Bone-resorbing osteoclasts contain gap-junctional connexin-43

Abstract

Intercellular gap junctions have been previously described at contact sites between surface osteoblasts, between osteoblasts and underlying osteocytes, and between osteocyte cell processes in the canaliculi. The subunits of gap junction channels are assembled from a family of proteins called connexins. In the present work, we show that rat osteoclasts cultured on bovine bone slices show connexin-43 (Cx43) staining localizing in the plasma membrane of the cells in cell-cell contacts and over the basolateral membrane of osteoclasts. The effect of heptanol, a known gap-junctional inhibitor, was studied using the well-characterized pit formation assay. Heptanol decreased the number and activity of osteoclasts. The proportion of mononuclear tartrate-resistant acid phosphatase (TRAP)-positive cells out of all TRAP-positive cells increased on heptanol treatment, suggesting a defect in the fusion of mononuclear osteoclast precursors to multinucleated mature osteoclasts. Furthermore, the total resorbed area and the number of resorption pits also decreased in the heptanol-treated cultures. These results suggest that gap-junctional Cx43 plays a functional role in osteoclasts and that the blocking of gap junctions decreases both the number and the activity of osteoclasts. This can indicate both a direct communication between multinucleated osteoclasts and mononuclear cells through gap junctions or an indirect effect through gap junctions between osteoblasts.