When administered in the diet to third instar Drosophila melanogaster larvae, short chain primary alcohols reduce phosphatidylcholine (PC) levels. The ethanol-induced reductions in larval PC may be in part due to an increase in the activity of PC-specific phospholipase D (PC-specific PLD, EC 18.104.22.168). PC-specific PLD not only hydrolyzes PC, but it also apparently catalyzes the formation of phosphatidylethanol. PC-specific PLD activity was also stimulated by 200 mM ethanol, methanol, isopropanol, n-butanol, and n-propanol. In vitro studies indicated that Drosophila PC-specific PLD activities were enhanced by submicromolar concentrations of Ca2+ and by GTP-gamma S. In vivo studies utilizing [14C]lyso-palmitoyl phosphatidylcholine indicated that dietary ethanol promoted the flux of label into triacylglycerol, 1,2 diacylglycerol, and fatty acid ethyl esters, while the label in PC decreased.