Chemical and physical properties of the extracellular matrix are required for the actin ring formation in osteoclasts

Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research
I NakamuraT Suda

Abstract

To examine the effect of extracellular matrix on osteoclast polarization, we focused on the actin organization in osteoclasts, using murine osteoclast-like multinucleated cells (OCLs) formed in cocultures of osteoblastic cells and bone marrow cells. When OCLs were cultured on either a plastic plate, calcified dentine, or calcium phosphate thin films in the presence of fetal bovine serum (FBS), they similarly formed ringed structures of F-actin dots (actin rings). However, OCLs placed on demineralized dentine or type I collagen gel matrix (collagen gel) failed to form actin rings. In the absence of FBS, actin ring formation in OCLs was induced on plastic plates coated with vitronectin, fibronectin, or type I collagen, but not on those coated with laminin, poly-L-lysine, or bovine serum albumin. Actin ring formation appeared to depend on integrins, since the GRGDS, but not the GRGES, peptide inhibited it in a dose-dependent manner. Moreover, immunoelectron microscopic examination revealed that vacuolar proton ATPase (V-ATPase) was localized along the apical membrane in much higher densities than the basolateral membrane in OCLs placed on plastic coverslips. In OCLs placed on collagen gel, however, V-ATPase was found to be distrib...Continue Reading

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