The cholecystokinin analogues JMV-180 and CCK-8 stimulate phospholipase C through the same binding site of CCKA receptor in rat pancreatic acini

Elisabet Sarri*, Belén Ramos, Ginés M. Salido, Enrique Claro

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

5 Citations (Scopus)

Abstract

1. This study was designed to address the controversy related to the involvement of phospholipase C in the signalling pathway linked to CCKA receptor stimulation by the cholecystokinin analogue JMV-180, a full agonist for amylase release, in rat pancreatic acini. 2. JMV-180 was shown to stimulate phospholipase C by measuring the incorporation of [32P]-orthophosphoric acid ([32P]-Pi) into phosphatidic acid (PtdOH) and phosphatidylinositol (PtdIns). Both responses elicited by JMV-180 were time and concentration dependent. Maximal effects elicited by JMV-180 were 39.08±0.72 and 8.02±0.40% for the labelling of [32P]-PtdIns and [32P]-PtdOH, respectively, as compared to the maximal effects of CCK-8, a full agonist of the CCKA receptor. 3. [32P]-Pi incorporation into PtdOH and PtdIns was sensitive to lithium, demonstrating that both responses are a consequence of phospholipase C activation. However, since lithium blocks the phosphoinositide cycle by an uncompetitive mechanism, its effect was only apparent at high concentrations of CCK-8 (< 10 pM), which elicited stimuli above 20 and 60% of the maximal [32P]-PtdOH and [32P]-PtdIns labelling, respectively. 4. JMV-180 inhibited the incorporation of [32P]-Pi into PtdOH and PtdIns as stimulated by CCK-8, down to its own maximal effect. The estimated IC50 values for the inhibition curves were not significantly different from those calculated assuming the same single binding site for both agonists. These results indicated that the well established role of JMV-180 as a partial agonist for CCKA receptor-linked signalling responses, also applies for the stimulation of phospholipase C. 5. The comparison of CCK-8 and JMV-180 dose-response curves of amylase release to those of PtdIns and PtdOH labelling with [32P]-Pi showed the existence of an amplification mechanism between phospholipase C and amylase release for both agonists. 6. In conclusion, we show that JMV-180, as well as CCK-8, stimulate phospholipase C upon interaction with the same binding site at the CCKA receptor in rat pancreatic acini.

Original languageEnglish
Pages (from-to)1227-1234
Number of pages8
JournalBritish Journal of Pharmacology
Volume133
Issue number8
DOIs
Publication statusPublished - 2001

Keywords

  • Amylase release
  • CCK-8
  • JMV-180
  • Lithium
  • Pancreatic acini
  • Phosphoinositide cycle
  • Phospholipase C
  • Rat

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