Spontaneous calcium responses of SF2 rat dental epithelial cells stably expressing the calcium sensor G-GECO

抄録

Genetically-encoded calcium indicators such as G-GECO are useful for studying Ca²⁺ responses during long-term processes. In this study, we employed a lentiviral vector and established a rat dental epithelial cell line that stably expressed G-GECO (SF2-G-GECO). Ca²⁺ imaging analysis under cell culture conditions revealed that SF2-G-GECO cells exhibited spontaneous Ca²⁺ responses, which could be classified into the following three major patterns depending on the cell density: localized Ca²⁺ responses at cell protrusions at a low density, a cell-wide spread of Ca²⁺ responses at a medium density, and Ca²⁺ responses in clusters of 3–20 cells at a high density. The P2Y receptor inhibitor suramin (10 μM), the ATP-degrading enzyme apyrase (5 units/mL), and the fibroblast growth factor (FGF) receptor inhibitor FIIN-2 (1 μM) decreased the frequency of spontaneous Ca²⁺ responses. These results indicate that ATP and FGF are involved in the spontaneous Ca²⁺ responses. SF2 cells differentiate into ameloblasts via interactions with mesenchymal cells. Therefore, SF2-G-GECO cells are expected to be a useful tool for studying the functions of Ca²⁺ responses in regulating gene expression during tooth development.