Evidence That Calcium Entry Into Calcium-Transporting Dental Enamel Cells Is Regulated by Cholecystokinin,...

Dental enamel is formed by specialized epithelial cells which handle large quantities of Ca²⁺ while producing the most highly mineralized tissue. However, the mechanisms used by enamel cells to handle bulk Ca²⁺ safely remain unclear. Our previous work contradicted the dogma that Ca²⁺ is ferried through the cytosol of Ca²⁺-transporting cells and instead suggested an organelle-based route across enamel cells. This new paradigm involves endoplasmic reticulum (ER)-associated Ca²⁺ stores and their concomitant refilling by store-operated Ca²⁺ entry (SOCE) mediated by Ca²⁺ release activated Ca²⁺ (CRAC) channels. Given that Ca²⁺ handling is maximal during the enamel-mineralization stage (maturation), we anticipated that SOCE would also be elevated then. Confirmation was obtained here using single-cell recordings of cytosolic Ca²⁺ concentration ([Ca²⁺]_cyt) in rat ameloblasts. A candidate SOCE agonist, cholecystokinin (CCK), was found to be upregulated during maturation, with Cck transcript abundance reaching 30% of that in brain. CCK-receptor transcripts were also detected and Ca²⁺ imaging showed that CCK stimulation increased [Ca²⁺]_cyt in a dose-responsive manner that was sensitive to CRAC-channel inhibitors. Similar effects were observed with two other SOCE activators, acetylcholine and ATP, whose receptors were also found in enamel cells. These results provide the first evidence of a potential regulatory system for SOCE in enamel cells and so strengthen the Ca²⁺ transcytosis paradigm for ER-based transport of bulk Ca²⁺. Our findings also implicate enamel cells as a new physiological target of CCK and raise the possibility of an auto/paracrine system for regulating Ca²⁺ transport.