MPP+ Neuronal Cell Death Assay

Parkinson's disease is one of the most common neurodegenerative diseases, and its main pathological features are the impairment of motor function and neurological function caused by dopamine degeneration. The onset of Parkinson's disease or its pathological symptoms can be induced in vitro or in vivo by a variety of methods, such as the use of Parkinson's disease inducers, modeling using related proteins, or direct use of transgenic cell lines with pathogenic mutations. Among them, 1-methyl-4-phenyl-1,2,3,6-tetrahydro-pyridine (MPTP) and its active metabolite 1-methyl-4-phenylpyridinium (MPP+) can ablate dopaminergic and mildly perturb motor ability in various animal species including non-human primates, and also show the highest survival rate of cells after modeling, which is currently recognized as an effective Parkinson's Disease Inducer. At Creative Biolabs, we provide stable and efficient MPP+ induction modeling, cell death assays and other related tests to pave the way for your research.

Background and Mechanism of MPP+

MPTP is a molecule structurally similar to the herbicide paraquat, and MPP+ is its active metabolite. MPTP and MPP+ are the most commonly used toxins for in vitro models of Parkinson's disease, which produce clinical, biochemical and pathological changes similar to those of common Parkinson's disease. Prolonged exposure to low MPTP or MPP+ doses promotes increased oxidative stress, α-syn fibrillation, and loss of mitochondrial function.

MPTP selectively damages dopaminergic neurons in the brain, and it is a highly lipophilic compound in nature, giving it the ability to cross the blood-brain barrier. MPTP itself is not a toxic substance, but after entering the brain, it will be metabolized into 1-methyl-4-phenyl-2,3-dihydropyridine (MPDP) by non-monoamine oxidase B and captured by astrocytes. The MPDP is then oxidized to the active and biotoxic MPP+ and released outside the cell. Extracellular MPP+ is actively taken up by high-affinity dopamine transporter (DAT) and stored in dopamine vesicles, where it induces K+ efflux, promotes membrane hyperpolarization, blocks the activity of complex I activity, leads to ATP depletion and reactive oxygen species production, which ultimately causes cell death.

Fig 1. Mechanism of MPP+ toxicity. (Risiglione, et al., 2020)

Our MPP+ Neuronal Cell Death Assays

In recent years, there has been a lot of evidence that MPP+ induced mitochondrial dysfunction and oxidative stress are involved in the pathological mechanism of PD, and MPP- induced non-apoptotic cell death has characteristics similar to ferroptosis.

We provide commonly used experimental models for MPP-mediated cytotoxicity studies, such as green fluorescent protein transgenic zebrafish, rat pheochromocytoma (PC12) cell line, and SH-SY5Y cells. The measurement of intracellular reactive oxygen species can be achieved by the fluorescent probe 2,7-dichlorofluorescein diacetate (DCF-DA). DCF-DA can be oxidized to DCF by intracellular hydrogen peroxide or other low molecular weight peroxides and quantitative measured by fluorescence. Cytotoxicity can be assessed by cell counting kit-8 (CCK-8). In addition, a series of classical immunology and fluorescence imaging methods are also optional approaches.

Fig 2. Fluorescence detection of intracellular reactive oxygen species. (Lu, et al., 2015)

Thanks to our professional R&D and experimental team, Creative Biolabs performs Parkinson's Disease MPP+ Assay with dopaminergic neuronal cell cultures, and we can provide services such as establishing MPP+-induced in vitro models, verifying cell viability and mortality, identifying drug effects or analyzing its cytotoxicity. If you have any need or want to further advance your research, please do not hesitate to contact us.

References

1. Risiglione, P.; et al. High-Resolution Respirometry Reveals MPP+ Mitochondrial Toxicity Mechanism in a Cellular Model of Parkinson's Disease. International Journal of Molecular Sciences. 2020, 21(21): 1-15.
2. Lu, X.L.; et al. Paeonolum protects against MPP+ induced neurotoxicity in zebrafish and PC12 cells. BMC Complementary and Alternative Medicine. 2015, 15: 137.