Ca2+ release triggered by inositol trisphosphate (Ins(1,4,5)P3) has been measured in saponin-permeabilized hepatocytes with 45Ca2+ or Quin 2. The initial rate of Ca2+ release was not greatly affected by the incubation temperature (175 +/- 40 pmol X s-1 X mg dry weight-1, at 30 degrees C versus 133 +/- 24 pmol X s-1 X mg dry weight-1 at 4 degrees C). The amount of Ca2+ released by Ins(1,4,5)P3 was not affected by pH (6.5-8.0). La3+ (100 microM) markedly inhibited the effect of 1 microM Ins(1,4,5)P3. The possibility that La3+ chelates Ins(1,4,5)P3 cannot be excluded since the effect of La3+ could be overcome by increasing the Ins(1,4,5)P3 concentration. Ins(1,4,5)P3-mediated Ca2+ release showed a requirement for permeant cations in the incubation medium. Optimal release was observed with potassium gluconate. Other monovalent cations, with the exception of Li+, can substitute for K+. Permeant anions, at concentrations above 40 mM, inhibited Ca2+ release produced by Ins(1,4,5)P3. Cl-, Br-, I-, and SO2-4 were equally effective as inhibitors. Ins(1,4,5)P3 also caused the release of 54Mn2+ and 85Sr2+ accumulated by the permeabilized hepatocytes. Our results are consistent with Ins(1,4,5)P3 promoting the membrane translocation of divalent cations through an ion channel rather than an ion carrier. The translocation of positive charge through this channel is balanced by ancillary movements of monovalent cations and anions across the reticular membranes. The transport systems responsible for these compensatory ion movements may represent a potential site for the regulation of the hormone-mediated Ca2+ signal.