CCK-stimulated changes in pancreatic acinar morphology are mediated by rho.
OBJECTIVE
Treatment of isolated pancreatic acini with high concentrations of cholecystokinin (CCK) is known to induce rapid changes in the cellular morphology. The signalling pathways remain to be characterized.


METHODS
Pancreatic acini were permeabilized by digitonin and incubated with various agents. The acinar morphology was investigated by microscopy. The activation of p125 focal adhesion kinase was determined by Western blot analysis. Amylase was measured photometrically.


RESULTS
The functionality of the permeabilized acini was tested by measuring stimulated amylase release. 300 microM GTP gamma S was almost as efficient as CCK to stimulate amylase release, while 300 microM GDP beta S inhibited the CCK-stimulated amylase release. Stimulation of permeabilized acini with 0.1 microM CCK induced similar morphological changes as in unpermeabilized acini. Incubation of permeabilized acini with GTP gamma S mimicked the CCK-induced changes, whereas a preincubation with GDP beta S prevented the CCK effects on the acinar morphology. Inhibition of the small G protein rho, which activates p125 focal adhesion kinase, by Clostridium botulinum C3 transferase also prevented the CCK-stimulated morphological changes. Preincubation of intact acini with cell-permeable inhibitors of protein kinase C, MEK or p38MAPK, or with the intracellular calcium chelator BAPTA/AM was without significant effect on the CCK-stimulated changes.


CONCLUSIONS
The CCK-induced morphological changes seem to be mediated by G protein signalling via the small G protein rho and the associated activation of p125 focal adhesion kinase.
Kiehne
Karlheinz
K
First Department of Internal Medicine, Christian Albrechts University, Kiel, Germany. kkiehne@1med.uni-kiel.de
Herzig
Karl H
KH
Fölsch
Ulrich R
UR
D000818:Animals; D015153:Blotting, Western; D002766:Cholecystokinin / Q000494:pharmacology*; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D022001:Focal Adhesions; D010179:Pancreas / Q000166:cytology / Q000187:drug effects*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D020741:rho GTP-Binding Proteins / Q000502:physiology*
D002766:Cholecystokinin; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496052:Ptk2 protein, rat; D020741:rho GTP-Binding Proteins