title: Ependymal cell differentiation, from monociliated to multiciliated cells.
abstract: Primary and motile cilia differ in their structure, composition, and function. In the brain, primary cilia are immotile signalling organelles present on neural stem cells and neurons. Multiple motile cilia are found on the surface of ependymal cells in all brain ventricles, where they contribute to the flow of cerebrospinal fluid. During development, monociliated ependymal progenitor cells differentiate into multiciliated ependymal cells, thus providing a simple system for studying the transition between these two stages. In this chapter, we provide protocols for immunofluorescence staining of developing ependymal cells in vivo, on whole mounts of lateral ventricle walls, and in vitro, on cultured ependymal cells. We also provide a list of markers we currently use to stain both types of cilia, including proteins at the ciliary membrane and tubulin posttranslational modifications of the axoneme.
authors.affiliation: Institut de Biologie de l'Ecole Normal Supérieure (IBENS), Paris, France; INSERM, U1024, Paris, France; CNRS, UMR 8197, Paris, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Delgehyr
authors.affiliation: Institut de Biologie de l'Ecole Normal Supérieure (IBENS), Paris, France; INSERM, U1024, Paris, France; CNRS, UMR 8197, Paris, France.
authors.forename: Alice
authors.initials: A
authors.lastname: Meunier
authors.affiliation: Institut de Biologie de l'Ecole Normal Supérieure (IBENS), Paris, France; INSERM, U1024, Paris, France; CNRS, UMR 8197, Paris, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Faucourt
authors.affiliation: Curie Institute, Orsay, France; CNRS, UMR3306, Orsay, France; INSERM, U1005, Orsay, France; INSERM, UMRS 1120; Unité de Génétique et Physiologie de l'Audition, Pasteur Institute, Paris, France.
authors.forename: Montserrat
authors.initials: M
authors.lastname: Bosch Grau
authors.affiliation: Institut de Biologie de l'Ecole Normal Supérieure (IBENS), Paris, France; INSERM, U1024, Paris, France; CNRS, UMR 8197, Paris, France; INSERM, UMRS 975, Brain and Spinal Cord Institute, Hôpital de la Salpêtrière, Paris, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Strehl
authors.affiliation: Curie Institute, Orsay, France; CNRS, UMR3306, Orsay, France; INSERM, U1005, Orsay, France.
authors.forename: Carsten
authors.initials: C
authors.lastname: Janke
authors.affiliation: Institut de Biologie de l'Ecole Normal Supérieure (IBENS), Paris, France; INSERM, U1024, Paris, France; CNRS, UMR 8197, Paris, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Spassky
mesh_terms: D000071916:AC133 Antigen; D020727:ADP-Ribosylation Factors / Q000502:physiology; D000262:Adenylyl Cyclases / Q000502:physiology; D000818:Animals; D015703:Antigens, CD; D015415:Biomarkers; D051927:CD24 Antigen; D002454:Cell Differentiation; D002478:Cells, Cultured; D002923:Cilia / Q000502:physiology*; D004805:Ependyma / Q000166:cytology* / Q000502:physiology / Q000601:surgery; D063928:Ependymoglial Cells / Q000166:cytology*; D006023:Glycoproteins; D007150:Immunohistochemistry; D020547:Lateral Ventricles / Q000166:cytology* / Q000502:physiology / Q000601:surgery; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology*; D010455:Peptides; D061251:Primary Cell Culture / Q000379:methods; D013194:Staining and Labeling / Q000379:methods; D014404:Tubulin / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; C560093:Arl13b protein, mouse; D015415:Biomarkers; D051927:CD24 Antigen; C497890:Cd24a protein, mouse; D006023:Glycoproteins; D010455:Peptides; D014404:Tubulin; D020727:ADP-Ribosylation Factors; D000262:Adenylyl Cyclases; C421784:adenylate cyclase 3
keywords: Ependymal cells; Multiciliated cells; Neural cells primary culture; Radial glial cells; Tubulin (poly)glutamylation; Tubulin (poly)glycylation; Tubulin detyrosination; Whole mount lateral ventricular walls