For a cell to function and survive following a period of physiological stress, it needs to be able to meet the energy demand required to restore homeostasis. In our lab, we seek to identify how mitochondria respond and adapt to nutritional stressors to better understand how mitochondria mediate the risk of developing disease. The current methods for testing PBMC bioenergetics involve measuring respiratory capacity after adding inhibitors and uncouplers.
These methods have helped us understand significant bioenergetic changes in PBMCs in different disease states. Membrane potential is essential for ATP synthesis and regulates processes such as respiratory flux, reactive oxygen species, and autophagy. However, we still have limited knowledge about how mitochondrial respiration and membrane potential together respond to physiological substrate concentrations in PBMCs.
The advantage of this technique is that it allows for an integrated analysis of mitochondrial membrane potential and oxygen consumption in human PBMCs in response to increasing levels of ADP. This method allows for the quantification of the sensitivity of mitochondria to a shift in energy demand.
