Primary E18 rat hippocampal neurons were cultured on poly-lysine coated glass coverslips until DIV8. Next, neurons were treated with FCCP, a protonophore and uncoupler of oxidative phosphorylation in mitochondria, for a defined period of time. Labeling of neurons with MitoTracker Red indicates active mitochondria. After fixation, neurons were labelled for MAP2. The effect of compounds on active mitochondria in soma versus neurites can be evaluated in individual neurons using Image Pro Plus software.
Figure: Effect of FCCP on active mitochondria in primary hippocampal rat neurons. FCCP treatment significantly reduced MitoTracker Red labeling in somata and neurites of primary rat hippocampal neurons (see graph). Images indicate (left) control neurons and (right) neurons upon FCCP treatment. MitoTracker labeling is indicated in red, MAP2 labeling in green.
Remark: This assay can also be performed with other mitochondria related toxins.