Bioluminescence platform

Scientific Advisor: Jasna Kriz.

Background information
Over the past years the bioluminescence platform has generated and validated several transgenic mice expressing dual- bioluminescence/ fluorescence reporters, the firefly luciferase (Fluc) and GFP, whose transcription will be dependent upon the selected gene promoter. The rationale and the advantage for the use of the dual reporter system emerged from the fact that fluorescence signals can be used to achieve microscopic resolution (with the microscopes equipped with two-photon laser scanning capabilities) while bioluminescence, owing to favourable emission spectra of luciferase (above 620 nm) is optimized for whole-body non-invasive imaging (Lalancette et al., 2009, Gravel et al., 2011). Here it is important to mention that quantification of the in vivo light production will reveal the spatial and temporal expression of the selected gene promoters in the living mice. The developed in vivo imaging model systems target processes such as inflammation, neuronal damage, axonal regeneration attempt and neurogenesis. Importantly, the high homology between the mouse and human genomes permits to study functional implication of certain genes and its promoter activities in distinct pathological conditions. Our previous studies clearly suggest that biophotonic signals imaged from the live animals can be used as valid biomarkers to screen for novel biocompatible molecules (Maysinger et al., 2007, Hutter et al., 2010, Lalancette et al, 2010) and/or to visualize distinct pathological events in acute injuries such as brain icshemic (Lalancett et al., 2009, 2011,2012), and in chronic neurological disorders including, ALS and Frontotemporal dementia, ALS (Keller et al.,2009, 2011, Swarup et al., 2011). The basic principle for in vivo bioluminescence imaging are schematically presented in Figure 1. Our in vivo imaging system (IVIS 200) allows 2 D and 3D in vivo imaging Figure 2.

Table 1: Available & validated reporter mouse models:

Transgenes Imaging applications
Tlr2 promoter/Fluc-GFP Microgliosis
(inflammation)
GFAP promoter/Fluc Astrogliosis (Caliper/PE)
(inflammation)
Gap-43 promoter/Fluc-GFP Neuronal response to injuries and regenerating axons
nestin (DCX) promoter/Fluc-GFP Neurogenesis

Applications: In vivo pathology, In vivo pharmacology

Available disease models - expertise present in the bioluminescence platform
Injury models:

  • stroke models (transient and permanent middle cerebral artery occlusion,
  • peripheral nerve injuries: sciatic nerve cut and crush injuries

Genetic disease models crossed with reporter mice ( double transgenic reporter mice)

  • SOD1G93A and SOD1G37R transgenic mice ( mosue models of ALS)
  • TDP-43WT and TDP-43mutants (mouse models of Frontotemporal dementia/ALS)
  • APP/PS1 transgenic model of Alzhemier’s disease

Services provided within the platform;

  • bioluminescnce imaging training and experimental design
  • in vivo disease model-system validation
  • in vivo pathology analysis
  • in vivo pharmacology -targeted drug screening possibilities using available reporter mouse models

Personnel:
Yuan Cheng Weng, MS, DVM (Senior Research Assistant)

Relevant publications:

  1. Cordeau P, Kriz J.(2012) Real-time imaging after cerebral ischemia: model systems for visualization of inflammation and neuronal repair. Methods Enzymol. 2012;506:117-33
  2. Swarup V, Audet JN, Phaneuf D, Kriz J, Julien JP (2012). Abnormal Regenerative Responses and Impaired Axonal Outgrowth after Nerve Crush in TDP-43 Transgenic Mouse Models of Amyotrophic Lateral Sclerosis. J Neurosci. 12;32:18186-95
  3. Swarup V, Phaneuf D, Bareil C, Roberston J, Kriz J*, Julien JP*(2011). Pathological hallmarks of ALS/FTLD in transgenic mice produced with genomic fragments encoding wild-type or mutant forms of human TDP-43. Brain Pt 9:2610 * co-corresponding authors
  4. Lalancette–Hébert M, Julien C, Cordeau P, Bohacek I , Weng YC, Calon F, Kriz J (2011). Accumulation of Dietary DHA in the Brain Attenuates Acute Immune Response and Development of Post-ischemic Neuronal Damage. Stroke 10:2903
  5. Gravel M, Weng YC, Kriz J (2011). Mouse model for live imaging of neuronal response to injury and repair. Mol Imag 10(6):434-45
  6. Keller F, Gravel M, Kriz J (2011). Treatment with minocycline after disease onset alters astrocyte reactivity and increases microgliosis in SOD1 mutant mice Exp Neurol 228:69
  7. Lalancette-Hébert M, Moquin A, Choi A, Kriz J*, Maysinger D* (2010) Lipopolysaccharide-QD micelles induce marked induction of TLR2 and lipid droplet accumulation in olfactory bulb microglia. Molecular Pharmaceutics 2010 May 21. [Epub ahead of print]
  8. .First author is my graduate student, *co-corresponding authors.
  9. Hutter E, Boridy S, Labreque S, Lalancette-Hébert M, Kriz J, Winnik F, Maysinger D. (2010) Microglial uptake and response to gold nanoparticles: The effects of nanomorphology and surface. ACN Nano, 2010 May 25;4(5):2595-606
  10. Lalancette-Hébert, M., Phaneuf, D., Soucy, G., Weng, Y.C. and Kriz, J. (2009) Live imaging of Toll-like receptor 2 response in cerebral ischaemia reveals a role of olfactory bulb microglia as modulators of inflammation. Brain;132:940-54.
  11. Cordeau Jr. P*, Lalancette-Hébert M*, Weng YC, Kriz J (2008). Live imaging of neuroinflammation reveals gender and estrogen effects on astrocyte response to ischemic injury. Stroke, 39, 935-42