Cholecystokinin octapeptide CCK-8 and carbachol reduce [(32)P]orthophosphate labeling of phosphatidylcholine without modifying phospholipase D activity in rat pancreatic acini.

We have studied phospholipase D activation in [(32)P]orthophosphoric acid-prelabeled rat pancreatic acini by measuring the formation of (32)P-phosphatidylalcohols as stimulated in the presence of ethanol or butanol. A small but significant and time-dependent basal accumulation of [(32)P]phosphatidylethanol and [(32)P]phosphatidylbutanol was detected, which was further stimulated by phorbol myristate acetate, orthovanadate and pervanadate. However, the secretagogues cholecystokinin octapeptide and carbachol did not enhance basal accumulation of (32)P-phosphatidylalcohol, yet they decreased [(32)P]phosphatidylcholine content and stimulated the generation of [(32)P]phosphatidic acid. Our results stress the need to examine the transphosphatidylation reaction as well as agonist effects on the synthesis of phosphatidylcholine in order to assess unambiguously phospholipase D activity.