Intracellular mechanisms involved in short-term regulation of net protein synthesis in pancreatic acini.
The mechanisms regulating the net synthesis of digestive enzymes during short-term stimulation by agonists were examined in pancreatic acini isolated from the rat. Dispersed pancreatic acini were stimulated for up to 60 min with various concentrations of cholecystokinin octapeptide (CCK-OP), carbachol, A23187, 4 beta-phorbol 12-myristate 13-acetate (PMA). The effects of these agonists on net protein synthesis was determined by measuring the incorporation of [3H]leucine or [35S]methionine into protein. Carbachol, PMA, A23187 and concentrations of CCK-OP of 100 pM and greater caused inhibition of protein synthesis. Fluorography of [35S]methionine labeled acinar cell proteins separated by one-dimensional SDS-polyacrylamide gel electrophoresis demonstrated that the agonists inhibited the synthesis of the digestive enzymes. Northern blot analysis using cDNA probes revealed that CCK-OP, carbachol and PMA did not alter the cellular content of amylase, lipase and elastase mRNA. The protein kinase C inhibitors 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) and staurosporine failed to reverse the inhibitory effects of CCK-OP, carbachol and PMA on protein synthesis. CCK-OP and PMA activated phospholipase A (PLA) which liberated lysophosphatidylcholine (LPC) and free fatty acids from membrane phosphatidylcholine. Exogenously added PLA2 (Naja naja venom) inhibited protein synthesis and increased LPC to a similar extent as CCK and PMA. The results suggest that the inhibitory effects of CCK and carbachol on net protein synthesis are due to their effects on intracellular calcium and PLA-mediated breakdown of phosphatidylcholine rather than protein kinase C activation.
Perkins
P S
PS
Department of Medicine, Veterans Administration Medical Center, San Diego, CA 92161.
Bahrami
L H
LH
Lenhard
L W
LW
Pandol
S J
SJ
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000470:Alkaloids / Q000494:pharmacology; D000681:Amylases / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D015342:DNA Probes; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D004798:Enzymes / Q000096:biosynthesis*; D007546:Isoquinolines / Q000494:pharmacology; D008049:Lipase / Q000096:biosynthesis / Q000235:genetics; D009693:Nucleic Acid Hybridization; D010179:Pancreas / Q000187:drug effects / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000096:biosynthesis / Q000235:genetics; D010713:Phosphatidylcholines / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism / Q000494:pharmacology; D054467:Phospholipases A2; D010879:Piperazines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D051381:Rats; D011919:Rats, Inbred Strains; D012844:Sincalide / Q000494:pharmacology; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000470:Alkaloids; D015342:DNA Probes; D004798:Enzymes; D007546:Isoquinolines; D010713:Phosphatidylcholines; D010879:Piperazines; D000001:Calcimycin; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D002217:Carbachol; D011493:Protein Kinase C; D008049:Lipase; D010741:Phospholipases A; D054467:Phospholipases A2; D000681:Amylases; D010196:Pancreatic Elastase; D019311:Staurosporine; D012844:Sincalide; D013755:Tetradecanoylphorbol Acetate