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A diffusion barrier maintains distribution of membrane proteins in polarized neurons

Abstract

The asymmetric distribution of proteins to distinct domains in the plasma membrane is crucial to the function of many polarized cells. In epithelia, distinct apical and basolateral surfaces are maintained by tight junctions that prevent diffusion of proteins and lipids between the two domains1. Polarized neurons maintain axonal and somatodendritic plasma membrane domains without an obvious physical barrier. Indeed, the artificial lipid DiI encounters no diffusion barrier at the presumptive domain boundary, the axon hillock2. By measuring the lateral mobility of membrane proteins using optical tweezers, we show here that some membrane proteins exhibit markedly reduced mobility in the initial segment of the axon. Disruption of F-actin and low levels of dimethyl sulphoxide (DMSO) abolish this diffusion barrier and lead to redistribution of membrane markers that had previously been polarized. Immobilization in the initial segment may reflect, at least in part, differential tethering to cytoskeletal components. Therefore, the ability to maintain a polarized distribution of membrane proteins depends on a specialized domain at the initial segment of the axon, which restricts lateral mobility and serves as a new type of diffusion barrier that acts in the absence of cell–cell contact.

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Figure 1: The axonal initial segment compartmentalizes axonal and somatodendritic domains.
Figure 2: Bead mobility is restricted in the axon initial segment in control but not DMSO- or LAT-treated cells.
Figure 3: DMSO and LAT-B treatment lead to randomization of axonal NgCAM.
Figure 4: L1 and NgCAM are part of a DMSO- and LAT-sensitive CHAPS-resistant complex at the initial segment.

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References

  1. Mellman, I. Molecular sorting of membrane proteins in polarized and non-polarized cells. Cold Spring Harb. Symp. Quant. Biol. 60, 745–752 (1996).

    Article  Google Scholar 

  2. Winckler, B. & Poo, M.-m. No diffusion barrier at axon hillock. Nature 379, 213 (1996).

    Article  ADS  CAS  Google Scholar 

  3. Vogt, L. et al. Continuous renewal of the axonal pathway sensor apparatus by insertion of new sensor molecules into the growth cone membrane. Curr. Biol. 6, 1153–1158 (1996).

    Article  CAS  Google Scholar 

  4. Dotti, C. G., Parton, R. G. & Simons, K. Polarized sorting of glypiated proteins in hippocampal neurons. Nature 349, 156–161 (1991).

    Article  ADS  Google Scholar 

  5. Kobayashi, T., Storrie, B., Simons, K. & Dotti, C. G. A functional barrier to movements of lipids in polarized neurons. Nature 359, 647–650 (1992).

    Article  ADS  CAS  Google Scholar 

  6. Kusumi, A., Sako, Y., Fujiwara, T. & Tomishige, M. Application of laser tweezers to studies of the fences and tethers of the membrane skeleton that regulate the movements of plasma membrane proteins. Methods Cell Biol. 55, 173–194 (1998).

    Article  CAS  Google Scholar 

  7. Dai, J. & Sheetz, M. P. Mechanical properties of neuronal growth cone membranes studied by tether formation with laser optical tweezers. Biophys. J. 68, 988–996 (1995).

    Article  ADS  CAS  Google Scholar 

  8. Lamaze, C., Fijimoto, L. M., Yin, H. L. & Schmid, S. L. The actin cytoskeleton is required for receptor-mediated endocytosis in mammalian cells. J. Biol. Chem. 272, 20332 (1997).

    Article  CAS  Google Scholar 

  9. Kusumi, A., Sako, Y. & Yamamoto, M. Confined lateral diffusion of membrane receptors as studied by single particle tracking (nanovid microscopy). Effects of calcium-induced differentiation in cultured epithelial cells. Biophys. J. 65, 2021–2040 (1993).

    Article  ADS  CAS  Google Scholar 

  10. Simons, K. & Ikonen, E. functional rafts in cell membranes. Nature 387, 569–572 (1997).

    Article  ADS  CAS  Google Scholar 

  11. Peters, A., Proskauer, C. C. & Kaiserman-Abramof, I. R. The small pyramidal neuron of the rat cerebral cortex: the axon hillock and initial segment. J. Cell Biol. 39, 604–619 (1968).

    Article  CAS  Google Scholar 

  12. Bartlett, W. P. & Banker, G. A. An electron microscopic study of the development of axons and dendrites by hippocampal neurons in culture. I. Cells which develop without intercellular contacts. J. Neurosci. 4, 1944 (1984).

    Article  CAS  Google Scholar 

  13. Kordeli, E., Lambert, S. & Bennett, V. AnkyrinG: a new ankyrin gene with neural-specific isoforms localized at the initial segment and Node of Ranvier. J. Biol. Chem. 270, 2352–2359 (1995).

    Article  CAS  Google Scholar 

  14. Butler, M. H. et al. Amphiphysin II (SH3P9;BIN1), a member of the amphiphysin/Rvs family, is concentrated in the cortical cytomatrix of axon initial segments and Nodes of Ranvier in brain and around T tubules in skeletal muscle. J. Cell Biol. 137, 1355–1367 (1997).

    Article  CAS  Google Scholar 

  15. Davis, J. Q. & Bennett, V. Ankyrin binding activity shared by neural fascin/L1/NrCAM family of nervous system cell adhesion molecules abolishes ankyrin binding and increases lateral mobility of neurofascin. J. Cell Biol. 137, 703–714 (1997).

    Article  Google Scholar 

  16. Garver, T. D., Ren, Q., Tuvia, S. & Bennett, V. Tyrosine phosphorylation at a site highly conserved in the L1 family of adhesion molecules abolishes ankyrin binding and increases lateral mobility of neurofascin. J. Cell Biol. 137, 703–714 (1997).

    Article  CAS  Google Scholar 

  17. Zhou, D. et al. Ankyrin G is required for clustering of voltage-gated Na channels at axon initial segments and for normal action potential firing. J. Cell Biol. 143, 1295 (1998).

    Article  CAS  Google Scholar 

  18. Angelides, K. J., Elmer, L. W., Loftus, D. & Elson, E. Distribution and lateral mobility of voltage-dependent sodium channels in neurons. J. Cell Biol. 106, 1911 (1988).

    Article  CAS  Google Scholar 

  19. Dargent, B. et al. Targeting of the voltage-dependent sodium channel to the axon of cultured hippocampal neurons. Soc. Neurosci. Abstr. 24, 1078 (1998).

    Google Scholar 

  20. Sheetz, M. P. Glycoprotein mobility and dynamic domains in fluid membranes. Annu. Rev. Biophys. Biomol. Struct. 22, 417 (1993).

    Article  CAS  Google Scholar 

  21. Bartles, J. R. The spermatid membrane comes of age. Trends Cell Biol. 5, 400–407 (1995).

    Article  CAS  Google Scholar 

  22. Banker, G. & Goslin, K. (eds) Culturing Nerve Cells(MIT Press, Cambridge, Massachusetts, (1997).

    Google Scholar 

  23. Brewer, G. J., Torricelli, J. R., Evege, E. K. & Price, P. J. Optimized survival of hippocampal neurons in B-27 supplemented Neurobasal, a new serum-free medium combination. J. Neurosci. Res. 35, 567–576 (1993).

    Article  CAS  Google Scholar 

  24. Thompson, C., Lin, C. H. & Forscher, P. An Aplysia cell adhesion molecule associated with site-directed actin filament assembly in neuronal growth cones. J. Cell Sci. 109, 2843–2854 (1996).

    CAS  PubMed  Google Scholar 

  25. Forscher, P. & Smith, S. J. Actions of cytochalasins on the organization of actin filaments and microtubules in a neuronal growth cone. J. Cell Biol. 107, 1505–1516 (1988).

    Article  CAS  Google Scholar 

  26. Jareb, M. & Banker, G. The polarized sorting of membrane proteins expressed in cultured hippocampal neurons using viral vectors. Neuron 20, 855 (1998).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank P. de Camilli, P. Sonderegger and V. Bennett for sharing their reagents; P.deCamilli, D. Suter and members of I.M.'s laboratory for adivce and stimulating discussions; and F.Solomon, S. Cash, M. Velleca, H. Fölsch and R. Kroschewski for critical reading of the manuscript.

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Correspondence to Ira Mellman.

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Winckler, B., Forscher, P. & Mellman, I. A diffusion barrier maintains distribution of membrane proteins in polarized neurons. Nature 397, 698–701 (1999). https://doi.org/10.1038/17806

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