)
  Do Won Hwang (1) (2)
  Hyewon Youn (1) (3) (4)
  Dong Soo Lee (1) (5)
  
  1. Department of Nuclear Medicine, College of Medicine, Seoul National University, 101 Daehak-ro, Jongno-gu, Seoul, 110-744, Korea
  5. Department of Molecular Medicine and Biopharmaceutical Science, WCU Graduate School of Convergence Science and Technology, Seoul National University, Seoul, Korea
  2. Institute of Radiation Medicine, Medical Research Center, Seoul National University, Seoul, Korea
  3. Cancer Imaging Center, Seoul National University Cancer Hospital, Seoul National University, #207-4, Samsung Cancer Research Bldg. 103 Daehak-ro, Jongno-gu, 110-799, Seoul, Korea
  4. Laboratory of Molecular Imaging & Therapy, Cancer Research Institute, Seoul National University, Seoul, Korea
  
• ISSN：1619-7089

文摘

Purpose Facilitation of the ability of neuronal lineages derived from transplanted stem cells to differentiate is essential to improve the low efficacy of neuronal differentiation in stem cell therapy in vivo. Neurogenin 1 (Ngn1), a basic helix-loop-helix factor, has been used as an activator of neuronal differentiation. In this study, we monitored the in vivo activation of neuronal differentiation by Ngn1 in neuronal precursor cells using neuron-specific promoter-based optical reporters. Methods The NeuroD promoter coupled with the firefly luciferase reporter system (pNeuroD-Fluc) was used to monitor differentiation in F11 neuronal precursor cells. In vitro luciferase activity was measured and normalized by protein content. The in vivo-jetPEITM system was used for in vivo transgene delivery. The IVIS 100 imaging system was used to monitor in vivo luciferase activity. Results The Ngn1-induced neuronal differentiation of F11 cells generated neurite outgrowth within 2?days of Ngn1 induction. Immunofluorescence staining demonstrated that early and late neuronal marker expression (βIII-tubulin, NeuroD, MAP2, NF-M, and NeuN) was significantly increased at 3?days after treatment with Ngn1. When Ngn1 and the pNeuroD-Fluc vector were cotransfected into F11 cells, we observed an approximately 11-fold increase in the luciferase signal. An in vivo study showed that bioluminescence signals were gradually increased in Ngn1-treated F11 cells for up to 3?days. Conclusion In this study, we examined the in vivo tracking of neuronal differentiation induced by Ngn1 using an optical reporter system. This reporter system could be used effectively to monitor the activation efficiency of neuronal differentiation in grafted stem cells treated with Ngn1 for stem cell therapy.