Parkinson’s Disease In Vitro Assays: Study Multiple MOAs in Parallel
Parkinson’s disease requires potential therapeutics to be studied against several mechanisms of action (MOA). Charles River has developed Parkinson’s disease in vitro assays to identify and validate compounds through phenotypic screens and profiling. Using high content imaging, you can quantify leading Parkinson’s biomarkers (proteins, alpha-synuclein aggregation and mitophagy,TOM20) in relevant cell populations for predictive translation to PD studies.
- Protein Biomarkers
Proteins thought to influence cell homeostasis or neural death have been implicated in Parkinson's disease progression. Measuring the changes in these protein levels via high content imaging can be used to identify novel PD therapeutics.- Alpha-Synuclein Expression and Aggregation Quantification - Developed in collaboration with The Michael J. Fox Foundation’s Parkinson’s Disease Research Tools Consortium, this Parkinson’s disease in vitro assay measures alpha-synuclein aggregation in a PD cell model using an aggregate selective anti alpha-synuclein antibody (MJFR14).

Detect and Measure Primary Biomarkers of Parkinson’s Disease
- Neuronal Mitophagy Assays
Mutations in gene 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 Parkinson’s disease in vitro 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
- 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, resulting in depleted mitochondrial ATP production, and ultimately, the 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 Mitophagy TOM20 Loss
Frequently Asked Questions about Parkinson’s Disease in Vitro Assays
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Are there disease-relevant cells lines for use in Parkinson’s Disease in vitro assays?
One example of PD relevant cells is differentiated ReNcells, which represent a replenishable source of human dopaminergic neurons. These have been developed for use in Parkinson’s biomarker assays (Alpha-Synuclein Expression & Aggregation Quantification) and Mitophagy Assays (TOM20 Loss). High Content Imaging of these differentiate ReNcells provide predictive translational data.
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What are the advantages of using high content imaging for Parkinson’s disease research?
HCI format provides phenotypic data for specific targets and can bring to light off-target effects in the same assay. These phenotypic Parkinson’s disease in vitro assays can be optimized and scaled for high-throughput screening of potential Parkinson disease therapeutics, speeding the drug discovery process.