Protein kinase C potentiates cAMP-stimulated mouse duodenal mucosal bicarbonate secretion in vitro.
PKC has been shown to regulate epithelial Cl(-) secretion in a variety of models. However, the role of PKC in duodenal mucosal bicarbonate secretion is less clear. We aimed to investigate the role of PKC in regulation of duodenal mucosal bicarbonate secretion. Bicarbonate secretion by murine duodenal mucosa was examined in vitro in Ussing chambers using a pH-stat technique. PKC isoform expression and activity were assessed by Western blotting and in vitro kinase assays, respectively. PMA (an activator of PKC) alone had no effect on duodenal bicarbonate secretion or short-circuit current (I(sc)). When PMA and dibutyryl-cAMP (db-cAMP) were added simultaneously, PMA failed to alter db-cAMP-stimulated duodenal bicarbonate secretion or I(sc) (P > 0.05). However, a 1-h preincubation with PMA potentiated db-cAMP-stimulated duodenal bicarbonate secretion and I(sc) in a concentration-dependent manner (from 10(-8) to 10(-5)M) (P < 0.05). PMA preincubation had no effects on carbachol- or heat-stable toxin-stimulated bicarbonate secretion. Western blot analysis revealed that PKCalpha, -gamma, -epsilon, -, -micro, and -iota/lambda were expressed in murine duodenal mucosa. Ro 31-8220 (an inhibitor active against PKCepsilon, -alpha, -beta, and -gamma), but not Gö 6983 (an inhibitor active against PKCalpha, -gamma, -beta, and -delta), reversed the potentiating effect of PMA on db-cAMP-stimulated bicarbonate secretion. PMA also time- and concentration-dependently increased the activity of PKCepsilon, an effect that was prevented by Ro 31-8220 but not Gö 6983. These results demonstrate that activation of PKC potentiates cAMP-stimulated duodenal bicarbonate secretion, whereas it does not modify basal secretion. The effect of PKC on cAMP-stimulated bicarbonate secretion is mediated by the PKCepsilon isoform.
Tuo
Bi-Guang
BG
Univ. of California, San Diego Medical Center, Div. of Gastroenterology, 8414, 200 W. Arbor Dr., San Diego, CA 92103-8413, USA.
Chow
Jimmy Y C
JY
Barrett
Kim E
KE
Isenberg
Jon I
JI
D000818:Animals; D001639:Bicarbonates / Q000378:metabolism*; D003994:Bucladesine / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D004386:Duodenum / Q000187:drug effects / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism*; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism; D051379:Mice; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051744:Protein Kinase C-epsilon; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D001639:Bicarbonates; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D003994:Bucladesine; D000242:Cyclic AMP; C497256:Prkce protein, mouse; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon; D013755:Tetradecanoylphorbol Acetate