Electron tomography reveals unbranched networks of actin filaments in lamellipodia

Abstract

Electron tomography provides unprecedented three-dimensional images of lamellipodial actin in frozen, ‘live’ cells. The vast majority of actin filaments at the leading edge are unbranched, requiring a re-examination of the role of Arp2/3-mediated branching in vivo. Eukaryotic cells can initiate movement using the forces exerted by polymerizing actin filaments to extend lamellipodial and filopodial protrusions. In the current model, actin filaments in lamellipodia are organized in a branched, dendritic network. We applied electron tomography to vitreously frozen 'live' cells, fixed cells and cytoskeletons, embedded in vitreous ice or in deep-negative stain. In lamellipodia from four cell types, including rapidly migrating fish keratocytes, we found that actin filaments are almost exclusively unbranched. The vast majority of apparent filament junctions proved to be overlapping filaments, rather than branched end-to-side junctions. Analysis of the tomograms revealed that actin filaments terminate at the membrane interface within a zone several hundred nanometres wide at the lamellipodium front, and yielded the first direct measurements of filament densities. Actin filament pairs were also identified as lamellipodium components and bundle precursors. These data provide a new structural basis for understanding actin-driven protrusion during cell migration.

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Urban, E., Jacob, S., Nemethova, M. et al. Electron tomography reveals unbranched networks of actin filaments in lamellipodia. Nat Cell Biol 12, 429–435 (2010). https://doi.org/10.1038/ncb2044

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