Journal
JOURNAL OF CELL BIOLOGY
Volume 218, Issue 1, Pages 83-96Publisher
ROCKEFELLER UNIV PRESS
DOI: 10.1083/jcb.201809107
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Funding
- G. Harold and Leila Y. Mathers Foundation
- Wellcome Trust [095927/A/11/Z, 203285/B/16/Z]
- National Institutes of Health [T32 GM100884, T32 GM007499, R01 GM118486, P30 DK045735, S10 OD020142, T32 GM008283]
- Yale University
- Gruber Foundation
- Austrian Marshall Plan Foundation
- NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES [P30DK045735] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [T32GM007499, T32GM100884, R01GM118486, T32GM008283] Funding Source: NIH RePORTER
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The endoplasmic reticulum (ER) is composed of interconnected membrane sheets and tubules. Superresolution microscopy recently revealed densely packed, rapidly moving ER tubules mistaken for sheets by conventional light microscopy, highlighting the importance of revisiting classical views of ER structure with high spatiotemporal resolution in living cells. In this study, we use live-cell stimulated emission depletion (STED) microscopy to survey the architecture of the ER at 50-nm resolution. We determine the nanoscale dimensions of ER tubules and sheets for the first time in living cells. We demonstrate that ER sheets contain highly dynamic, subdiffraction-sized holes, which we call nanoholes, that coexist with uniform sheet regions. Reticulon family members localize to curved edges of holes within sheets and are required for their formation. The luminal tether Climp63 and microtubule cytoskeleton modulate their nanoscale dynamics and organization. Thus, by providing the first quantitative analysis of ER membrane structure and dynamics at the nanoscale, our work reveals that the ER in living cells is not limited to uniform sheets and tubules; instead, we suggest the ER contains a continuum of membrane structures that includes dynamic nanoholes in sheets as well as clustered tubules.
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