Regulation of MAP kinase by calcium-sensing receptor in bovine parathyroid and CaR-transfected HEK293 cells

Abstract

Regulation of the extracellular signal-regulated kinase 1 and 2 (ERK1/2) pathway by the extracellular calcium (Ca o 2+ )-sensing receptor (CaR) was investigated in bovine parathyroid and CaR-transfected human embryonic kidney (HEKCaR) cells. Elevating Ca o 2+ or adding the selective CaR activator NPS R-467 elicited rapid, dose-dependent phosphorylation of ERK1/2. These phosphorylations were attenuated by pretreatment with pertussis toxin (PTX) or by treatment with the phosphotyrosine kinase (PTK) inhibitors genistein and herbimycin, the phosphatidylinositol-specific phospholipase C (PI-PLC) inhibitor U-73122, or the protein kinase C (PKC) inhibitor GF109203X and were enhanced by the PKC activator phorbol 12-myristate 13-acetate. Combined treatment with PTX and inhibitors of both PKC and PTK nearly abolished high Ca o 2+ -evoked ERK1/2 activation in HEKCaR cells, demonstrating CaR-mediated coupling via both G q and G i . High Ca o 2+ increased serine phosphorylation of the 85-kDa cytosolic phospholipase A 2 (cPLA 2 ) in both parathyroid and HEKCaR cells. The selective mitogen-activated protein kinase (MAPK) inhibitor PD98059 abolished high-Ca o 2+ -induced ERK1/2 activation and reduced cPLA 2 phosphorylation in both cell types, documenting MAPK's role in cPLA 2 activation. Thus our data suggest that the CaR activates MAPK through PKC, presumably through G q/11 -mediated activation of PI-PLC, as well as through G i - and PTK-dependent pathway(s) in bovine parathyroid and HEKCaR cells and indicate the importance of MAPK in cPLA 2 activation.