Study Multiple Parkinson's Disease Mechanisms of Action in Parallel
The multiple pathways implicated in Parkinson’s disease is a complex obstacle to untangle in developing an effective therapeutic. Charles River has developed high content screening phenotypic in vitro assays to provide quantitative data for TOM20 loss and apha-synuclein aggregation.
- Alpha-Synuclein Expression and Aggregation Quantification
Developed in collaboration with The Michael J. Fox Foundation’s Parkinson’s Disease Research Tools Consortium, this assay measures alpha-synuclein aggregation in Parkinson's Disease cell model using an aggregate selective anti alpha-synuclein antibody (MJFR14).
Detect & Measure Primary Biomarkers of Parkinson’s Disease
- Neuronal Mitophagy Assays
Mutations in genes encoding proteins involved in targeted autosomal destruction of mitochondria (e.g., Parkin and PINK1) have been described in PD. This sparked heavy interest in therapeutic approaches that facilitate neuronal mitophagy. High-content imaging methods are used to measure different readouts of mitophagy:
- Tom20 Loss Quantification
Targeting the mitophagic regulators has gained momentum as a therapy based on their link to the gene mutations Parkin and PINK1. This assay measures the loss of the structural mitochondrial marker TOM20 (TOMM20).
- Fluorescent reporters that measure pH changes during mitophagy
- Mitochondrial membrane potential changes as a measure of viability
- Parkin translocation to mitochondria
- Tom20 Loss Quantification
- MPP+ Neuronal Cell Death Assay
N-Methyl-4-phenylpyridinium Iodide (MPP+) is a neuron selective toxin that induces Parkinsonism in animal models. An active (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) MPTP metabolite, MPP+ has been shown to interfere with oxidative phosphorylation and induce mitochondrial dysfunction. This dysfunction results in depleted mitochondrial ATP production, and ultimately, death of neurons. Using high content imaging, we can monitor cell death by measuring levels of propidium iodide that selectively accumulates in the dead neurons.
High Content-Based Assay to Measure TOM20 Loss
Improve Translation Success with Parkinson’s Disease Relevant Cell Models
A variety of neural cell lines can be utilized to perform phenotypic screens. These range from human immortalized neural cell lines derived from the ventral mesencephalon (midbrain) region of the brain to primary rodent neurons to iPSC-derived neuronal models.
These models are readily used for Parkinson’s disease based upon growth characteristics, differentiation potential and permissiveness to viral transduction. In vitro differentiations are performed in-house by depriving particular growth factors and supplementing with others. Differentiations for each phenotypic screen are routinely characterized for mature neuron phenotypes such as relevant gene and protein expression as well as functionality by electrophysiology.
Translation to PD In Vivo Models
Charles River offers comprehensive and readily translatable modeling platforms of chemically, biologically and genome engineered animal models to study mechanism of action and efficacy of novel PD therapies. This portfolio includes established alpha-synuclein, unilateral 6_OHDA lesion, and MPTP lesion models for Parkinson’ disease in vivo studies to better inform preclinical and clinical outcomes.