Periodic actin structures in neuronal axons are required to maintain microtubules

Mol Biol Cell. 2017 Jan 15;28(2):296-308. doi: 10.1091/mbc.E16-10-0727. Epub 2016 Nov 23.

Abstract

Axons are cable-like neuronal processes wiring the nervous system. They contain parallel bundles of microtubules as structural backbones, surrounded by regularly spaced actin rings termed the periodic membrane skeleton (PMS). Despite being an evolutionarily conserved, ubiquitous, highly ordered feature of axons, the function of PMS is unknown. Here we studied PMS abundance, organization, and function, combining versatile Drosophila genetics with superresolution microscopy and various functional readouts. Analyses with 11 actin regulators and three actin-targeting drugs suggest that PMS contains short actin filaments that are depolymerization resistant and sensitive to spectrin, adducin, and nucleator deficiency, consistent with microscopy-derived models proposing PMS as specialized cortical actin. Upon actin removal, we observed gaps in microtubule bundles, reduced microtubule polymerization, and reduced axon numbers, suggesting a role of PMS in microtubule organization. These effects become strongly enhanced when carried out in neurons lacking the microtubule-stabilizing protein Short stop (Shot). Combining the aforementioned actin manipulations with Shot deficiency revealed a close correlation between PMS abundance and microtubule regulation, consistent with a model in which PMS-dependent microtubule polymerization contributes to their maintenance in axons. We discuss potential implications of this novel PMS function along axon shafts for axon maintenance and regeneration.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Actins / metabolism*
  • Actins / physiology
  • Animals
  • Axons / metabolism
  • Axons / physiology*
  • Cells, Cultured
  • Cytoskeleton / metabolism
  • Drosophila / metabolism
  • Drosophila Proteins / metabolism
  • Microfilament Proteins / metabolism
  • Microtubule-Associated Proteins / metabolism
  • Microtubules / metabolism
  • Microtubules / physiology*
  • Neurons / metabolism
  • Tubulin / metabolism

Substances

  • Actins
  • Drosophila Proteins
  • Microfilament Proteins
  • Microtubule-Associated Proteins
  • Tubulin