Pepsinogen secretion from streptolysin O-permeabilized chief cells from guinea pig stomach.
To evaluate directly the actions of cellular mediators on pepsinogen secretion, a nearly homogeneous population of dispersed chief cells was permeabilized using the bacterial toxin streptolysin O (30 IU/ml for 10 min). This resulted in the release of > 60% of cellular lactate dehydrogenase activity, whereas cellular pepsinogen levels were not altered. Examination of permeabilized cells using light and electron microscopy revealed preservation of cellular polarity and organelles. Pepsinogen secretion from permeabilized chief cells could be stimulated with increasing concentrations of calcium (300 nM to 10 microM), adenosine 3',5'-cyclic monophosphate (cAMP; 1 microM to 1 mM), phorbol 12-myristate 13-acetate (10 nM to 1 microM), or by addition of carbamylcholine (0.1 mM) to the incubation solution. In the absence of calcium [0.4 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid], activators of protein kinase C stimulated a two- to threefold increase in pepsinogen secretion, and potentiation of secretion was observed when these agents were combined with 0.3 and 1.0 microM calcium. In contrast to the effects of activators of protein kinase C, cAMP-induced pepsinogen secretion was observed only with calcium concentrations > or = 100 nM. Combinations of increasing concentrations of cAMP and calcium resulted in an additive secretory response. This study indicates the utility of streptolysin O-permeabilized chief cells for the study of signal transduction mechanisms that mediate pepsinogen secretion.
Raffaniello
R D
RD
Department of Medicine, State University of New York Health Science Center, Brooklyn 11203-2098.
Raufman
J P
JP
D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D001426:Bacterial Proteins; D002118:Calcium / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D002463:Cell Membrane Permeability / Q000187:drug effects; D000242:Cyclic AMP / Q000494:pharmacology; D005753:Gastric Mucosa / Q000166:cytology / Q000378:metabolism*; D006168:Guinea Pigs; D008297:Male; D010435:Pepsinogens / Q000378:metabolism*; D013301:Streptolysins / Q000494:pharmacology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D001426:Bacterial Proteins; D010435:Pepsinogens; D013301:Streptolysins; C025964:streptolysin O; D000255:Adenosine Triphosphate; D002217:Carbachol; D000242:Cyclic AMP; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium