STED nanoscopy reveals the ubiquity of subcortical cytoskeleton periodicity in living neurons

Cell Rep. 2015 Mar 3;10(8):1246-51. doi: 10.1016/j.celrep.2015.02.007. Epub 2015 Feb 26.

Abstract

In the axons of cultured hippocampal neurons, actin forms various structures, including bundles, patches (involved in the preservation of neuronal polarity), and a recently reported periodic ring-like structure. Nevertheless, the overlaying organization of actin in neurons and in the axon initial segment (AIS) is still unclear, due mainly to a lack of adequate imaging methods. By harnessing live-cell stimulated emission depletion (STED) nanoscopy and the fluorescent probe SiR-Actin, we show that the periodic subcortical actin structure is in fact present in both axons and dendrites. The periodic cytoskeleton organization is also found in the peripheral nervous system, specifically at the nodes of Ranvier. The actin patches in the AIS co-localize with pre-synaptic markers. Cytosolic actin organization strongly depends on the developmental stage and subcellular localization. Altogether, the results of this study reveal unique neuronal cytoskeletal features.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / chemistry
  • Actins / metabolism
  • Animals
  • Cells, Cultured
  • Cytoskeleton / chemistry*
  • Cytoskeleton / metabolism
  • Dendrites / chemistry
  • Dendrites / metabolism
  • Female
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Nanotechnology
  • Nerve Net / physiology
  • Neurons / cytology
  • Neurons / metabolism*
  • Ranvier's Nodes / metabolism
  • Rats
  • Rats, Wistar

Substances

  • Actins
  • Fluorescent Dyes