Inhibition of inositol 1,4,5-trisphosphate-mediated Ca2+ release by Ca2+ in cells from peripheral tissues.

Abstract

Permeabilized cells attached to culture plates were used to evaluate the inhibition of inositol 1,4,5-trisphosphate-mediated release (IPMCR) by Ca2+. In AR42J cells, a pancreatic acinar cell line, when permeabilization and Ca2+ uptake were carried out at low ionized Ca2+ (0.06 microM), Ca2+ had little effect on IPMCR. On the other hand, when permeabilization and Ca2+ uptake were performed at 5 microM Ca2+, IPMCR was inhibited by Ca2+ with an apparent affinity of 0.24 microM. This inhibition could be modified by exposing the cytosol of permeabilized cells to low Ca2+. Hence, permeabilizing the cells in the presence of 5 microM Ca2+ and then exposing them to Ca2+ concentrations between 0.01 and 5 microM before washing and Ca2+ uptake in the presence of 5 microM Ca2+ resulted in a Ca2(+)-dependent loss of inhibitory activity. The loss of inhibitory activity occurred with an apparent affinity for Ca2+ of 0.21 microM. A similar phenomenon with a comparable apparent dissociation constant for Ca2+ was found with three other cell types from peripheral tissues: the osteosarcoma cell line UMR-106-01, the kidney inner medullary cell line IMCD, and primary culture of urinary bladder smooth muscle cells. The properties of inhibition of IPMCR by Ca2+ in cells from peripheral tissues differ from those previously described in neuronal tissues and suggest that a different factor(s) mediates the inhibition of IPMCR by Ca2+ in cells from peripheral and neuronal tissues.