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It is well known that mechanical stimulation elicits Ca responses in various cell types. Touch stimulation on a single cell induces an increase in the cytosolic Ca concentration ([Ca]i), which subsequently propagates to neighboring cells as an intercellular Ca wave. In addition to Ca entry through mechanosensitive channels, this Ca response is thought to be induced, at least in part, by inositol 1,4,5-trisphosphate (IP3)-dependent Ca release from intracellular Ca stores (1-3). However, this hypothesis has not been tested directly due to a lack of the quantitative method to measure the cytosolic concentration of IP3 ([IP3]i) in a single living cell. Recently, we developed a FRET-based IP3 biosensor, LIBRAvIII, and a method to measure [IP3]i quantitatively in the single living cell (4). In order to examine the involvement of IP3 in mechanical stimulation-induced Ca responses, we monitored IP3 and Ca responses simultaneously in HSY-EA1 cells, a human parotid cell line, using LIBRAvIII and the Ca indicator fura-2.

Monitoring of IP3 dynamics during the mechanical stimulation-induced intra- and intercellular Ca2+ waves in HSY human parotid cell line