Determination of the phospholipid precursor of anandamide and other N-acylethanolamine phospholipids before and after sodium azide-induced toxicity in cultured neocortical neurons

J Neurochem. 2000 Aug;75(2):861-71. doi: 10.1046/j.1471-4159.2000.0750861.x.


Phospholipase D-mediated hydrolysis of N-acylethanolamine phospholipids (NAPEs) releases anandamide and other N-acylethanolamines, resulting in different actions at cellular targets in the CNS. Recently, we have demonstrated that these N-acyl lipids accumulate in cultured neocortical neurons subjected to sodium azide-induced cell injury. We here extend the information on the NAPE response, reporting on the composition of N-acylspecies of NAPE, employing a new methodological approach of HPLC-coupled electrospray ionization mass spectrometry. Exposure to sodium azide (5 mM) increased the total amount of NAPE threefold over control levels; however, no alteration of the relative composition of NAPE species was detected. The anandamide precursor (20 : 4-NAPE) constituted only 0.1% of all NAPEs detected in the neurons. Total NAPE species in control cells amounted to 956-1,060 pmol/10(7) cells. Moreover, we detected the presence of an unknown NAPE species with molecular weight identical to 20 : 4-NAPE. This may suggest the presence of a putative stereoisomer of the anandamide precursor with at least one trans-configured double bond in the N-arachidonoyl moiety. These results show that with the present method, neuronal NAPE species can be identified and quantified with respect to N-acyl composition, including a trans-isomer of the anandamide precursor. The anandamide precursor is up-regulated to the same extent as other NAPEs upon neuronal injury.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arachidonic Acids / metabolism*
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Embryo, Mammalian
  • Endocannabinoids
  • Mice
  • Mice, Inbred Strains
  • Neocortex / cytology
  • Neocortex / drug effects
  • Neocortex / metabolism*
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / metabolism*
  • Phosphatidylethanolamines / metabolism*
  • Phospholipase D / metabolism
  • Phospholipids / metabolism*
  • Plasmalogens / metabolism
  • Polyunsaturated Alkamides
  • Sodium Azide / toxicity*
  • Spectrometry, Mass, Secondary Ion


  • Arachidonic Acids
  • Endocannabinoids
  • Phosphatidylethanolamines
  • Phospholipids
  • Plasmalogens
  • Polyunsaturated Alkamides
  • Sodium Azide
  • Phospholipase D
  • anandamide