title: Genetic manipulation of cerebellar granule neurons in vitro and in vivo to study neuronal morphology and migration.
abstract: Developmental events in the brain including neuronal morphogenesis and migration are highly orchestrated processes. In vitro and in vivo analyses allow for an in-depth characterization to identify pathways involved in these events. Cerebellar granule neurons (CGNs) that are derived from the developing cerebellum are an ideal model system that allows for morphological analyses. Here, we describe a method of how to genetically manipulate CGNs and how to study axono- and dendritogenesis of individual neurons. With this method the effects of RNA interference, overexpression or small molecules can be compared to control neurons. In addition, the rodent cerebellar cortex is an easily accessible in vivo system owing to its predominant postnatal development. We also present an in vivo electroporation technique to genetically manipulate the developing cerebella and describe subsequent cerebellar analyses to assess neuronal morphology and migration.
authors.affiliation: Cellular and Molelcular Neurobiology, Max Planck Institute of Experimental Medicine; Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB).
authors.forename: Anna
authors.initials: A
authors.lastname: Holubowska
authors.affiliation: Cellular and Molelcular Neurobiology, Max Planck Institute of Experimental Medicine; Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB).
authors.forename: Chaitali
authors.initials: C
authors.lastname: Mukherjee
authors.affiliation: Cellular and Molelcular Neurobiology, Max Planck Institute of Experimental Medicine; Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB).
authors.forename: Mayur
authors.initials: M
authors.lastname: Vadhvani
authors.affiliation: Cellular and Molelcular Neurobiology, Max Planck Institute of Experimental Medicine; Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB); stegmueller@em.mpg.de.
authors.forename: Judith
authors.initials: J
authors.lastname: Stegmüller
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology; D002465:Cell Movement / Q000502:physiology*; D002531:Cerebellum / Q000166:cytology / Q000502:physiology*; D003712:Dendrites / Q000502:physiology; D018274:Electroporation / Q000379:methods*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D051381:Rats; D014162:Transfection / Q000379:methods*