Streptolysin O-permeable pancreatic acini, which retain intact signaling systems, were used to study the regulation of the inositol 1,4,5- trisphosphate (Ins-1,4,5-P3)-activated Ca2+ channel during agonist stimulation, and antagonist inhibition. Stimulation of permeable cells with carbachol induced rapid Ca2+ release from internal stores. Addition of heparin prior to or after agonist stimulation inhibited the release, indicating the activation of the Ins-1,4,5-P3-dependent Ca2+ channels by the agonist. Termination of cell stimulation with the specific antagonist atropine rapidly inactivated the release channels. Channel inactivation by the antagonist was independent of Ins-1,4,5-P3 levels since (a) addition of atropine to carbachol-stimulated cells resulted in a slow hydrolysis of Ins- 1,4,5-P3, (b) addition of 10-fold excess Ins-1,4,5-P3 together with the agonist did not prevent channel inactivation by the antagonist, and (c) the antagonist inactivated Ca2+ release in the presence of saturating concentration of the nonhydrolyzable Ins-1,4,5-P3. Hence, the antagonist appears to stabilize the Ins-1,4,5-P3-activated Ca2+ channel in a state refractory to Ins-1,4,5-P3. These findings are the first direct evidence that the channel can exist in such a refractory state.
|Original language||English (US)|
|Number of pages||4|
|Journal||Journal of Biological Chemistry|
|State||Published - 1994|
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology