Calcium-dependent enzyme activation and vacuole formation in the apical granular region of pancreatic acinar cells.
The pancreatic acinar cell produces powerful digestive enzymes packaged in zymogen granules in the apical pole. Ca(2+) signals elicited by acetylcholine or cholecystokinin (CCK) initiate enzyme secretion by exocytosis through the apical membrane. Intracellular enzyme activation is normally kept to a minimum, but in the often-fatal human disease acute pancreatitis, autodigestion occurs. How the enzymes become inappropriately activated is unknown. We monitored the cytosolic Ca(2+) concentration ([Ca(2+)](i)), intracellular trypsin activation, and its localization in isolated living cells with specific fluorescent probes and studied intracellular vacuole formation by electron microscopy as well as quantitative image analysis (light microscopy). A physiological CCK level (10 pM) eliciting regular Ca(2+) spiking did not evoke intracellular trypsin activation or vacuole formation. However, stimulation with 10 nM CCK, evoking a sustained rise in [Ca(2+)](i), induced pronounced trypsin activation and extensive vacuole formation, both localized in the apical pole. Both processes were abolished by preventing abnormal [Ca(2+)](i) elevation, either by preincubation with the specific Ca(2+) chelator 1, 2-bis(O-aminophenoxy)ethane-N,N-N',N'-tetraacetic acid (BAPTA) or by removal of external Ca(2+). CCK hyperstimulation evokes intracellular trypsin activation and vacuole formation in the apical granular pole. Both of these processes are mediated by an abnormal sustained rise in [Ca(2+)](i).
Raraty
M
M
Medical Research Council Secretory Control Research Group, Physiological Laboratory, and Departments of Surgery and Veterinary Preclinical Sciences, University of Liverpool, Liverpool L69 3BX, United Kingdom.
Ward
J
J
Erdemli
G
G
Vaillant
C
C
Neoptolemos
J P
JP
Sutton
R
R
Petersen
O H
OH
D000818:Animals; D002118:Calcium / Q000502:physiology*; D020013:Calcium Signaling / Q000502:physiology*; D002766:Cholecystokinin / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D003600:Cytosol / Q000378:metabolism; D004789:Enzyme Activation; D007425:Intracellular Membranes / Q000502:physiology; D008564:Membrane Potentials; D051379:Mice; D008928:Mitochondria / Q000502:physiology; D010179:Pancreas / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D014357:Trypsin / Q000378:metabolism*; D014617:Vacuoles / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure
D002766:Cholecystokinin; D014357:Trypsin; D002118:Calcium