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Differentiation of Mouse Embryonic Stem Cells into Cortical Interneuron Precursors
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Ependymal cell differentiation, from monociliated to multiciliated cells.

Nathalie Delgehyr1, Alice Meunier1, Marion Faucourt1

  • 1Institut de Biologie de l'Ecole Normal Supérieure (IBENS), Paris, France; INSERM, U1024, Paris, France; CNRS, UMR 8197, Paris, France.

Methods in Cell Biology
|April 4, 2015
PubMed
Summary

This study details methods for visualizing primary and motile cilia in developing brain ependymal cells. It provides protocols and marker lists for studying ciliary development and function.

Keywords:
Ependymal cellsMulticiliated cellsNeural cells primary cultureRadial glial cellsTubulin (poly)glutamylationTubulin (poly)glycylationTubulin detyrosinationWhole mount lateral ventricular walls

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Developmental Biology

Background:

  • Primary cilia are immotile signaling organelles in brain stem cells and neurons.
  • Motile cilia on ependymal cells drive cerebrospinal fluid flow.
  • Ependymal cell development offers a model for studying ciliation transitions.

Purpose of the Study:

  • To provide protocols for immunofluorescence staining of developing ependymal cells.
  • To identify markers for studying primary and motile cilia in the brain.
  • To investigate the transition from monociliated to multiciliated ependymal cells.

Main Methods:

  • Immunofluorescence staining of developing ependymal cells in vivo.
  • Whole-mount staining of lateral ventricle walls.
  • In vitro culture and staining of ependymal cells.
  • Utilizing markers for ciliary membrane proteins and axonemal tubulin modifications.

Main Results:

  • Established protocols for visualizing cilia in developing ependymal cells.
  • Compiled a list of validated markers for both primary and motile cilia.
  • Demonstrated methods applicable to both in vivo and in vitro systems.

Conclusions:

  • The provided protocols facilitate the study of ependymal cell differentiation and ciliation.
  • The marker list aids in characterizing ciliary structure and function during development.
  • This work supports research into cilia-related brain development and cerebrospinal fluid dynamics.