PARKINSON

In vitro

  • Primary mesencephalic neuron cultures with immature astrocytes able to specifically kill dopaminergic neurons in different medium
  • Primary mesencephalic neuron cultures with / without astrocytes
    Treatment with MPP+, rotenone
  • Primary mesencephalic neuron cultures with / without microglial cells
    Compounds with potential anti-inflammatory activities could be tested (several treatments possible depending on the pathway of interest)
  • Primary cell cultures of cortical, striatal and dopaminergic neurons infected with virus vectors containing aggregated a-synuclein

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Cell survival, cell proliferation, morphological changes, protein measurement, etc.

cell survival, cytokine measurement, cell proliferation, morphological changes, protein density

cell survival, aggregation

Rodents

  • MPTP mouse and 6-OHDA mouse/rat
  • Transgenic models of alpha-synuclein mice, KO Parkin, LRRK2 G2019S mice
  • Nigrostriatal synucleinopathy: viral vector-mediated models (CAV2- mice, AAV-rats) coding for alpha-synuclein (WT or A53T)
  • A “double hit” rat model: co-expression of alpha-synuclein and C-Terminal domain of LRRK2-G2019S (increase of alpha-synuclein toxicity).
Read-out
Behavioral changes, stereological cell count in the substantia nigra (TH-positive neurons), number of alpha-synuclein aggregates, partial loss of dopaminergic fibers/DAT binding sites using microPET radiotracers, etc.

Non-human primates

  • MPTP-lesion model in squirrel monkey, macaque or baboons
  • Nigrostriatal synucleinopathy: viral vector-mediated models (CAV2, AAV) coding for alpha-synuclein (WT or A53T)
Read-out
Behavioral changes, stereological cell count in the substantia nigra (TH-positive neurons), number of alpha-synuclein aggregates, partial loss of dopaminergic fibers/DAT binding sites using microPET radiotracers, etc.

Humans

  • Clinical investigation Center (Phase I and II), Biological resource center, stratified cohorts of patients
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PET, SPECT and MRI imaging, Markers of dopaminergic system and neuroinflammation, platforms of movement analysis, Neurophysiology, abnormal movement recordings, transcranial magnetic stimulation, etc.
A comprehensive overview of our offer in the field of PARKINSON disease:
  • Basic Research

    Cellular models

    In-vitro cell culture (primary cell cultures) of (mouse/rat) dopaminergic (mesencephalic) neurons (with or without astrocytes)

     - Evaluation (through quantification of neuronal survival) of the neuroprotective properties of new molecules in a spontaneous cell death model (neuronal degeneration in the presence of proliferating astrocytes).

    - Evaluation (through quantification of neuronal survival) of the neuroprotective properties of new molecules in a cell death model induced by different neurotoxins (MPTP, rotenone, iron mediated oxydative stress,…)

    In-vitro co-culture (primary cell cultures) of (mouse/rat) dopaminergic (mesencephalic) neurons and microglial cells

    - Evaluation (through quantification of neuron survival and inflammatory activity of microglia) of the anti-inflammatory and/or immunoregulatory properties (cytokines detection and measures of the nitrite levels) of new molecules

    In-vitro model of human dopaminergic cell line (SH-SY5Y cells)

    - Evaluation (through quantification of neuron survival) of the neuroprotective properties of new molecules in a spontaneous cell death model

    In-vitro model of human dopaminergic iPS cells

    - In vitro differentiation of cell lines issued from patients bearing specific mutations implicated in familial forms of PD ( parkin, pink 1) and from healthy controls

    - o Reprogrammation of fibroblasts issued from patients bearing specific mutations implicated in familial forms of PD ( parkin, pink 1, LRRK2) and from healthy controls.

    References :

    Kinugawa K, Monnet Y, Béchade C, Alvarez-Fischer D, Hirsch EC, Bessis A, Hunot S. (2013) DAP12 and CD11b contribute to the microglial-induced death of dopaminergic neurons in vitro but not in vivo in the MPTP mouse model of Parkinson's disease. J Neuro-inflammation, 10(1):82

    Animal models

    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 out1

    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)s

  • Preclinical Research
  • Clinical Research