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J. Biol. Chem., Vol. 259, Issue 21, 12952-12955, 11, 1984

The effect of inositol trisphosphate on Ca2+ fluxes in insulin- secreting tumor cells

SK Joseph, RJ Williams, BE Corkey, FM Matschinsky and JR Williamson

An early event associated with the stimulation of various secretory cells is the breakdown of phosphatidylinositol 4,5-bisphosphate and the mobilization of cellular calcium. Hydrolysis of this inositol lipid by a phosphodiesterase produces inositol trisphosphate (InsP3), a small water-soluble molecule which may serve a messenger function to release Ca2+ from internal stores. In order to assess the role of inositol lipid breakdown in the stimulation of insulin secretion we have examined the effect of InsP3 on Ca2+ fluxes in three different insulin- secreting tumor cells permeabilized by the addition of saponin. A rapid, transient release of Ca2+ from a non-mitochondrial pool occurred upon addition of InsP3 to all three cell types. Half-maximal Ca2+ release from the RIN-1046-38 and RIN-m5F cells was obtained in the concentration range 0.1-0.2 microM. However, the cells obtained from a transplantable tumor of the Syrian hamster were far more sensitive to InsP3 with half-maximal release being observed at 0.025 microM. A partially purified preparation of vesicles was isolated from this tumor which retained its responsiveness to InsP3. Half-maximal Ca2+ release from the vesicles was obtained at 0.2 microM InsP3. Our data are consistent with a role for InsP3 in mediating the increase in cytosolic free Ca2+ which occurs in response to a number of stimuli that promote the secretion of insulin.
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