Calcitonin induces podosome disassembly and detachment of osteoclasts by modulating Pyk2 and Src activities

Abstract: Osteoclasts (OCs) attach to the extracellular matrix via specialized attachment structures called podosomes, which form a prominent F-actin-rich ring that is thought to correspond to the sealing zone of resorbing OCs. Calcitonin (CT), a 32-amino acid polypeptide, inhibits bone resorption by decreasing motility, inducing retraction, disassembling podosome, and disrupting the actin-ring structure of OCs. However, the detailed mechanisms of how CT induces the disassembly of podosome and disruption of the adhesive structures in OCs are not well characterized. Pyk2 localizes in the sealing zone of OCs. It is activated by ligation of integrins, and then activates Src, an important signaling molecule for bone resorption. Thus, the Pyk2/Src complex in podosome could be a potential target for the CT-induced signaling. Using interference reflection, phase contrast, and confocal microscopy, CT effects on the dynamic changes of peripheral adhesive structure in living OCs were examined. CT induced dephosphorylation at Tyr402 of Pyk2 and decreased its labeling at peripheral adhesion region, which would prevent formation of the Pyk2/Src complex in this region. CT induced increase of intracellular phosphorylation of Tyr402 Pyk2 and increase of dephosphorylation at Tyr527 of Src and Pyk2/Src colocalization in the central region of OCs. This evidence suggested that Src might function as an adaptor protein that competes for Pyk2 and relocates it from peripheral adhesive zone to the central region of OCs. In conclusion, CT may induce podosome reassembly and peripheral adhesive zone detachment by modulating Pyk2 and Src phosphorylation state and their intracellular distribution in OCs. [Copyright &y& Elsevier]