Role of calcium in monitor peptide-stimulated cholecystokinin release from perifused intestinal cells.
Monitor peptide stimulates cholecystokinin (CCK) release from the intestine, but the cellular mechanisms responsible for this effect are uncertain. In the present study, the roles of membrane potential difference and calcium influx in monitor peptide-mediated CCK release were examined in a perifusion system containing isolated mucosal cells from the rat duodenum. This method represents an in vitro system in which CCK-releasing cells can be challenged with secretagogues or other maneuvers to study the dynamics of hormone secretion. High concentrations of KCl (50 mM), which reduce electrical potential difference across the cell membrane, caused the release of CCK. This effect was inhibited by the calcium channel blocker MnCl2. Monitor peptide stimulated CCK release in a dose-dependent manner at concentrations from 3 x 10(-12) to 3 x 10(-8) M. The requirement for extracellular calcium in secretagogue-stimulated release of CCK was investigated using ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), a calcium chelator, and MnCl2. A calcium-free environment supplemented with 2 mM EGTA completely inhibited CCK secretion in response to stimulatory doses of monitor peptide. CCK secretion was restored when calcium was reintroduced into the system. Similarly, MnCl2 completely blocked monitor peptide-stimulated CCK release. These data indicate that membrane depolarization and monitor peptide stimulate the release of CCK through calcium-dependent mechanisms, suggesting that increases in intracellular calcium within CCK cells are likely to be important in CCK release.
Bouras
E P
EP
Department of Medicine, Duke University Medical Center, Durham, North Carolina.
Misukonis
M A
MA
Liddle
R A
RA
D000818:Animals; D002118:Calcium / Q000502:physiology*; D002712:Chlorides; D002766:Cholecystokinin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004594:Electrophysiology; D005768:Gastrointestinal Hormones / Q000494:pharmacology; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D008345:Manganese / Q000494:pharmacology; D017895:Manganese Compounds; D010477:Perfusion; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000494:pharmacology*
D002712:Chlorides; D005768:Gastrointestinal Hormones; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D017895:Manganese Compounds; C504705:Spink1 protein, rat; D008345:Manganese; D014359:Trypsin Inhibitor, Kazal Pancreatic; D002766:Cholecystokinin; C025340:manganese chloride; D002118:Calcium