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Actin restricts FcɛRI diffusion and facilitates antigen-induced receptor immobilization

Nature Cell Biology volume 10, pages 955–963 (2008)Cite this article

Abstract

The actin cytoskeleton has been implicated in restricting diffusion of plasma membrane components. Here, simultaneous observations of quantum dot-labelled FcɛRI motion and GFP-tagged actin dynamics provide direct evidence that actin filament bundles define micron-sized domains that confine mobile receptors. Dynamic reorganization of actin structures occurs over seconds, making the location and dimensions of actin-defined domains time-dependent. Multiple FcɛRI often maintain extended close proximity without detectable correlated motion, suggesting that they are co-confined within membrane domains. FcɛRI signalling is activated by crosslinking with multivalent antigen. We show that receptors become immobilized within seconds of crosslinking. Disruption of the actin cytoskeleton results in delayed immobilization kinetics and increased diffusion of crosslinked clusters. These results implicate actin in membrane partitioning that not only restricts diffusion of membrane proteins, but also dynamically influences their long-range mobility, sequestration and response to ligand binding.

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Figure 1: QD-IgE serves as a non-perturbing probe of FcɛRI diffusion.
Figure 2: FcɛRI are co-confined.
Figure 3: Motion of QD-IgE–FcɛRI is consistent with co-confinement not attraction.
Figure 4: Actin defines regions of FcɛRI motion in the membrane.
Figure 5: Effect of actin proximity on FcɛRI trajectories.
Figure 6: Actin facilitates crosslink-induced immobilization of FcɛRI.

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Acknowledgements

This work was supported by NIH grants R01 GM49814, R01 AI051575 and P20 GM 67594, the Oxnard Foundation, ACS IRG 192 and by the Sandia LDRD and SURP programs. Sandia is a multi-program laboratory operated by Sandia Corporation, a Lockheed Martin Company, for the United States Department of Energy under contract DE-AC04-94AL85000. N.A. was supported by NSF IGERT DGE-0549500 and the UNM-SOM MD/PhD Program. We thank Sheli Ryan for assistance with cell culture, Jun Zhang for EM image analysis, and Jonas Anderson and Bernd Rieger for assistance with image processing. The UNM Cancer Center Fluorescence Microscopy Facility received support from NIH grants S10 RR14668, S10 RR19287, S10 RR016918, P20 RR11830 and P30 CA118100 and from NSF grant MCB9982161. Electron micrographs were generated in the University of New Mexico Electron Microscopy Facility, which received support from NIH grants P20 GM067594, S10 RRI5734 and RR022493.

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  1. Nicholas L. Andrews and Keith A. Lidke: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology and Cancer Research and Treatment Center, University of New Mexico, Albuquerque, 87131, New Mexico, USA

    Nicholas L. Andrews, Janet R. Pfeiffer, Bridget S. Wilson, Janet M. Oliver & Diane S. Lidke

  2. Department of Physics and Astronomy, University of New Mexico, Albuquerque, 87131, New Mexico, USA

    Keith A. Lidke

  3. Sandia National Laboratories, Albuquerque, 87185-0351, New Mexico, USA

    Keith A. Lidke & Alan R. Burns

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Andrews, N., Lidke, K., Pfeiffer, J. et al. Actin restricts FcɛRI diffusion and facilitates antigen-induced receptor immobilization. Nat Cell Biol 10, 955–963 (2008). https://doi.org/10.1038/ncb1755

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