The incorporation and mobilization of [3H]arachidonic acid in lipids of human neuroblastoma cell lines, SK-N-SHF and LA-N-5, was studied. Essentially similar results were obtained with these two cell lines. Except for phosphatidylinositol which displayed the highest specific activity, the incorporation patterns within phospholipid classes tended to reflect phospholipid composition initially. However at later stages, counts in the acid-stable phosphatidylcholine plateaued and/or decreased while those of plasmenylethaniolamine and acid-stable phosphatidylethanolamine increased steadily. When cells were pulse-labelled with [3H]arachidonic acid and chased with fresh medium, there was a movement of label from diacyl (acid-stable) phosphatidylcholine to plasmenylethanolamine and diacyl (acid-stable) phosphatidylethanolamine. Plasmenylcholine did not appear to be involved in the arachidonyl group transfer. Under these chase conditions there was extensive turnover in the 32P-labelled polar headgroup of phosphatidylinositol but not in that of the other phospholipids. In both incorporation and chase studies involving [3H]arachidonic acid, a movement of arachidonyl groups from triacylglycerol to phospholipid could be observed. The results indicated that the patterns of incorporation and redistribution of arachidonic acid in human neuroblastoma cells were effectively regulated to favor lipids such as phosphatidylinositol and the subclasses of phosphatidylethanolamine. Possible mechanisms involved in these enrichment processes are discussed.