Ciliated cell domains with locally coordinated ciliary motion generate a mosaic of microflows in the brain's lateral ventricles.

Circulation of cerebrospinal fluid (CSF) through the brain's ventricles is essential for maintaining brain homeostasis and supporting neurogenesis. CSF flow is supported by the structural polarization of multiciliated cells, which align with the flow direction. However, it remains unclear how the organization of tissue-wide polarity across the ciliary epithelium comprised of thousands of cells, determines the trajectory of the flow and efficient distribution of the CSF. Here, we used new approaches to analyze the organization of translational polarity across extensive areas of the lateral ventricular wall. We also used live imaging to examine cilia motion, flow trajectories, and ciliary beat frequency (CBF) in live preparations of ventricles. In addition to the primary flow running across the ventricular wall from the posterior area to the anterior (P-A), we found multiple local microflows with both direct and curved trajectories that deviate from the mainstream P-A direction. Our results suggest that the ciliated epithelium in the lateral ventricles varies in the alignment of ciliated cell translational polarity: whereas in the narrow dorsal area translational polarity is aligned with the direction of the mainstream flow, in the periphery of the mainstream it is organized into distinct cell clusters with locally aligned polarity vectors. We posit that the cluster organization of the multiciliated ependymal cells underpins the generation of a complex mosaic of flows, with the local microflows facilitating the wide spreading of the CSF across the ependyma. We demonstrate that nNOS is involved in the control of translational polarity, cluster organization, microflows, and CBF in the ependyma.