Imaging axonal transport of mitochondria in vivo (original) (raw)

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Acknowledgements

We thank P. Glass, K. Mahoney, D. Malkowski and S. Haddad for administrative support and animal husbandry, and J. Sanes for valuable suggestions. We thank C. McCann and J. Morgan for advice on anatomical analysis, and L. Godinho for critically reading the manuscript. We also thank the employees of electron microscopy and microinjection services at the Jackson Laboratory. This work was supported by grants from the Dana Foundation (T.M. and M.K.), the Emmy Noether program of the Deutsche Forschungsgemeinschaft (M.K.), the Alexander-von-Humboldt foundation (T.M.), the “Verein Therapieforschung für MS-Kranke e.V.” (M.K.), the European Molecular Biology Organization (F.M.B.), the ALS Association (R.W.B.) and the US National Institutes of Health (J.W.L.).

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Author notes

  1. Thomas Misgeld and Martin Kerschensteiner: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular and Cellular Biology, Harvard University, 7 Divinity Avenue, Cambridge, 02138, Massachusetts, USA
    Thomas Misgeld, Martin Kerschensteiner, Florence M Bareyre & Jeff W Lichtman
  2. Institute of Neurosciences, Technical University Munich, Munich, 80802, Germany
    Thomas Misgeld
  3. Institute of Clinical Neuroimmunology, Ludwig-Maximilians University, Munich, 81377, Germany
    Martin Kerschensteiner & Florence M Bareyre
  4. The Jackson Laboratory, Bar Harbor, 04609, Maine, USA
    Robert W Burgess

Authors

  1. Thomas Misgeld
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  2. Martin Kerschensteiner
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  3. Florence M Bareyre
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  4. Robert W Burgess
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  5. Jeff W Lichtman
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Corresponding authors

Correspondence toThomas Misgeld or Jeff W Lichtman.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Figures and Text

Supplementary Figures 1–4, Supplementary Methods. (PDF 2503 kb)

Supplementary Video 1

Time-lapse imaging of mitochondrial transport. The movie shows two axons in an intercostal nerve in the triangularis sterni explant. A node of Ranvier (NR) is present in the upper axon. The majority of mitochondria are immobile, but a fraction moves quickly (1-1.5 μm / sec) in anterograde (left to right) and retrograde directions. At nodes of Ranvier transported mitochondria often slow down and sometimes stall, especially those transported in the retrograde direction (anterograde velocity crossing a node 0.73 ± 0.06 μm per sec, n = 25; retrograde 0.57 ± 0.04 μm per sec, n = 25). Time is indicated in min: sec. (MOV 2438 kb)

Supplementary Video 2

Different mitochondrial populations in living axons in an acute nerve-muscle explant. The movie shows the intercostal axon represented in Figure 1d; boxes indicate the mitochondria highlighted there. The lower panel shows the same axon after image processing and intensity inversion. Time is indicated in min: sec. (MOV 2426 kb)

Supplementary Video 3

Mitochondrial transport at an axonal branch point. Trifurcation in an intercostal axon in the triangularis sterni explant imaged for 45 minutes (thy1-mitoCFP-S x thy1-YFP-16). Numerous mitochondria enter the three different branches (# 1 - 3). A time-averaged version of this movie is shown in Supplementary Figure 3b. Note deceleration of many mitochondria as they transit the branch point. Time is indicated in min: sec. (MOV 2488 kb)

Supplementary Video 4

Mitochondrial dynamics in growing and retreating sprouts. Two axon sprouts in an acute nerve-muscle explants prepared one week after axon transection in the living animal(thy1-mitoCFP-S x thy1-YFP-16; their location is indicated in Figure 2a). Over about 2 hours, the upper sprout retreats, while the lower one extends a growth cone. Mitochondria can be seen advancing into axonal varicosities along the stem of the growing sprout. Time is indicated in min: sec. CFP shown in cyan; YFP in yellow. (MOV 2124 kb)

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Misgeld, T., Kerschensteiner, M., Bareyre, F. et al. Imaging axonal transport of mitochondria in vivo.Nat Methods 4, 559–561 (2007). https://doi.org/10.1038/nmeth1055

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