The Nogo-66 receptor NgR1 is required only for the acute growth cone-collapsing but not the chronic growth-inhibitory actions of myelin inhibitors

J Neurosci. 2007 Jul 4;27(27):7117-24. doi: 10.1523/JNEUROSCI.1541-07.2007.

Abstract

Neuronal Nogo-66 receptor 1 (NgR1) has been proposed to function as an obligatory coreceptor for the myelin-derived ligands Nogo-A, oligodendrocyte myelin glycoprotein (OMgp), and myelin-associated glycoprotein (MAG) to mediate neurite outgrowth inhibition by these ligands. To examine the contribution of neuronal NgR1 to outgrowth inhibition, we used two different strategies, genetic ablation of NgR1 through the germline and transient short hairpin RNA interference (shRNAi)-mediated knock-down. To monitor growth inhibition, two different paradigms were used, chronic presentation of substrate-bound inhibitor to measure neurite extension and acute application of soluble inhibitor to assay growth cone collapse. We find that regardless of the NgR1 genotype, membrane-bound MAG strongly inhibits neurite outgrowth of primary cerebellar, sensory, and cortical neurons. Similarly, substrate-bound OMgp strongly inhibits neurite outgrowth of NgR1 wild-type and mutant sensory neurons. Consistent with these results, shRNAi-mediated knock-down of neuronal NgR1 does not result in a substantial release of L-MAG (large MAG) inhibition. When applied acutely, however, MAG-Fc and OMgp-Fc induce a modest degree of growth cone collapse that is significantly attenuated in NgR1-null neurons compared with wild-type controls. Based on our findings and previous studies with Nogo-66, we propose that neuronal NgR1 has a circumscribed role in regulating cytoskeletal dynamics after acute exposure to soluble MAG, OMgp, or Nogo-66, but is not required for these ligands to mediate their growth-inhibitory properties in chronic outgrowth experiments. Our results thus provide unexpected evidence that the growth cone-collapsing activities and substrate growth-inhibitory activities of inhibitory ligands can be dissociated. We also conclude that chronic axon growth inhibition by myelin is mediated by NgR1-independent mechanisms.

Publication types

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

MeSH terms

  • Animals
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • GPI-Linked Proteins
  • Growth Cones / drug effects
  • Growth Cones / physiology*
  • Growth Inhibitors / pharmacology*
  • Growth Inhibitors / physiology
  • Mice
  • Mice, Mutant Strains
  • Myelin Proteins / antagonists & inhibitors
  • Myelin Proteins / physiology
  • Myelin Sheath / drug effects
  • Myelin Sheath / physiology*
  • Myelin-Associated Glycoprotein / physiology
  • Nogo Receptor 1
  • Nogo Receptor 2
  • PC12 Cells
  • Rats
  • Receptors, Cell Surface / antagonists & inhibitors
  • Receptors, Cell Surface / physiology*

Substances

  • GPI-Linked Proteins
  • Growth Inhibitors
  • Myelin Proteins
  • Myelin-Associated Glycoprotein
  • Nogo Receptor 1
  • Nogo Receptor 2
  • RTN4RL2 protein, human
  • Receptors, Cell Surface
  • Rtn4r protein, mouse