Bioluminescence imaging of stroke-induced endogenous neural stem cell response

Neurobiology of Disease

Sarah-Ann Aelvoet¹  Zeger Debyser²  Greetje Vande Velde³  Veerle Reumers⁴  Rik Gijsbers⁵  Uwe Himmelreich⁵  Chris Van den Haute⁶  Caroline Vandeputte⁷  Irina Thiry⁷  Sylvie De Swaef⁷  Mathias Hoehn⁷  Tracy D. Farr⁷  Jesus Pascual-Brazo⁷  Koen Van Laere⁸  Veerle Baekelandt⁸ 

[1] Division of Nuclear Medicine, University Hospital and KU Leuven, 3000 Leuven, Flanders, Belgium;KU Leuven, Biomedical MRI, Department of Imaging and Pathology, 3000 Leuven, Flanders, Belgium;KU Leuven, Leuven Viral Vector Core, 3000 Leuven, Flanders, Belgium;KU Leuven, Molecular Small Animal Imaging Center, MOSAIC, KU Leuven, 3000 Leuven, Flanders, Belgium;In-vivo-NMR Laboratory, Max Planck Institute for Neurological Research, 50931 Cologne, Germany;KU Leuven, Laboratory for Molecular Virology and Gene Therapy, Department of Pharmaceutical and Pharmacological Sciences, 3000 Leuven, Flanders, Belgium;KU Leuven, Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, 3000 Leuven, Flanders, Belgium;KU Leuven, Molecular Small Animal Imaging Center, MOSAIC, KU Leuven, 3000 Leuven, Flanders, Belgium;

关键词: Bioluminescence imaging;    Cre-Flex lentiviral vector;    Endogenous neural stem cells;    Nestin-Cre mice;    Stroke;   

DOI  :  

来源: DOAJ

【 摘 要 】

Brain injury following stroke affects neurogenesis in the adult mammalian brain. However, a complete understanding of the origin and fate of the endogenous neural stem cells (eNSCs) in vivo is missing. Tools and technology that allow non-invasive imaging and tracking of eNSCs in living animals will help to overcome this hurdle.In this study, we aimed to monitor eNSCs in a photothrombotic (PT) stroke model using in vivo bioluminescence imaging (BLI). In a first strategy, inducible transgenic mice expressing firefly luciferase (Fluc) in the eNSCs were generated. In animals that received stroke, an increased BLI signal originating from the infarct region was observed. However, due to histological limitations, the identity and exact origin of cells contributing to the increased BLI signal could not be revealed. To overcome this limitation, we developed an alternative strategy employing stereotactic injection of conditional lentiviral vectors (Cre-Flex LVs) encoding Fluc and eGFP in the subventricular zone (SVZ) of Nestin-Cre transgenic mice, thereby specifically labeling the eNSCs. Upon induction of stroke, increased eNSC proliferation resulted in a significant increase in BLI signal between 2 days and 2 weeks after stroke, decreasing after 3 months. Additionally, the BLI signal relocalized from the SVZ towards the infarct region during the 2 weeks following stroke. Histological analysis at 90 days post stroke showed that in the peri-infarct area, 36% of labeled eNSC progeny differentiated into astrocytes, while 21% differentiated into mature neurons. In conclusion, we developed and validated a novel imaging technique that unequivocally demonstrates that nestin+ eNSCs originating from the SVZ respond to stroke injury by increased proliferation, migration towards the infarct region and differentiation into both astrocytes and neurons. In addition, this new approach allows non-invasive and specific monitoring of eNSCs over time, opening perspectives for preclinical evaluation of candidate stroke therapeutics.

【 授权许可】

Unknown