Mitochondria are morphologically heterogeneous within cells

Mitochondria play key roles in the life and death of cells. We investigated whether mitochondria represent morphologically continuous entities within single intact cells. Physical continuity of mitochondria was determined by three-dimensional reconstruction of fluorescence from mitochondrially targeted DsRed1 or tetra-methyl rhodamine ethyl ester (TMRE). The mitochondria of pancreatic acinar, porcine aortic endothelial (PAE) cells, COS-7 cells and SH-SY5Y cells and neocortical astrocytes all displayed heterogeneous distributions and were of varying sizes. In general, there was a denser aggregation of mitochondria in perinuclear positions than in the cell periphery, where individual isolated mitochondria could clearly be seen. DsRed1 was found to be highly mobile within the matrix of individual mitochondria, with an estimated linear diffusion rate of 1 mum s(-1). High-intensity irradiation of subcellular regions bleached the fluorescence of mitochondrially targeted DsRed1, but did not cause the mitochondria to depolarise or fragment. A lack of rapid fluorescence-recovery-after-photobleaching (FRAP) of DsRed1 indicated lumenal discontinuity between mitochondria. We observed a slow (half-time approx. 20 min) recovery of DsRed1 fluorescence within the irradiated area that was attributed to mitochondrial movement or fusion of unbleached and bleached organelles. Mitochondria were not electrically coupled, since typically only individual mitochondria were observed to depolarise following irradiation of TMRE-loaded cells. Our data indicate that the mitochondria within individual cells are morphologically heterogeneous and unconnected, thus allowing them to have distinct functional properties.