Receptor-operated Ca2+ influx and its association with the Src family in secretagogue-stimulated pancreatic acini.
We investigated signal transduction between receptor-operated Ca(2+) influx (ROCI) and Src-related nonreceptor protein tyrosine kinase (PTK) in rat pancreatic acini. CCK and the Ca(2+) ionophore enhanced the Src-related PTK activity, whereas the high-affinity CCK-A receptor agonists, fibroblast growth factor (FGF), and the protein kinase C (PKC) activator had no or little effect. This increase was abolished by eliminating [Ca(2+)](o), loading of the intracellular Ca(2+) chelator, and administering the PTK inhibitor genistein. While genistein inhibited extracellular Ca(2+) or Mn(2+) entry induced by CCK and carbachol, it did not affect intracellular Ca(2+) release and oscillations. CCK dose-dependently increased the Src phosphotransferase activity, which was abolished by inhibitors of G(q) protein, phospholipase C (PLC), and Src, but not by the calmodulin kinase (CaMK) inhibitor. Intensities of the Src band and amounts of tyrosine phosphorylated Src were enhanced by CCK stimulation. Thus, Src cascades appear to be coupled to the low-affinity CCK-A receptor and utilize G(q)-PLC pathways for their activation, independent of PKC and CaMK cascades. The low-affinity CCK-A receptor regulates ROCI via mediation of Src-related PTK and activates Src pathways to cause [Ca(2+)](o)-dependent pancreatic exocytosis.
Tsunoda
Yasuhiro
Y
Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA. ytsunoda@umich.edu
Yoshida
Hitoshi
H
Nozu
Fumihiko
F
D000681:Amylases / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000737:chemistry / Q000378:metabolism*; D002417:Cattle; D002766:Cholecystokinin / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005346:Fibroblast Growth Factors / Q000235:genetics / Q000494:pharmacology; D016391:Genes, src / Q000187:drug effects; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D008297:Male; D010179:Pancreas / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D015240:Phorbol 12,13-Dibutyrate / Q000031:analogs & derivatives / Q000494:pharmacology; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D043886:Receptor, Cholecystokinin A / Q000819:agonists; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000502:physiology*; D054440:Receptors, Ghrelin; D011994:Recombinant Proteins / Q000494:pharmacology; D012969:Sodium Fluoride / Q000494:pharmacology; D014443:Tyrosine / Q000378:metabolism
D004791:Enzyme Inhibitors; D043886:Receptor, Cholecystokinin A; D043562:Receptors, G-Protein-Coupled; D054440:Receptors, Ghrelin; D011994:Recombinant Proteins; D015240:Phorbol 12,13-Dibutyrate; D014443:Tyrosine; D015759:Ionomycin; D005346:Fibroblast Growth Factors; D012969:Sodium Fluoride; D002766:Cholecystokinin; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C; D000681:Amylases; D002118:Calcium