In vitro MTT Assay to Evaluate Mitochondrial Dysfunction in Rat Brain Synaptosomes

Neurodegenerative diseases are a common upshot of synaptosomal mitochondrial malady. The brain synaptosomes are rich in synaptosomal proteins that play an essential part in neuronal activity. Mitochondrial respiratory chain complex dysfunction is a common factor in the generation of free radicals. Neurotoxins such as rotenone led to neuronal cell death as a virtue of a vicious cycle of causing mitochondrial dysfunction, which releases free radicals, and free radicals further lead to more intense mitochondrial damage. The MTT assay is widely used to quantify the viable cells which reduces the MTT into its violet-coloured formazan. The current article focuses on standardizing the acceptable concentration of rotenone, which can be used to screen various test drugs for neuroprotective activity. The four different concentrations i.e., 25, 50, 100 and 200µl of rotenone (1mM) were selected for the study. It was found that 50 and 100µl of rotenone (1mM) was more effective in causing significant and up to 50 percent of mitochondrial disruption. Thus, the invitro MTT assay along with the rotenone induced mitochondrial dysfunction in rat brain synaptosome can be successfully used to screen potential neuroprotective agents.