The effect of respiration buffer composition on mitochondrial metabolism and function

摘要: Functional studies on isolated mitochondria critically rely the right choice of respiration buffer. Differences in buffer composition can lead to dramatically different rates leading difficulties comparing prior studies. The ideal facilities high ADP-stimulated respiratory and minimizes substrate transport effects so that ability distinguish between various treatments conditions is maximal. In this study, we analyzed a variety buffers combinations determine optimal support mitochondrial function through uncoupled using FCCP. consisted standard KCl based (B1) three modified with chloride replaced by K-lactobionate, sucrose, antioxidant taurine (B2) or K-gluconate (B3). fourth (B4) was identical B2 except K-lactobionate K-gluconate. metabolites utilize pathways metabolism. To test function, used cardiac guinea pig measured oxygen consumption for states an Oroboros Oxygraph-2k. These were leak state (energized absence adenylates), presence saturating ADP concentrations), FCCP). On average across all combinations, B2, B3, B4 had increase 16%, 26%, 35% state, ADP-simulated respectively, relative B1. common feature distinguishing these from B1 notable lack concentrations. Based rate metrics obtained conclude adenine nucleotide translocase, dicarboxylate carrier, alpha-ketoglutarate exchanger are partially inhibited chloride. Therefore, when goal maximize respiration, containing superior choices compared KCl-based buffers.