Animal models and associated analysis tools
Zebrafish Model
Rodent Models (mouse, rat)
In vivo rodent Models of dopaminergic cell degeneration induced by neurotoxins
- Quantification of the neuroprotective effects and/or plastic effects of therapeutic agents in the MPTP mouse model
- These models consists of i.p. injections of MPTP (using either acute, sub-acute or chronic intoxication regimen) which are known to induce an oxidative stress, a mitochondrial deficit and a neuroinflammation
- Evaluation of the neuronal cell death, neurochemical deficits, motoric alterations, and neuroinflammation.
- Possibility of combining the MPTP intoxication with deleterious environmental factors (peripheric inflammation, stress situations, etc…)
Monitoring the anti-parkinsonian effects of potential therapeutic agents in the 6-OHDA (mouse/rat) lesion model :
- Unilateral stereotactic injections of 6-hydroxydopamine (6-OHDA) in the mouse/rat nigrostriatal pathway leading to motor disfunctions.
- 2 models are available :
* intra-nigral injections, leading to a complete lesion model
* intra-striatal injections (dorsal striatum), leading to a partial and more progressive lesion model
- Behavioural analyses
* Apomorphine rotation test, evaluating the severity of the lesion through the number of rotations performed by the lesioned animals following apomorphine i.p. administration
* video-based analyses (CATWALK, Noldus Equipments)
* semi-quantitative analyses of motor symptoms using clinical rating scales and specialized softwares (THE OBSERVER, Noldus Equipments)
Rodent models of nigrostriatal alpha-synucleinopathy induced by viral vector-mediated gene transfer.
- Quantification of the neuroprotective effects and/or plastic effects of therapeutic agents against alpha-synucleinopathy induced after intrastriatal injection of CAV2 viral vectors encoding various forms of alpha-synuclein (wild-type, mutated, truncated) Intrastriatal stereotactic injections of CAV2 vectors (hSyn, hSynA53T/A30P, hSyn110) leading to a progressive degenerative synucleinopathy of the nigrostriatal pathway (1-4 months)
- Evaluation of the neuronal cell death, the protein aggregation level and the neuroinflammatory reaction induced,
Monitoring transcriptomics, biochemical, histological and behavioural alterations and various imaging marker modifications after alpha-synuclein overexpression (hSyn, hSynA53T/A30P) (under development and characterization)
- Intra-nigral stereotactic injections of lentiviral vectors expressing various constructs (hSyn, hSynA53T/A30P, hSyn110) leading to a progressive (slowly degenerative) synucleinopathy of the nigrostriatal pathway (1-4 months)
- Evaluation of the neuronal cell death, the protein aggregation level and the neuroinflammatory reaction produced in these models and their modifications upon experimental treatments
In vivo rodent models of parkin and pink1 knock out
In vivo prodromal model associated with nigral overexpression of various truncated forms of the mutated LRRK2 protein
- Transcriptomic alterations (biomarkers) associated with a specific mutation (G2019S) of the LRRK2 protein (under development and characterization)
Non-human Primate models
MPTP-lesion model in Saïmiri sciureus (squirrel monkey)
- Evaluation and quantification of the neuroprotective effects and/or plastic effects and/or anti-parkinsonian effects of therapeutic agents in this primate model of PD
- Model of chronic MPTP intoxication until a stable parkinsonian state is reached.
- Behavioral analyses (clinical rating scale for motor symptoms, video-tracking analysis)
- Longitudinal analyses on biological fluids (plasmatic serum, CSF)
- Neuropathological analyses (neuronal cell death markers, neuroinflammation, neurochemical alterations,…).
In vivo model of dopamine cell degeneration through chronic systemic MPTP injections in Macaca fascicularis (macaque cynomolgus) or Papio anubis (baboons).
2 models are available :
- Acute intoxication paradigm (around 2 month duration)
* Stable parkinsonian state
* Severe model, up to 90% striatal deafferentation, mimicking a end-stage state of the pathology
* Can be used to induce L-DOPA-induced dyskinesias (model also at disposal)
* particularly adapted to preclinical trials of neuroreplenishment strategies or neurotrophic-like molecules
- Chronic intoxication paradigm (6-12 month duration)
* stable parkinsonian state
* Model of progressive striatal deafferentiation (Putamen>Caudate nucleus)
* Particularly adapted to preclinical trials of neuroprotective and/or neurotrophic strategies
Evaluation of therapeutic efficacy and post mortem characterisation
Rodents
in vivo imaging
PET Imaging
- PET parametric imaging of the dopaminergic system in the 6-OHDA rat lesion model
* Dopaminergic presynaptic markers ( DA transporter) : 18F-LBT, 11C-PE2I
* Dopaminergic post-synaptic markers (D2-receptors) : 18F-fallypride, 11C-raclopride
- PET parametric imaging of the regional metabolic rate of glucose : 18F-FDG
- PET parametric imaging of the neuro-inflammation : 18F-DPA714
- Compartmental modeling and parametric imaging, automatized input function, PET/MRI co-registration (neuroFocus 220/ MRI 11.7T)tomatisée, TEP/IRM co-registration (neuroFocus220/IRM 11.7T)
Very high field MR imaging / MR spectroscopy of rodents (11.7 T Bruker)
- T2-weighted imaging
- DTI
- Proton NMR spectoscopy, quantification through LC Model
- Glutamate CEST-imaging
- Diffusion NMR Spectroscopy of various NMR detectable metabolites (NAA, Glutamate, Glutamine, etc…).
Rodents: histopathological analyses
- Immunohistochemical detection of various molecular targets
* on free floating sections
* on sections immunostained using a Ventana automate (high standardization)
* paraffine sections on Ventana automate
- Confocal immunofluorescence
- Unbiased Stereological counts
- 3D-reconstruction of whole brain serial histological sections
- In vivo/post-mortem co-registration of reconstructed histological volumes with in vivo PET and/or MR imaging volumes
- Reglementary anatomo-pathological studies (rodent/primate) by a trained anatomo-pathologist (European board bearer)
PRIMATES
In vivo imaging
Very high field MR imaging / MR spectroscopy of non-human primates (7 T) – can be performed under BSL1/BSL2/BSL3 conditions
- T2-weighted imaging
- Proton NMR spectoscopy, quantification through LC Model
- Glutamate CEST-imaging
- Diffusion NMR Spectroscopy of various NMR detectable metabolites (NAA, Glutamate, Glutamine, etc…).
PET Imaging
- PET parametric imaging of the dopaminergic system in the 6-OHDA rat lesion model
* Dopaminergic presynaptic markers ( DA transporter) : 18F-LBT, 11C-PE2I
* Dopaminergic post-synaptic markers (D2-receptors) : 18F-fallypride, 11C-raclopride
- PET parametric imaging of the regional metabolic rate of glucose : 18F-FDG
- PET parametric imaging of the neuro-inflammation : 18F-DPA714
- Compartmental modeling and parametric imaging, PET/MRI co-registration (neuroFocus 220/ MRI 7T)
Other material and methods
Intracerebral administration of substances/vectors under stereotactic control
- assisted through per-operative MRI and/or
- neuronavigation (MEDTRONICS neuro-navigator). n
Behavioural analyses
- motor functions
* Video movement analysys (ETHOVISION, Noldus Equipments)
* Semi-quantitative (clinical scoring) analysis of various symptoms (THE OBSERVER, Noldus Equipments) 11.7T)
- cognitive functions
* tests on tactile screens performed in individual cages and/or in gang (social groups)
* tests include the full CANTAB Battery for NHP (DNMS, DMS, IEDS, CSST, Memory 30, Visual Discrimination Task, etc…
* As well as some built in-house batteries of cognitive tests that can be adapted for specific purposes
Longitudinal studies on biological fluids (blood plasma, CSF, …)
Histopathological analyses
Immunohistochemical detection of various molecular targets
- on free floating sections
- on sections immunostained using a Ventana automate (high standardization)
- on paraffine sections and Ventana automate
Confocal immunofluorescence
Unbiased Stereological couts
3D-reconstruction of whole brain serial histological sections
In vivo/post-mortem co-registration of reconstructed histological volumes with in vivo PET and/or MR imaging volumes
Reglementary anatomo-pathological studies (rodent/primate) by a trained anatomopathologist (European board bearer)