Miro1-mediated mitochondrial positioning shapes intracellular energy gradients required for cell migration

Schuler, Max Hinderk and Lewandowska, Agnieszka and Di Caprio, Giuseppe and Skillern, Wesley and Upadhyayula, Srigokul and Kirchhausen, Tom and Shaw, Janet M. and Cunniff, Brian (2017) Miro1-mediated mitochondrial positioning shapes intracellular energy gradients required for cell migration. Molecular Biology of the Cell, 28 (16). pp. 2159-2169. ISSN 1059-1524 (https://doi.org/10.1091/mbc.E16-10-0741)

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Abstract

It has long been postulated, although never directly demonstrated, that mitochondria are strategically positioned in the cytoplasm to meet local requirements for energy production. Here we show that positioning of mitochondria in mouse embryonic fibroblasts (MEFs) determines the shape of intracellular energy gradients in living cells. Specifically, the ratio of ATP to ADP was highest at perinuclear areas of dense mitochondria and gradually decreased as more-peripheral sites were approached. Furthermore, the majority of mitochondria were positioned at the ventral surface of the cell, correlating with high ATP:ADP ratios close to the ventral membrane, which rapidly decreased toward the dorsal surface. We used cells deficient for the mitochondrial Rho-GTPase 1 (Miro1), an essential mediator of microtubule-based mitochondrial motility, to study how changes in mitochondrial positioning affect cytoplasmic energy distribution and cell migration, an energy-expensive process. The mitochondrial network in Miro1-/- MEFs was restricted to the perinuclear area, with few mitochondria present at the cell periphery. This change in mitochondrial distribution dramatically reduced the ratio of ATP to ADP at the cell cortex and disrupted events essential for cell movement, including actin dynamics, lamellipodia protrusion, and membrane ruffling. Cell adhesion status was also affected by changes in mitochondrial positioning; focal adhesion assembly and stability was decreased in Miro1-/- MEFs compared with Miro1+/+ MEFs. Consequently Miro1-/- MEFs migrated slower than control cells during both collective and single-cell migration. These data establish that Miro1-mediated mitochondrial positioning at the leading edge provides localized energy production that promotes cell migration by supporting membrane protrusion and focal adhesion stability.

  • Item type:Article
    ID code:85670
    Dates:
    Date
    Event
    1 August 2017
    Published
    6 June 2017
    Accepted
    26 October 2016
    Submitted
    Notes:Funding Information: We thank G. Yellen for valuable insight into the use and interpretation of PercevalHR. We thank G. Yellen, A. McKenzie, M. Beckerle, M. Smith, and L. Hoffman for discussion and reporter constructs. We thank M. Redd, S. Hammond, and L. VanderMeer for technical assistance. Spinning-disk confocal fluorescence microscopy was performed in the Kirchhausen laboratory, with technical support from J. Houser. Lattice light sheet microscopy was performed in the Kirchhausen laboratory by S.U., W.S., G.D., and B.C. Seahorse extracellular flux analysis was performed at the University of Utah Metabolic Phenotyping Core with technical assistance from A. Laxman. Wide-field and scanning confocal fluorescence microscopy was performed at the University of Utah Fluorescence Microscopy Core (NCRR Shared Equipment Grant 1S10RR024761-01). This work was funded by National Institutes of Health Grants GM84970 to J.M.S. and GM075252 to T.K. Construction of the lattice light-sheet microscope was supported by grants from Biogen and Ionis Pharmaceuticals to T.K. Publisher Copyright: © 2017 Schuler et al. Miro1-mediated mitochondrial positioning shapes intracellular energy gradients required for cell migration, Max-Hinderk Schuler, Agnieszka Lewandowska, Giuseppe Di Caprio, Wesley Skillern, Srigokul Upadhyayula, Tom Kirchhausen, Janet M. Shaw, and Brian Cunniff Molecular Biology of the Cell 2017 28:16, 2159-2169, https://doi.org/10.1091/mbc.e16-10-0741
    Subjects:Science > Microbiology
    Department:Faculty of Engineering > Biomedical Engineering
    Depositing user: Pure Administrator
    Date deposited:01 Jun 2023 14:51
    Last modified:21 Apr 2024 02:59
    Related URLs:
    URI:https://strathprints.strath.ac.uk/id/eprint/85670
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