A RYANODINE RECEPTOR-LIKE MOLECULE EXPRESSED IN THE OSTEOCLAST PLASMA-MEMBRANE FUNCTIONS IN EXTRACELLULAR CA2+

Ryanodine receptors (RyRs) reside in microsomal membranes where they gate Ca2+ release in response to changes in the cytosolic Ca2+ concentration. In the osteoclast, a divalent cation sensor, the Ca2+ receptor (CaR), located within the cell's plasma membrane, monitors changes in the extracellular Ca2+ concentration. Here we show that a RyR-like molecule is a functional component of this receptor, We have demonstrated that [H-3] ryanodine specifically binds to freshly isolated rat osteoclasts, The binding was displaced by ryanodine itself, the CaR agonist Ni2+ and the RyR antagonist ruthenium red, The latter also inhibited cytosolic Ca2+ elevations induced by Ni2+, In contrast, the responses to Ni2+ were strongly potentiated by an antiserum Ab(129) raised to an epitope located within the channel-forming domain of the type II RyR The antiserum also stained the surface of intact, unfixed, trypan blue-negative osteoclasts, Serial confocal sections and immunogold scanning electron microscopy confirmed a plasma membrane localization of this shining, Antiserum Ab(34) directed to a putatively intracellular RyR epitope expectedly did not stain live osteoclasts nor did it potentiate CaR activation, It did, however, stain fixed, permeabilized cells in a distinctive cytoplasmic pattern, We conclude that an RyR-like molecule resides within the osteoclast plasma membrane and plays an important role in extracellular Ca2+ sensing.