Anoctamin-6 Controls Bone Mineralization by Activating the Calcium Transporter NCX1

Anoctamin-6 (Ano6, TMEM16F) belongs to a family of putative Ca2+-activated Cl− channels and operates as membrane phospholipid scramblase. Deletion of Ano6 leads to reduced skeleton size, skeletal deformities, and mineralization defects in mice. However, it remains entirely unclear how a lack of Ano6 leads to a delay in bone mineralization by osteoblasts. The Na+/Ca2+ exchanger NCX1 was found to interact with Ano6 in a two-hybrid split-ubiquitin screen. Using human osteoblasts and osteoblasts from Ano6−/− and WT mice, we demonstrate that NCX1 requires Ano6 to efficiently translocate Ca2+ out of osteoblasts into the calcifying bone matrix. Ca2+-activated anion currents are missing in primary osteoblasts isolated from Ano6 null mice. Our findings demonstrate the importance of NCX1 for bone mineralization and explain why deletion of an ion channel leads to the observed mineralization defect: Ano6 Cl− currents are probably required to operate as a Cl− bypass channel, thereby compensating net Na+ charge movement by NCX1. Background: Deletion of Ano6 leads to reduced skeleton size and skeletal deformities. Results: Ca2+-activated anion currents are missing in primary osteoblasts from Ano6 null mice. Ano6 is required for the Na+/Ca2+ exchanger NCX1 to operate in osteoblasts. Conclusion: The Na+/Ca2+ exchanger NCX1 is essential for bone mineralization and requires Ano6 to fully operate. Significance: Ano6 supports NCX1, which is essential for bone mineralization.