Abstract
Mitochondria are essential organelles involved in the maintenance of cell growth and function, and have been investigated as therapeutic targets in various diseases. Recent studies have demonstrated that direct mitochondrial transfer can restore cellular functions of cells with inherited or acquired mitochondrial dysfunction. However, previous mitochondrial transfer methods are inefficient and time-consuming. Here, we developed a simple and easy mitochondrial transfer protocol using centrifugation, which can be applied to any cell type. By our simple centrifugation method, we found that the isolated mitochondria could be successfully transferred into target cells, including mitochondrial DNA-deleted Rho0 cells and dexamethasone-treated atrophic muscle cells. We found that mitochondrial transfer normalised ATP production, mitochondrial membrane potential, mitochondrial reactive oxygen species level, and the oxygen consumption rate of the target cells. Furthermore, delivery of intact mitochondria blocked the AMPK/FoxO3/Atrogene pathway underlying muscle atrophy in atrophic muscle cells. Taken together, this simple and rapid mitochondrial transfer method can be used to treat mitochondrial dysfunction-related diseases.
Highlights
Mitochondria are powerful and dynamic organelles responsible for essential cell functions, including energy metabolism, generation of free radicals, maintenance of calcium homeostasis, cell survival and death
A recent study indicates that increased production of mitochondrial reactive oxygen species is a major contributor to mitochondrial damage and dysfunction associated with prolonged skeletal muscle inactivity[6]
We evaluated the effects of mitochondrial transfer on cells with induced mitochondrial dysfunction by treatment with oligomycin[24] and ethidium bromide (EtBr) (Rho[0] cells)[25]
Summary
Mitochondria are powerful and dynamic organelles responsible for essential cell functions, including energy metabolism, generation of free radicals, maintenance of calcium homeostasis, cell survival and death. Increased mitochondrial fragmentation caused by mROS production results in cellular energy stress (e.g., a low ATP level) and activation of the AMPK-FoxO3 signalling pathway, which induces expression of atrophy-related genes, protein breakdown and muscle atrophy[5,6,12]. These results indicate that modulation of mROS production plays a major role in the prevention of muscle atrophy. We compared changes in mitochondrial metabolic function and the signalling pathway from dexamethasone (Dexa)-induced atrophic L6 muscle cells receiving intact mitochondria and damaged mitochondria
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