Analysis of ER-mitochondria contacts using correlative fluorescence microscopy and soft X-ray tomography of mammalian cells

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TitleAnalysis of ER-mitochondria contacts using correlative fluorescence microscopy and soft X-ray tomography of mammalian cells
Publication TypeJournal Article
Year of Publication2015
AuthorsElgass KD, Smith EA, Le Gros MA, Larabell CA, Ryan MT
Journal TitleJ Cell Sci
Volume128
Pages2795-804
Journal DateAug 01
ISBN Number1477-9137 (Electronic)0021-9533 (Linking)
Accession Number26101352
KeywordsAnimals, Apoptosis/physiology, Cell Line, Tumor, Cercopithecus aethiops, Confocal microscopy, Correlative imaging, COS Cells, Dynamins, Endoplasmic reticulum, Endoplasmic Reticulum/*metabolism, Imaging, Three-Dimensional, Membrane Proteins/*genetics, Mice, Microscopy imaging, Microscopy, Confocal, Microscopy, Fluorescence, Mitochondria, Mitochondria/*metabolism, Mitochondrial Dynamics/*physiology, Mitochondrial fission, Mitochondrial Membranes/*metabolism, Mitochondrial Proteins/*genetics, Receptors, Cytoplasmic and Nuclear/*genetics, soft x-ray tomography, Tomography, X-Ray Computed
Abstract

Mitochondrial fission is important for organelle transport, quality control and apoptosis. Changes to the fission process can result in a wide variety of neurological diseases. In mammals, mitochondrial fission is executed by the GTPase dynamin-related protein 1 (Drp1; encoded by DNM1L), which oligomerizes around mitochondria and constricts the organelle. The mitochondrial outer membrane proteins Mff, MiD49 (encoded by MIEF2) and MiD51 (encoded by MIEF1) are involved in mitochondrial fission by recruiting Drp1 from the cytosol to the organelle surface. In addition, endoplasmic reticulum (ER) tubules have been shown to wrap around and constrict mitochondria before a fission event. Up to now, the presence of MiD49 and MiD51 at ER-mitochondrial division foci has not been established. Here, we combine confocal live-cell imaging with correlative cryogenic fluorescence microscopy and soft x-ray tomography to link MiD49 and MiD51 to the involvement of the ER in mitochondrial fission. We gain further insight into this complex process and characterize the 3D structure of ER-mitochondria contact sites.

URLhttps://www.ncbi.nlm.nih.gov/pubmed/26101352http://jcs.biologists.org/content/joces/128/15/2795.full.pdf