Phospholipase A activities in embryonic palate mesenchyme cells in vitro

Incubation of palate mesenchyme cells in vitro in the presence of [3H]arachidonate and [14C]palmitate resulted in incorporation of radiolabel into all major families of phospholipids. Almost all (more than 90%) of the 3H and about one-half of the 14C in the phospholipids were in the sn-2 position. The [14C] fatty acid in the sn-2 position was saturate. When [14C]stearate was added to the culture medium with [3H]arachidonate, 30-40% of the total 14C in phospholipids was in the sn-2 position. 80% of the 14C in the sn-2 position was found in unsaturated fatty acids. Hydrolysis of phospholipids could be demonstrated at acid, neutral and alkaline pH. Calcium stimulated phospholipase activity at neutral and alkaline pH, but inhibited hydrolytic activity at acid pH. Radiolabeled lysophospholipid indicative of phospholipase A2 activities accumulated at acid pH, whereas little, if any, radiolabeled lysophospholipids accumulated at neutral and alkaline pH. Quantitative analysis revealed the production of some lysophosphatidylethanolamine at alkaline pH.