Mitochondria are dynamic organelles that undergo size and morphological changes due to fusion/fission events, transport, mechanical stress, and signaling cascades, all of which impact their function. The cytoskeleton, comprising microtubules, actin filaments (F-actin), and intermediate filaments (IFs), forms a network that exerts mechanical forces on cellular components like mitochondria, influencing their morphology and function. Molecular motors (kinesin, dynein, and myosin) bind to mitochondria, affecting their shape and transport along cytoskeletal filaments.

While interactions between mitochondria and the cytoskeleton alter mitochondrial function, the underlying mechanisms are not fully understood. In this study, we examined these interactions in Xenopus laevis melanophore cells expressing fluorescent microtubules. We selectively disrupted cytoskeletal networks and analyzed their effects on mitochondrial transport, organization, morphology, and shape fluctuations using super-resolution Airy-scan and confocal microscopy. Individual mitochondria were tracked with nanometer precision.

Our results show that microtubules play a dominant role in mitochondrial organization and transport, while F-actin and vimentin IFs influence organelle morphology. Mechanical interactions between cytoskeletal filaments and mitochondria shape their behavior, and motor proteins are critical for their transport. These findings highlight key aspects of mitochondria-cytoskeleton interactions and their potential role in mitochondrial function and cellular homeostasis.