Sensitivity of Ca2+-sensing receptor-transient receptor potential-mediated Ca2+ influx to extracellular acidity in bEND.3 endothelial cells.

Abstract

Ca²⁺-sensing receptors (CaSRs) are G protein-coupled receptors activated by elevated concentrations of extracellular Ca²⁺. In our previous works, we showed protein and functional expression of CaSR in mouse cerebral endothelial cell (EC) (bEND.3); the CaSR response (high Ca²⁺-elicited cytosolic [Ca²⁺] elevation) was unaffected by suppression of phospholipase C but in part involved Ca²⁺ influx through transient receptor potential V1 (TRPV1) channels. In this work, we investigated if extracellular acidity affected CaSR-mediated Ca²⁺ influx triggered by high (3 mM) Ca²⁺ (CaSR agonist), 3 mM spermine (CaSR agonist), and 10 mM cinacalcet (positive allosteric modulator of CaSR). Extracellular acidosis (pH 6.8 and pH 6.0) strongly suppressed cytosolic [Ca²⁺] elevation triggered by high Ca²⁺, spermine, and cinacalcet; acidosis also inhibited Mn²⁺ influx stimulated by high Ca²⁺ and cinacalcet. Purinoceptor-triggered Ca²⁺ response, however, was not suppressed by acidosis. Extracellular acidity also did not affect membrane potential, suggesting suppressed CaSR-mediated Ca²⁺ influx in acidity did not result from the reduced electrical driving force for Ca²⁺. Our results suggest Ca²⁺ influx through a putative CaSR-TRP complex in bEND.3 EC was sensitive to extracellular pH.