title: Ex vivo imaging of postnatal cerebellar granule cell migration using confocal macroscopy.
abstract: During postnatal development, immature granule cells (excitatory interneurons) exhibit tangential migration in the external granular layer, and then radial migration in the molecular layer and the Purkinje cell layer to reach the internal granular layer of the cerebellar cortex. Default in migratory processes induces either cell death or misplacement of the neurons, leading to deficits in diverse cerebellar functions. Centripetal granule cell migration involves several mechanisms, such as chemotaxis and extracellular matrix degradation, to guide the cells towards their final position, but the factors that regulate cell migration in each cortical layer are only partially known. In our method, acute cerebellar slices are prepared from P10 rats, granule cells are labeled with a fluorescent cytoplasmic marker and tissues are cultured on membrane inserts from 4 to 10 hr before starting real-time monitoring of cell migration by confocal macroscopy at 37 °C in the presence of CO2. During their migration in the different cortical layers of the cerebellum, granule cells can be exposed to neuropeptide agonists or antagonists, protease inhibitors, blockers of intracellular effectors or even toxic substances such as alcohol or methylmercury to investigate their possible role in the regulation of neuronal migration.
authors.affiliation: PRIMACEN, Cell Imaging Platform of Normandy, Inserm, IRIB, University of Rouen.
authors.forename: Magalie
authors.initials: M
authors.lastname: Bénard
authors.affiliation: PRIMACEN, Cell Imaging Platform of Normandy, Inserm, IRIB, University of Rouen.
authors.forename: Alexis
authors.initials: A
authors.lastname: Lebon
authors.affiliation: Department of Neurobiology, School of Medicine, Yale University.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Komuro
authors.affiliation: PRIMACEN, Cell Imaging Platform of Normandy, Inserm, IRIB, University of Rouen.
authors.forename: David
authors.initials: D
authors.lastname: Vaudry
authors.affiliation: PRIMACEN, Cell Imaging Platform of Normandy, Inserm, IRIB, University of Rouen; ludovic.galas@univ-rouen.fr.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Galas
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000502:physiology*; D002525:Cerebellar Cortex / Q000166:cytology; D002531:Cerebellum / Q000166:cytology*; D003594:Cytoplasmic Granules; D005260:Female; D007395:Interneurons / Q000166:cytology; D008297:Male; D018613:Microscopy, Confocal / Q000379:methods*; D009417:Nerve Tissue / Q000166:cytology; D009474:Neurons / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar