I have evidence of mitochondrial dysfunction (Seahorse assay) but no evidence for an increase in mitochondrial mass (fluorescence assay). What might be causing the dysfunction, particularly increased ATP production, if not an associated increase in mitochondrial mass?
Hi David. have you measure ATP levels over time or at a single time point? If the latter is true, than perhaps there is less ATP-ase activity and you have more unused ATP left over as a result.
Hi Vedad, thank you for your response. I have just one time point measurement for ATP production. Thanks for the ATP-ase tip, I'll look into that. cheers!
I agree with Vedad - you are inhibiting some major process that is using ATP. Primary ion transporters maintaining ion gradients across the membrane -like Na+/K+-ATPase- are first that come to mind because they use up to 80% of cellular ATP, depending on cell type. Unfortunately, with your question being short on details (is it cells or purified mitochondria, which part of the Seahorse profile is affected and so on), it is hard to be more precise. Hope it helps
Mitochondria are not solely ATP providers they have a lot of other important roles (apotose induction , ROS production, mitophagy etc) perhaps the conditions in your assays induced a shift in roles. Mitochondria are extremely flexible structure changers and supercomplex reassembling may induce higher or lower energetic efficiency (ATP production) without affecting mitochondrial mass. They may also be limited at the electron entry level at complex I, II G3PDH or others electron entry pathways. So even if they high ATP generation capacities they may not receive all the electrons needed.
As our colleagues above already mentioned Additional data would help to pinpoint possible causes of the phenomenon you observe.
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