Association of Myosin Va and Schwann cells-derived RNA in mammal myelinated axons, analyzed by immunocytochemistry and confocal FRET microscopy

Methods. 2014 Mar 15;66(2):153-61. doi: 10.1016/j.ymeth.2013.06.007. Epub 2013 Jun 19.

Abstract

Evidence from multiple sources supports the hypothesis that Schwann cells in the peripheral nervous system transfer messenger RNA and ribosomes to the axons they ensheath. Several technical and methodological difficulties exist for investigators to unravel this process in myelinated axons - a complex two-cell unit. We present an experimental design to demonstrate that newly synthesized RNA is transferred from Schwann cells to axons in association with Myosin Va. The use of quantitative confocal FRET microscopy to track newly-synthesized RNA and determine the molecular association with Myosin Va, is described in detail.

Keywords: Axon; Myosin Va; Quantitative FRET microscopy; RNA cell-to-cell transfer; RNA transport; Schwann cell.

MeSH terms

  • Animals
  • Axons / metabolism*
  • Fluorescence Resonance Energy Transfer
  • Immunohistochemistry
  • Microscopy, Confocal
  • Myosin Heavy Chains / metabolism*
  • Myosin Type V / metabolism*
  • Peripheral Nerves / cytology
  • Peripheral Nerves / metabolism
  • RNA Transport
  • RNA, Messenger / metabolism*
  • Ranvier's Nodes / metabolism*
  • Rats
  • Schwann Cells / metabolism

Substances

  • Myo5a protein, rat
  • RNA, Messenger
  • Myosin Type V
  • Myosin Heavy Chains