Determination of the phospholipid precursor of anandamide and other N- acylethanolamine phospholipids before and after sodium azide-induced toxicity in cultured neocortical neurons
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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-acyl species 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 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.
Originalsprog | Engelsk |
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Tidsskrift | Journal of Neurochemistry |
Vol/bind | 75 |
Udgave nummer | 2 |
Sider (fra-til) | 861-871 |
Antal sider | 11 |
ISSN | 0022-3042 |
DOI | |
Status | Udgivet - 1 jan. 2000 |
ID: 45562245