The Ca ²⁺-activated Cl ⁻ channel ANO1/TMEM16A regulates primary ciliogenesis

Abstract

The Ca ²⁺-activated Cl ⁻ channel ANO1/TMEM16A is located in the primary cilium, and blocking it pharmacologically or knocking it down with shRNA interferes with ciliogenesis. Before ciliogenesis, the channel is organized into a torus-shaped structure (the “nimbus”) enriched in proteins required for ciliogenesis.

Abstract

Many cells possess a single, nonmotile, primary cilium highly enriched in receptors and sensory transduction machinery that plays crucial roles in cellular morphogenesis. Although sensory transduction requires ion channels, relatively little is known about ion channels in the primary cilium (with the exception of TRPP2). Here we show that the Ca ²⁺-activated Cl ⁻ channel anoctamin-1 (ANO1/TMEM16A) is located in the primary cilium and that blocking its channel function pharmacologically or knocking it down with short hairpin RNA interferes with ciliogenesis. Before ciliogenesis, the channel becomes organized into a torus-shaped structure (“the nimbus”) enriched in proteins required for ciliogenesis, including the small GTPases Cdc42 and Arl13b and the exocyst complex component Sec6. The nimbus excludes F-actin and coincides with a ring of acetylated microtubules. The nimbus appears to form before, or independent of, apical docking of the mother centriole. Our data support a model in which the nimbus provides a scaffold for staging of ciliary components for assembly very early in ciliogenesis and chloride transport by ANO1/TMEM16A is required for the genesis or maintenance of primary cilia.