Analysis of mitochondrial network by imaging: proof of technique in schizophrenia.

Mitochondria, similar to living cells and organelles, have negative membrane potential and can therefore accumulate permeable lipophilic cations. Those cations which exhibit fluorescence activity after accumulation into energized systems are widely used to decipher changes in membrane potential by imaging techniques. Here we describe the use of the lipophilic cation 5,5',6,6'tetrachloro-1,1',3,3'-tetraethylbenzimidazol-carbocyanine iodide (JC-1), which alters reversibly its color from green (J-monomer, at its low concentration in the cytosol) to red (J-aggregates, at its high concentration in active mitochondria) with increasing mitochondrial membrane potential (Δψm). We show that in addition to changes in Δψm, this specific dye can be used to follow alterations in mitochondrial distribution and mitochondrial network connectivity. We suggest that JC-1 is a preferable probe to compare between treatment groups, as the ratio of green to red fluorescence intensities is used for analysis. This ratio depends only on the mitochondrial membrane potential and not on other mitochondrial dependent or independent factors. We demonstrate various applications of JC-1 staining to study mitochondrial abnormalities in different cell types derived from schizophrenia patients and healthy subjects.