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Innoprot

Ensayo de Neurotoxicidad - High Content

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Neurotoxicity is the capacity of certain compounds to affect the normal activity of the neurons.  The mechanism involved in the neuron citotoxicity can be several: oxidative stress, DNA damage, cytoskeleton dysfunction or apoptosis


Endpoints


Compound Testing


In this assay we measure the parameters which frequently appear in the neurotoxicity induced by drugs. The oxidative stress is measured using the CM probe and the other three parameters are measured by inmunocytochemistry. 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. An antibody against phosporilated H2AX protein is used to measure the DNA damage. The H2AX is a member of the histone H2A family which phosphorilate when the double strand of DNA is broken. The presence of phosphorilated H2AX is correlated with DNA damage and subsequently with genotoxic agents presence. Finally the measurement of active caspase 3 shows the capacity of the compound to activate the apoptotic pathway and produce the neuronal death. These four parameters are measured and compared with negative controls and when the difference respect to controls of one of the parameters is statistically significant, the drug is considered neurotoxic.


 

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