Live-Imaging Centriole Amplification in Mouse Brain Multiciliated Cells.

Multiciliated cells (MCC) display on their apical surface hundreds of beating cilia that propel physiological fluids. They line brain ventricles where they propel the cerebrospinal liquid, airways where they clear mucus and pathogens and reproductive ducts where they concentrate the sperm in males or drive the egg along the oviducts in females. Motile cilia are nucleated from basal bodies which are modified centrioles. MCC therefore evade centriole archetypal duplication program to make several hundreds and nucleate an identical number of motile cilia. Defects in this centriole amplification process lead to severe human pathologies called "ciliary aplasia" or "acilia syndrome" and more recently renamed "reduced generation of motile cilia" (RGMC). Patients with this syndrome present frequent hydrocephaly, lung failure, and subfertility. In this manuscript, we describe the protocol we developed and optimized over the years to live image the centriole amplification dynamics. We explain why mouse brain MCC is a good model and provide the tips to enable successful spatially and temporally resolved monitoring of this massive organelle reorganization.