Cholecystokinin inhibits phosphatidylcholine synthesis via a Ca(2+)-calmodulin-dependent pathway in isolated rat pancreatic acini. A possible mechanism for diacylglycerol accumulation

• Published in: The Journal of biological chemistry 1991-11, Vol.266 (33), p.22246-22253
• Main Authors: Matozaki, T, Sakamoto, C, Nishisaki, H, Suzuki, T, Wada, K, Matsuda, K, Nakano, O, Konda, Y, Nagao, M, Kasuga, M
• Format: Article
• Language: English
• Subjects: Alkaloids - pharmacology; Animals; Bombesin - pharmacology; Bucladesine - pharmacology; Calcimycin - pharmacology; Calcium - physiology; Calmodulin - antagonists & inhibitors; Calmodulin - physiology; Carbachol - pharmacology; Choline - metabolism; Choline Kinase - metabolism; Choline-Phosphate Cytidylyltransferase; Diglycerides - metabolism; In Vitro Techniques; Kinetics; Male; Nucleotidyltransferases - metabolism; Pancreas - drug effects; Pancreas - metabolism; Phosphatidylcholines - antagonists & inhibitors; Phosphatidylcholines - biosynthesis; Protein Kinase C - antagonists & inhibitors; Rats; Rats, Inbred Strains; Secretin - pharmacology; Sincalide - pharmacology; Staurosporine; Sulfonamides - pharmacology; Tetradecanoylphorbol Acetate - pharmacology; Tritium
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)54561-0

The effects of cholecystokinin (CCK) and other pancreatic secretagogues on phosphatidylcholine (PC) synthesis were studied in isolated rat pancreatic acini. When acini were incubated with [3H]choline in the presence of 1 nM CCK-octapeptide (CCK8) for 60 min, the incorporations of [3H]choline into both water-soluble choline metabolites and PC in acini were reduced by CCK8 to 74 and 41% of control, respectively. Pulse-chase study revealed that CCK8 reduced both the disappearance of phosphocholine and the synthesis of PC. Other Ca(2+)-mobilizing secretagogues such as carbamylcholine, bombesin, and Ca2+ ionophore A23187 also reduced PC synthesis to the same extent as did CCK8. When combined with 1 nM CCK8, A23187 or carbamylcholine did not further inhibit PC synthesis. Furthermore, W-7 or W-5, a calmodulin antagonist, reversed the inhibition by CCK8 of PC synthesis, suggesting that a Ca(2+)-calmodulin-dependent pathway may be involved in CCK-induced inhibition of PC synthesis in acini. By contrast, neither cAMP-dependent secretagogues such as secretin and dibutyryl cAMP nor a phorbol ester had any effect on PC synthesis in acini. Staurosporine or H-7, a protein kinase C inhibitor, did not affect the inhibition by CCK of PC synthesis. The analysis of enzyme activity involved in PC synthesis via CDP-choline pathway showed that CCK treatment of acini reduced CTP:phosphocholine cytidylyltransferase activity in both cytosolic and particulate fraction, a finding consistent with the delayed disappearance of phosphocholine induced by CCK in pulse-chase study. By contrast, CCK treatment of acini did not alter the activities of choline kinase and phosphocholine transferase in acini. The extent of inhibition by CCK of cytidylyltransferase activity became much larger when subcellular fractions of acini were prepared in the presence of phosphatase inhibitors. In addition, W-7 reversed the inhibitory effect of CCK treatment on cytidylyltransferase activity in acini. When acini were labeled with [3H]myristic acid and chased, CCK8 (1 nM) reduced the synthesis of [3H]myristic acid-labeled PC to 27% of control after a 60-min chase period. This inhibition of PC synthesis induced by CCK was accompanied by a delayed disappearance of [3H]diacylglycerol, the radioactivity of which was 225% of control at 60 min. These results indicate that CCK inhibits PC synthesis by inducing both the reduction of choline uptake into acini and the inhibition of CTP:phosphocholine cytidylyltransferas