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Accumulation of anchored proteins forms membrane diffusion barriers during neuronal polarization

Nature Cell Biology volume 5, pages 626–632 (2003)Cite this article

Abstract

The formation and maintenance of polarized distributions of membrane proteins in the cell membrane are key to the function of polarized cells. In polarized neurons, various membrane proteins are localized to the somatodendritic domain or the axon. Neurons control polarized delivery of membrane proteins to each domain, and in addition, they must also block diffusional mixing of proteins between these domains. However, the presence of a diffusion barrier in the cell membrane of the axonal initial segment (IS), which separates these two domains, has been controversial: it is difficult to conceive barrier mechanisms by which an even diffusion of phospholipids could be blocked. Here, by observing the dynamics of individual phospholipid molecules in the plasma membrane of developing hippocampal neurons in culture, we found that their diffusion was blocked in the IS membrane. We also found that the diffusion barrier is formed in neurons 7–10 days after birth through the accumulation of various transmembrane proteins that are anchored to the dense actin-based membrane skeleton meshes being formed under the IS membrane. We conclude that various membrane proteins anchored to the dense membrane skeleton function as rows of pickets, which even stop the overall diffusion of phospholipids, and may represent a universal mechanism for formation of diffusion barriers in the cell membrane.

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Figure 1: Immobilisation of DOPE in the IS membrane of older neurons.
Figure 2: DOPE diffusion is suppressed only in the IS of older neurons.
Figure 3: Developmental barrier formation.
Figure 4: The barrier mechanism of the IS membrane.

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Acknowledgements

We thank M. Aihara and M. Nozaki for help with culturing neurons. We also thank V. Bennett and his colleagues, M. Poo, B. Winckler, T. Kobayashi, and Y. Tsuchimoto for helpful discussions.

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Author notes

  1. Mitsuhiro Nakamura

    Present address: Department of Applied Physics and Chemistry, The University of Electro-Communications, Chofu, Tokyo, 182-8585, Japan

Authors and Affiliations

  1. Kusumi Membrane Organizer Project, Exploratory Research for Advancement of Technology Organization (ERATO), JST, Nagoya, 460-0012, Japan

    Chieko Nakada, Kenneth Ritchie, Yoko Hotta, Ryota Iino, Takahiro Fujiwara & Akihiro Kusumi

  2. Department of Biological Science and Institute for Advanced Research, Nagoya University, Nagoya, 464-8602, Japan

    Kenneth Ritchie, Rinshi S. Kasai & Akihiro Kusumi

  3. Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, 464-8601, Japan

    Yuichi Oba & Mitsuhiro Nakamura

  4. Laboratory for Memory and Learning, Brain Science Institute, RIKEN, Hirosawa 2-1, Wako, 351-0198, Saitama, Japan

    Kazuhiko Yamaguchi

  5. Yoshida ATP System Project, ERATO, JST, Nagatsuta 5800-3, Midori-ku, 226-0026, Yokohama, Japan

    Ryota Iino

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Correspondence to Akihiro Kusumi.

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Nakada, C., Ritchie, K., Oba, Y. et al. Accumulation of anchored proteins forms membrane diffusion barriers during neuronal polarization. Nat Cell Biol 5, 626–632 (2003). https://doi.org/10.1038/ncb1009

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