Alcohol redirects CCK-mediated apical exocytosis to the acinar basolateral membrane in alcoholic pancreatitis.
The molecular mechanism of clinical alcohol-induced pancreatitis remains vague. We had reported that experimental high-dose cholecystokinin (CCK)-induced pancreatitis is in part because of excessive aberrant basolateral exocytosis. High-dose CCK caused Munc18c on basolateral plasma membrane (BPM) to dissociate from syntaxin (Syn)-4, activating Syn-4 to complex with plasma membrane (PM)-SNAP-23 and granule-VAMP to mediate basolateral exocytosis. We now hypothesize that alcohol could render the acinar cell BPM conducive to exocytosis by a similar mechanism. Weakly stimulating postprandial doses of alcohol (20-50 mM) inhibited postprandial low-dose CCK-stimulated secretion by blocking physiologic apical exocytosis and redirecting exocytosis to less-efficient basal PM (visualized by FM1-43 fluorescence imaging) and lateral PM sites (electron microscopy). Alcohol or low-dose CCK had no effect on PM-Munc18c, but alcohol preincubation enabled low-dose CCK to displace Munc18c from BPM, leading to SNARE complex assembly in the BPM. Similarly, alcohol diet-fed rats did not exhibit morphologic defects in the pancreas nor affected PM-Munc18c behavior, but subsequent intraperitoneal injections of low-dose CCK analog cerulein caused Munc18c displacement from BPM and cytosolic degradation, which contributed to pancreatitis. We conclude that alcohol induces BPM-Munc18c to become receptive to postprandial CCK-induced displacement into the cytosol, a process which facilitates SNARE complex assembly that in turn activates restricted BPM sites to become available for aberrant exocytosis into the interstitial space, where zymogen activation would take place and cause pancreatitis.
Lam
Patrick P L
PP
Department of Medicine, University Health Network, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
Cosen Binker
Laura I
LI
Lugea
Aurelia
A
Pandol
Stephen J
SJ
Gaisano
Herbert Y
HY
D000681:Amylases / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002492:Central Nervous System Depressants / Q000494:pharmacology / Q000633:toxicity; D002108:Ceruletide / Q000494:pharmacology; D002766:Cholecystokinin / Q000031:analogs & derivatives / Q000494:pharmacology*; D003600:Cytosol / Q000378:metabolism; D000431:Ethanol / Q000494:pharmacology* / Q000633:toxicity; D005089:Exocytosis / Q000187:drug effects*; D005765:Gastrointestinal Agents / Q000494:pharmacology; D015151:Immunoblotting; D008297:Male; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008954:Models, Biological; D051938:Munc18 Proteins / Q000378:metabolism; D046790:Pancreas, Exocrine / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D019512:Pancreatitis, Alcoholic / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D010446:Peptide Fragments / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D050600:SNARE Proteins / Q000378:metabolism; D012844:Sincalide / Q000031:analogs & derivatives / Q000494:pharmacology
D002492:Central Nervous System Depressants; D005765:Gastrointestinal Agents; D051938:Munc18 Proteins; D010446:Peptide Fragments; D050600:SNARE Proteins; C067408:cholecystokinin (26-32), Tyr-Gly-Nle(28,31) phenethyl ester-; D000431:Ethanol; D002108:Ceruletide; D002766:Cholecystokinin; D000681:Amylases; D012844:Sincalide