Phorbol ester stimulation of active anion secretion in intestine

Phorbol 12,13-dibutyrate (PDB) stimulates electrogenic anion secretion in rabbit and chicken distal ileum. The 50% effective dose in each case was 15 nM, and the maximal short-circuit current (Isc) responses seen at 10(-6) M were 20 and 140 microA/cm2, respectively. Isc was also stimulated by the diacylglycerol 1-oleoyl-2-acetyl glycerol (10(-5) M) but not by the inactive phorbol ester analogue 4 alpha-phorbol 12,13-didecanoate. Removal of Ca from the serosal bathing medium and pretreatment of tissues with atropine (10(-6) M) or tetrodotoxin (2 X 10(-7) M) did not affect PDB-stimulated responses. PDB also did not alter basal intracellular free Ca levels in isolated chicken enterocytes loaded with quin 2 or basal adenosine 3',5'-cyclic monophosphate levels in intact tissues from rabbit and chicken. In the presence of PDB, Ca- and phospholipid-dependent phosphorylation of several proteins in particulate and soluble fractions from isolated chicken enterocytes were seen. Pretreatment of chicken and rabbit tissues with indomethacin or mepacrine inhibited the Isc responses to PDB and carbamylcholine, suggesting a role for prostaglandins. These results suggest that electrogenic anion secretion stimulated by phorbol esters and diacylglycerol may be mediated by activation of protein kinase C.