The incorporation of various labeled precursors into alkenylacyl, alkylacyl and diacyl phospholipids in rabbit alveolar macrophages was studied. The incorporation rates of the individual precursors were shown to be quite different among the three subclasses of phospholipids. [3H]Glycerol, [14C]16:0, [14C]18:1, [14C]18:2 and [32P]-orthophosphate were preferentially incorporated into choline glycerophospholipids (CGP), especially into diacyl glycerophosphocholine (GPC), indicating that the de novo synthesis of diacyl GPC is extremely high. Considerable portions of the radioactivities of [14C]16:0, [14C]18:1, [14C]18:2 and [32P]orthophosphate were also found in alkylacyl GPC, the incorporation being higher than or comparable to that in the case of diacyl glycerophosphoethanolamine (GPE). We then examined the activities of cholinephosphotransferase and ethanol-aminephosphotransferase, and found that the activity of cholinephosphotransferase was remarkably high in macrophage microsomes compared with that in microsomes from several other tissues. This suggests that diradylglycerols were preferentially utilized by choline-phosphotransferase, which is consistent with the results obtained for intact cells. We confirmed that a considerably higher amount of diacyl GPC as well as alkylacyl GPC was formed through this enzyme reaction with macrophage microsomes than with brain microsomes. The high formation of alkylacyl GPC could be responsible, at least in part, for the accumulation of this unique ether phospholipid, a stored precursor form of platelet-activating factor in macrophages.