Hi,
I am working on Leishmania parasites and following treatment with some drugs, I wanted to measure the health of the mitochondria. For this, I used JC-1 dye. The drugs that I use interfere with sterols. The JC-1 staining data is kind of surprising. I have attached a picture of the data. On left is the control and on right is the drug treated sample. As you can see compared to the controls, there are two distinct populations in the drug treated sample. I am not sure whether this indicates a hyperpolarization of the mitochondria? By the way, these parasites have only one large mitochondrion per cell.
As we know that mitochondrial hyperpolarization would lead to a decrease in ATP synthesis, I measured the cellular ATP as well. Particularly, I measured total ATP, ATP from glycolysis and ATP from mitochondrial oxidation. To my surprise, in all cases, I see a marked increase in drug treated parasites than the controls. I was wondering, if these dot plots truly represent hyperpolarization or something else? If it is true, then how come membrane hyperpolarization leads to an increase in both glycolytic and mitochondrial ATP production? And finally, do such things represent early apoptotic event?
Any comment will be highly appreciated.
Thanks Sumit
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