Agonist-activated, ryanodine-sensitive, IP3-insensitive Ca2+ release channels in longitudinal muscle of intestine

We have previously shown that Ca2+ mobilization in longitudinal muscle is not mediated by inositol 1,4,5-trisphosphate (IP3) and depends on an obligatory influx of Ca2+. The present study examined whether Ca2+ influx activates ryanodine-sensitive Ca2+ channels to cause Ca(2+)-induced Ca2+ release. Ryanodine bound with high affinity to longitudinal muscle cells [dissociation constant (Kd) 7.3 +/- 0.3 nM] and microsomes (Kd 7.5 +/- 0.4 nM) and induced concentration-dependent 45Ca2+ efflux [50% effective concentration (EC50) 1.3 +/- 0.5 nM], increase in cytosolic free Ca2+ (EC50 2.0 +/- 0.7 nM), and contraction (EC50 0.9 +/- 0.2 nM) but had no effect in circular muscle cells. Ryanodine binding and ryanodine-induced Ca2+ release were enhanced by caffeine and inhibited by dantrolene and ruthenium red but were not affected by IP3 or heparin. Changes in Ca2+ concentration (50-500 nM) caused Ca2+ release from permeabilized longitudinal but not circular muscle cells loaded with 45Ca2+. The contractile agonist cholecystokinin-8 elicited 45Ca2+ efflux in both circular and longitudinal muscle cells; efflux in longitudinal muscle cells was abolished by Ca2+ channel blockers and by pretreatment of the cells with ryanodine. Pretreatment with thapsigargin abolished agonist-induced 45Ca2+ efflux in both cell types. We conclude that ryanodine-sensitive IP3-insensitive Ca2+ release channels with properties similar to those in cardiac muscle are present in longitudinal but not circular muscle cells of intestine and that agonist-mediated Ca2+ influx activates these channels, leading to Ca(2+)-induced Ca2+ release.