Low Concentrations of Arsenite Target the Intraluminal Inositol 1, 4, 5-Trisphosphate Receptor/Ryanodine Receptor Crosstalk to Significantly Elevate Intracellular Ca²

Arsenite is an established human carcinogen that induces cytotoxic and genotoxic effects through poorly defined mechanisms involving the formation of reactive oxygen species (ROS) and deregulated Ca²⁺ homeostasis. We used variants of the U937 cell line to address the central issue of the mechanism whereby arsenite affects Ca²⁺ homeostasis. We found that 6-hour exposure to the metalloid (2.5 μM), although not associated with an immediate or delayed toxicity, causes a significant increase in the intracellular Ca²⁺ concentration ([Ca²⁺]_i) through a mechanism characterized by the following components: 1) it was not affected by ROS produced under the same conditions; 2) a small amount of Ca²⁺ was mobilized from the inositol-1,4,5-trisphosphate receptor (IP₃R), and this response was not augmented by greater concentrations of the metalloid; 3) large amounts of Ca²⁺ were instead dose dependently mobilized from the ryanodine receptor (RyR) in response to IP₃R stimulation; 4) the cells maintained an intact responsiveness to agonist-stimulated Ca²⁺ mobilization from both channels; 5) arsenite, even at 5-10 µM, failed to directly mobilize Ca²⁺ from the RyR; and 6) arsenite failed to enhance Ca²⁺ release from the RyR under conditions in which the [Ca²⁺]_i was increased by either RyR agonists or ionophore-stimulated Ca²⁺ uptake. We therefore conclude that arsenite elevates the [Ca²⁺]_i by directly targeting the IP₃R and its intraluminal crosstalk with the RyR. This mechanism likely mediates mitochondrial superoxide formation, downstream damage on various biomolecules (including genomic DNA), and mitochondrial dysfunction/apoptosis eventually occurring after longer incubation to, or exposure to greater concentrations of, arsenite.