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Whole Cell Cryo-Electron Tomography Suggests Mitochondria Divide by Budding

Published online by Cambridge University Press:  28 May 2014

Guo-Bin Hu
Guo-Bin Hu*
Affiliation:
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA
*
*Corresponding author. [email protected]
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Abstract

Eukaryotes rely on mitochondrial division to guarantee that each new generation of cells acquires an adequate number of mitochondria. Mitochondrial division has long been thought to occur by binary fission and, more recently, evidence has supported the idea that binary fission is mediated by dynamin-related protein (Drp1) and the endoplasmic reticulum. However, studies to date have depended on fluorescence microscopy and conventional electron microscopy. Here, we utilize whole cell cryo-electron tomography to visualize mitochondrial division in frozen hydrated intact HeLa cells. We observe a large number of relatively small mitochondria protruding from and connected to large mitochondria or mitochondrial networks. Therefore, this study provides evidence that mitochondria divide by budding.

Keywords

cryo-EMcryo-electron tomographymitochondrionmitochondrial divisionmitochondrial buddingmitochondrial dynamics
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 20 , Issue 4 , August 2014 , pp. 1180 - 1187
Copyright
© Microscopy Society of America 2014 

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References

Beech, P.L., Nheu, T., Schultz, T., Herbert, S., Lithgow, T., Gilson, P.R. & McFadden, G.I. (2000). Mitochondrial FtsZ in a chromophyte alga. Science 287, 1276–1279.Google Scholar
Benard, G. & Karbowski, M. (2009). Mitochondrial fusion and division: Regulation and role in cell viability. Semin Cell Dev Biol 20, 365–374.CrossRefGoogle ScholarPubMed
Blackstone, C. & Chang, C.R. (2011). Mitochondrial unite to survive. Nat Cell Biol 13, 521–522.Google Scholar
Chan, D.C. (2006). Mitochondrial fusion and fission in mammals. Annu Rev Cell Dev Biol 27, 79–99.Google Scholar
Davison, M.T. & Garland, P.B. (1977). Structure of mitochondria and vacuoles of Candida utilis and Schizosaccharomyces pombe studied by electron microscopy of serial thin sections and model building. J Gen Microbiol 98, 147–153.CrossRefGoogle ScholarPubMed
Fawcett, D.W. (1981). The C ell , 2nd ed. Philadelphia, PA, USA: Saunders Press. pp. 430–437.Google Scholar
Frank, J. (2006). Electron Tomography: Methods for Three-Dimensional Visualization of Structures in the Cell, 2nd ed. New York, NY, USA: Springer.Google Scholar
Frey, T.G. & Mannella, C.A. (2000). The internal structure of mitochondria. Trends Biochem Sci 25, 319–324.Google Scholar
Fridman, K., Mader, A., Awerger, M., Elia, N. & Medalia, O. (2012). Advances in tomography: Probing the molecular architecture of cells. Nat Rev Mol Cell Biol 13, 736–742.CrossRefGoogle ScholarPubMed
Friedman, J.R., Lackner, L.L., West, M., DiBenedetto, J.R., Nunnari, J. & Voeltz, G.K. (2011). ER tubules mark sites of mitochondrial division. Science 334, 358–362.Google Scholar
Gray, M.W., Burger, G. & Lang, B.F. (1999). Mitochondrial evolution. Science 283, 1476–1481.Google Scholar
Haust, M.D. (1968). Mitochondrial budding and morphogenesis of cytoplasmic vacuoles in hepatocytes of children with the Hurler syndrome and Sanfilippo disease. Exp Mol Pathol 9, 242–257.Google Scholar
Hirsch, P. (1974). Budding bacteria. Annu Rev Microbiol 28, 391–444.CrossRefGoogle ScholarPubMed
Hom, J.R., Quintanilla, R.A., Hoffman, D.L., de Mesy Bentley, K.L., Molkentin, J.D., Sheu, S.S. & Porter, G.A. Jr. (2011). The permeability transition pore controls cardiac mitochondrial maturation and myocyte differentiation. Dev Cell 146, 798–812.Google Scholar
Hoppins, S., Lackner, L. & Nunnari, J. (2007). The machines that divide and fuse mitochondria. Annu Rev Biochem 76, 751–780.Google Scholar
Ishihara, N., Nomura, M., Jofuku, A., Kato, H., Suzuki, S.O., Masuda, K., Otera, H., Nakanishi, Y., Nonaka, I., Goto, Y., Taguchi, N., Morinaga, H., Maeda, M., Takayanagi, R., Yokota, S. & Mihara, K. (2009). Mitochondrial fission factor Drp1 is essential for embryonic development and synapse formation in mice. Nat Cell Biol 11, 958–966.CrossRefGoogle ScholarPubMed
Kashatus, D.F., Lim, K.H., Brady, D.C., Pershing, N.L. & Cox, A.D. (2011). RALA and RALBP1 regulate mitochondrial fission at mitosis. Nat Cell Biol 13, 1108–1115.Google Scholar
Kremer, J.R., Mastronarde, D.N. & McIntosh, J.R. (1996). Computer visualization of three-dimensional image data using IMOD. J Struct Biol 116, 71–76.Google Scholar
Larsen, W.J. (1970). Genesis of mitochondria in insect fat body. J Cell Biol 47, 373–383.Google Scholar
Lewis, M.R. & Lewis, W.H. (1914). Mitochondrial in tissue culture. Science 39, 330–333.CrossRefGoogle ScholarPubMed
Lo, Y.S., Hsiao, L.J., Jane, W.N., Cheng, N. & Dai, H. (2011). Ultrastructural studies on actin-like filament in mung bean mitochondria and its potential functional significance. Bot Stud 52, 445–454.Google Scholar
Matarrese, P., Tinari, A., Mormone, E., Bianco, G.A., Toscano, M.A., Ascione, B., Rabinovich, G.A. & Malorni, W. (2005). Galectin-1 sensitizes resting human T lymphocytes to Fas (CD95)-mediated cell death via mitochondrial hyperpolarization, budding, and fission. J Biol Chem 280, 6969–6985.Google Scholar
Medalia, O., Weber, I., Frangakis, A.S., Nicastro, D., Gerisch, G. & Baumeister, W. (2002). Macrolmolecular architecture in eukaryotic cells visualized by cryoelectron tomography. Science 298, 1209–1213.Google Scholar
Meeusen, S., McCaffery, J.M. & Nunnari, J. (2004). Mitochondrial fusion intermediates revealed in vitro. Science 305, 1747–1752.CrossRefGoogle ScholarPubMed
Montessuit, S., Somasekharan, S.P., Terrones, O., Lucken-Ardjomande, S., Herzig, S., Schwarzenbacher, R., Manstein, D.J., Bossy-Wetzel, E., Basañez, G., Meda, P. & Martinou, J.C. (2010). Membrane remodeling induced by the dynamin-related protein Drp1 stimulates Bax oligomerization. Cell 142, 889–901.Google Scholar
Neuspiel, M., Schauss, A.C., Braschi, E., Zunino, R., Rippstein, P., Rachubinski, R.A., Andrade-Navarro, M.A. & McBride, H.M. (2008). Cargo-selected transport from the mitochondria to peroxisomes is mediated by vesicular carriers. Curr Biol 18, 102–108.Google Scholar
Nicastro, D., Frangakis, A.S., Typke, D. & Baumeister, W. (2000). Cryo-electron tomography of neurospora mitochondria. J Struct Biol 129, 48–56.Google Scholar
Otera, H., Wang, C., Cleland, M.M., Setoguchi, K., Yokota, S., Youle, R.J. & Mihara, K. (2010). Mff is an essential factor for mitochondrial recruitment of Drp1 during mitochondrial fission in mammalian cells. J Cell Biol 191, 1141–1158.CrossRefGoogle ScholarPubMed
Palmer, C.S., Osellame, L.D., Laine, D., Koutsopoulos, O.S., Frazier, A.E. & Ryan, M.T. (2011). MiD49 and MiD51, new components of the mitochondrial fission machinery. EBMO Rep 12, 565–573.Google Scholar
Perkins, G., Bossy-Wetzel, E. & Ellisman, M.H. (2009). New insights into mitochondrial structure during cell death. Exp Neurol 218, 183–192.Google Scholar
Pesce, G. & Walter, P. (2007). Video 14.2 mitochondrial fission and fusion. In Molecular Biology of the Cell, 5th ed. Alberts B. et al. (Eds.), Video 14.2. Garland Science.Google Scholar
Pettersen, E.F., Goddard, T.D., Huang, C.C., Couch, G.S., Greenblatt, D.M., Meng, E.C. & Ferrin, T.E. (2004). UCSF Chimera—a visualization system for exploratory research and analysis. J Comput Chem 25, 1605–1612.CrossRefGoogle ScholarPubMed
Pilhofer, M., Ladinsky, M., McDowall, A.W., Petroni, G. & Jensen, G.J. (2011). Microtubules in bacteria: Ancient tubulins build a five-protofilament homolog of the eukaryotic cytoskeleton. PloS Biol 9(12), e100123.CrossRefGoogle ScholarPubMed
Shim, S.H., Xia, C., Zhong, G., Babcock, H.P., Vaughan, J.C., Huang, B., Wang, X., Cheng, X., Bi, G.Q. & Zhuang, X. (2012). Super-resolution fluorescent imaging of organelles in live cells with photoswitchable membrane probes. Proc Natl Acad Sci USA 109, 13978–13983.Google Scholar
Smirnova, E., Griparic, L., Shurland, D.L. & van der Bliek, A.M. (2001). Dynamin-related protein Drp1 is required for mitochondrial division in mammalian cells. Mol Biol Cell 12, 2245–2256.Google Scholar
Soubannier, V., McLelland, G.L., Zunino, R., Braschi, E., Rippstein, P., Fon, E.A. & McBride, H.M. (2012 a). A vesicular transport pathway shuttle cargo from mitochondria to lysosomes. Curr Biol 22, 135–141.Google Scholar
Soubannier, V., Rippstein, P., Kaufman, B.A., Shoubridge, E.A. & McBride, H.M. (2012 b). Reconstitution of mitochondria derived vesicle formation demonstrates selective enrichment of oxidized cargo. PloS One 7, e52830.Google Scholar
Steven, A.C. & Aebi, U. (2003). The next ice age, cryo-electron tomography of intact cells. Trends Cell Biol 13, 107–110.CrossRefGoogle ScholarPubMed
Tandler, B. & Hoppel, C.L. (1973). Division of giant mitochondria during recovery from cuprizone intoxication. J Cell Biol 56, 266–272.CrossRefGoogle ScholarPubMed
Thévenaz, P. & Unser, M. (2007). User-friendly semiautomated assembly of accurate image mosaics in microscopy. Microsc Res and Tech 70, 135–146.Google Scholar
van Driel, L.F., Valentijn, J.A., Valentijn, K.M., Koning, R.I. & Koster, A.J. (2009). Tools for correlative cryo-fluorescence microscopy and cryo-electron tomography applied to whole mitochondria in human endothelial cells. Eur J Cell Biol 88, 669–684.Google Scholar
Westermann, B. (2010). Mitochondrial fusion and fission in cell life and death. Nat Rev Mol Cell Biol 11, 872–884.CrossRefGoogle ScholarPubMed
Winkler, H. & Taylor, K.A. (2006). Accurate marker-free alignment with simultaneous geometry determination and reconstruction of tilt series in electron tomography. Ultramicroscopy 106, 240–254.Google Scholar
Youle, R.J. & Narendra, D.P. (2011). Mechanism of mitophagy. Nat Rev Mol Cell Biol 12, 9–14.Google Scholar
Youle, R.J. & van der Bliek, A.M. (2012). Mitochondrial fission, fusion, and stress. Science 337, 1062–1065.CrossRefGoogle ScholarPubMed
Zhao, J., Liu, T., Jin, S., Wang, X., Qu, M., Uhlén, P., Tomilin, N., Shupliakov, O., Lendahl, U. & Nistér, M. (2011). Human MIEF1 recruits Drp1 to mitochondrial outer membranes and promotes mitochondrial fusion rather than fission. EBMO J 30, 2762–2778.Google Scholar

Hu Supplementary Material

Movie S1, a round-trip slice-wise show of the 3D map associated with Fig. 2A and D.

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Video 9 MB

Hu Supplementary Material

Movie S2, a round-trip slice-wise display of the 3D map associated with Fig. 2B and E.

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Video 10.3 MB

Hu Supplementary Material

Movie S3, a round-trip slice-wise display of the 3D map associated with Fig. 2C and F.

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Video 6.4 MB
Supplementary material: File

Hu Supplementary Material

Figures S1-S4

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