Abstract

During mitochondrial fission, key molecular and cellular factors assemble on the outer mitochondrial membrane, where they coordinate to generate constriction. Constriction sites can eventually divide, or reverse upon disassembly of the machinery. However, a role for membrane tension in mitochondrial fission, although speculated, has remained undefined. We captured the dynamics of constricting mitochondria in mammalian cells using live-cell structured illumination microscopy (SIM). By analyzing the diameters of tubules that emerge from mitochondria and implementing a fluorescence lifetime-based mitochondrial membrane tension sensor, we discovered that mitochondria are indeed under tension. Under perturbations that reduce mitochondrial tension, constrictions initiate at the same rate, but are less likely to divide. We propose a model based on our estimates of mitochondrial membrane tension and bending energy in living cells which accounts for the observed probability distribution for mitochondrial constrictions to divide.

Highlights

  • Mitochondria are highly dynamic organelles, transported through the cytoplasm along cytoskeletal networks while they change in size and shape (Nunnari et al, 1997; Youle and van der Bliek, 2012)

  • Constriction by the division machinery does not ensure fission We performed live-cell structured illumination microscopy (SIM) imaging of COS-7 cells transiently transfected with Drp1mCherry and GFP targeted to the matrix by the mitochondrial targeting sequence from subunit VIII of human cytochrome c oxidase

  • Since our SIM imaging was limited to 2 colors, we performed fast (1 Hz), 3-color live-cell confocal imaging of mitochondria and Drp1, with either dynamin 2 protein (Dyn2) or the endoplasmic reticulum (ER) (Figures 1E–1L)

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Summary

Introduction

Mitochondria are highly dynamic organelles, transported through the cytoplasm along cytoskeletal networks while they change in size and shape (Nunnari et al, 1997; Youle and van der Bliek, 2012). The fission site is marked by a pre-constriction defined by contact with endoplasmic reticulum (ER) tubules (Friedman et al, 2011) and deformed by targeted actin polymerization (Ji et al, 2015; Korobova et al, 2013; Manor et al, 2015) and myosin II contraction (Hatch et al, 2014; Korobova et al, 2014; Yang and Svitkina, 2019) Surface receptors such as MiD49/51 (Palmer et al, 2011) or Mff (Gandre-Babbe and van der Bliek, 2008; Otera et al, 2010) accumulate at the preconstriction site and recruit dynamin-related protein (Drp1) (Labrousse et al, 1999; Smirnova et al, 2001). This underlines the fundamentally mechanical nature of membrane fission processes

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