STIMULATION OF ANANDAMIDE BIOSYNTHESIS IN N-18TG2 NEUROBLASTOMA-CELLSBY DELTA(9)-TETRAHYDROCANNABINOL (THC)

A concentration-related stimulation of anandamide (arachidonylethanola mide) synthesis by Delta(9)-tetrahydrocannabinol (THC) was observed in N-18TG2 neuroblastoma cells. Anandamide was detected and measured usi ng an approach in which[H-3]arachidonic acid and [C-14]ethanolamine we re incorporated into the phospholipids of subconfluent monolayers of c ells and the radiolabeled products were analyzed by TLC following agon ist exposure. Both precursors showed similar concentration-response re lationships and time dependencies consistent with the production of a product containing both the ethanolamine and arachidonyl moieties. The radiolabeled product also migrated together with authentic anandamide on two-dimensional TLC, confirming its identity as arachidonylethanol amide. Approximately two-thirds of the observed synthesis could be inh ibited by 1 mu M wortmannin, an agent previously reported to inhibit T HC-stimulated arachidonic acid release. These findings are in agreemen t with reports showing that THC can mobilize phospholipid bound arachi donic acid, leading to the production of other eicosanoids.