N-methyl-D-aspartate effects on the growth, morphology and cytoskeleton of individual neurons in vitro

Brain Res Dev Brain Res. 1992 Jun 19;67(2):301-8. doi: 10.1016/0165-3806(92)90231-k.

Abstract

Short-term (up to 5 h post-plating) cerebellar granule cell cultures were prepared from the week-old rat and maintained in a micro-incubator during time lapse video microscopy to examine normal and N-methyl-D-aspartate (NMDA)-evoked neurite extension. In untreated cultures growth of neurites was stochastic but proceeded at an average rate of 12.0 +/- 1.4 microns/h. Growth cone morphology was variable. The classical filopodia and lamellipodia possessing tips were motile or non-motile, while those processes ending in a club shape were rarely seen to extend. Individual growth cones passed through several different morphologies during growth. New processes extended more rapidly (13.0 +/- 1.7 microns/h) than those already present (9.0 +/- 0.5 microns/h). Addition of the NMDA receptor antagonist, aminophosphonovalerate (APV), caused a marked retraction of pre-existing processes. Stimulation of the receptor with 50 microM NMDA caused a marked increase in growth rate compared to controls (15.0 microns/h and 1.7 microns/h, respectively). When the presence of actin-rich structures was examined using rhodamine-phalloidin labelling it was found that NMDA increased the proportion of neuronal processes that possessed a growth cone by 28%. Conversely, inhibition of NMDA receptor activity with APV reduced the formation of lamellipodia from neuronal cell bodies.

MeSH terms

  • 2-Amino-5-phosphonovalerate / pharmacology
  • Animals
  • Cell Division / drug effects
  • Cells, Cultured
  • Cytoplasmic Granules / drug effects
  • Cytoplasmic Granules / ultrastructure
  • Cytoskeleton / drug effects
  • Cytoskeleton / ultrastructure*
  • N-Methylaspartate / pharmacology*
  • Neurites / drug effects
  • Neurites / ultrastructure*
  • Neurons / cytology*
  • Neurons / drug effects
  • Neurons / ultrastructure
  • Rats
  • Time Factors

Substances

  • N-Methylaspartate
  • 2-Amino-5-phosphonovalerate