Letter | Published:

Mitochondrial reticulum for cellular energy distribution in muscle

Nature volume 523, pages 617620 (30 July 2015) | Download Citation

Abstract

Intracellular energy distribution has attracted much interest and has been proposed to occur in skeletal muscle via metabolite-facilitated diffusion1,2; however, genetic evidence suggests that facilitated diffusion is not critical for normal function3,4,5,6,7. We hypothesized that mitochondrial structure minimizes metabolite diffusion distances in skeletal muscle. Here we demonstrate a mitochondrial reticulum providing a conductive pathway for energy distribution, in the form of the proton-motive force, throughout the mouse skeletal muscle cell. Within this reticulum, we find proteins associated with mitochondrial proton-motive force production preferentially in the cell periphery and proteins that use the proton-motive force for ATP production in the cell interior near contractile and transport ATPases. Furthermore, we show a rapid, coordinated depolarization of the membrane potential component of the proton-motive force throughout the cell in response to spatially controlled uncoupling of the cell interior. We propose that membrane potential conduction via the mitochondrial reticulum is the dominant pathway for skeletal muscle energy distribution.

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Acknowledgements

We would like to thank E. Tyler for his assistance with image rendering, J. Taraska for providing experimental advice, T. Karpova and the NCI Core Fluorescence Imaging Facility for access to a microscope with a 355-nm laser, and P. Kellman for assistance with image analysis. We would also like to thank S. Caldwell and R. Hartley for providing us with MitoPhotoDNP. This study was supported by intramural funding of the Division of Intramural Research, National Heart, Lung, and Blood Institute and the Center for Cancer Research, National Cancer Institute.

Author information

Author notes

    • Brian Glancy
    •  & Lisa M. Hartnell

    These authors contributed equally to this work.

Affiliations

  1. National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Brian Glancy
    • , Daniela Malide
    • , Zu-Xi Yu
    • , Christian A. Combs
    • , Patricia S. Connelly
    •  & Robert S. Balaban
  2. National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Lisa M. Hartnell
    •  & Sriram Subramaniam

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Contributions

R.S.B., S.S., B.G. and L.M.H. designed and L.M.H performed the FIB-SEM experiments. R.S.B. and B.G. designed and B.G. performed the MPM experiments. R.S.B., B.G., Z.-X.Y. and D.M. designed and B.G., Z.-X.Y. and D.M. performed the dual immunolabelling experiments. P.S.C. performed the TEM imaging. R.S.B. and B.G. performed the segmentations and analysed the data. R.S.B., C.A.C. and B.G. designed and performed the isolated muscle fibre experiments.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Robert S. Balaban.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains the full gel image of data shown in Extended Data Figure 5a (highlighted by red box) and full western blot image of data shown in Extended Data Figure 5a (highlighted by red box).

Videos

  1. 1.

    Begins with a 3D FIB/SEM image stack of mouse Tibialis anterior muscle.

    Longitudinal images are shown and time represents sequential images moving deeper into the fiber (15 nm steps). These are the data from which Figures 1 and 2a-f were created. Then the same image stack after automated mitochondrial segmentation is shown followed by a 3D rendering of the segmented mitochondria. Close up views of IBM projections from PVM as well as the fiber interior are also shown, respectively. Raw image stack is available as Supplementary Dataset 1 at http://labalaban.nhlbi.nih.gov/files/SuppDataset.tif.

  2. 2.

    3D rendering of the data in Figure 2a.

    3D rendering of the data in Figure 2a.

  3. 3.

    Longitudinal MPM image stack for the data shown Extended Data Figure 2.

    Time represents sequential images moving deeper into the muscle fiber (100 nm steps).

  4. 4.

    Cross-sectional view of the MPM image stack shown in Extended Data Figure 2.

    Time represents sequential images moving down the barrel of the fiber (100 nm steps).

  5. 5.

    3D rendering of the MPM muscle volume shown in Extended Data Figure 2.

    3D rendering of the MPM muscle volume shown in Extended Data Figure 2.

  6. 6.

    Longitudinal STED image stack of an isolated muscle cell stained with mitochondrial membrane potential probe TMRM.

    Time represents sequential images moving deeper into the muscle fiber (219 nm steps). Images shown are raw data.

  7. 7.

    360° rotation of a 3D rendering of the ratiometric determination of the spatial distribution of mitochondrial Complexes IV and V as shown in Figure 3.

    360° rotation of a 3D rendering of the ratiometric determination of the spatial distribution of mitochondrial Complexes IV and V as shown in Figure 3.

  8. 8.

    Individual channel and merged images of the raw data for the muscle section dual immunostaining shown in Figure 3.

    Upper left (green) – Complex IV, upper right (red) – Complex V, lower left (blue) – nuclei, lower right – merged images.

  9. 9.

    Timecourse loop of confocal microscopy images of the TMRM signal in a live, isolated muscle fiber before and after UV light induced release of MitoPhotoDNP in the cell interior.

    Timecourse loop of confocal microscopy images of the TMRM signal in a live, isolated muscle fiber before and after UV light induced release of MitoPhotoDNP in the cell interior.

  10. 10.

    Timecourse loop of confocal microscopy images of the TMRM signal in a live, isolated muscle fiber before and after UV light exposure in the cell interior of a fiber without MitoPhotoDNP.

    Timecourse loop of confocal microscopy images of the TMRM signal in a live, isolated muscle fiber before and after UV light exposure in the cell interior of a fiber without MitoPhotoDNP.

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DOI

https://doi.org/10.1038/nature14614

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