Neurotoxicity is the capacity of certain compounds to affect the normal activity of the neurons. The mechanism involving the neuron citotoxicity can be several: oxidative stress, DNA damage, cytoskeleton dysfunction or apoptosis. We analyze the following parameters in the Neurotoxicity Assay:
- Cell Viability
- Apoptosis: Caspase 3/7 Activation
- Membrane Damage
- Neurite Outgrowth
- Mitochondrial Damage
In this assay we measure the parameters which frequently appear in the neurotoxicity induced by drugs. To measure the mitochondrial damage we use the TMRM probe and we use inmunocytochemistry for the other three parameters. The Tuj protein is used to mark the neurites and measure the neurite outgrowth. There are compounds that affect the cytoskeleton and disestablish the microtubules affecting the cellular architecture and functionality. We measure the membrane damage by LDH release quantification in the supernatant. Finally the measurement of active caspase 3 shows the capacity of the compound to activate the apoptotic pathway and produce the neuronal death. We use negative controls to compare results of each parameter and when the difference respect to controls of one of the parameters is statistically significant, the drug is neurotoxic.
Cell line used: Primary Neurons
Readout: Neurite Outgrowth, LDH Release, Caspase Activation, Cell Viability