title: Culturing of cerebellar granule cells to study neuronal migration: gradient and local perfusion assays.
abstract: Cultures of cerebellar granule cells are a suitable model to analyze the mechanisms governing neuronal migration. In this unit, we describe a protocol to obtain cultures of dissociated granule cells at a low density, where individual cells can be easily observed. In addition, we include a protocol for studying neuronal migration in these cultures, using single, actively migrating cerebellar granule cells. Following this protocol, a factor of interest can be applied either in a gradient concentration by means of a micropipet located near the neuron, or in a homogeneous concentration by locally perfusing a certain region of the neuron. Time-lapse images are taken to analyze changes in the speed and/or directionality of the observed neuron. Overall, the two protocols take more or less a day and a half to perform, and are a useful way to evaluate a certain factor/drug for its chemotactic activity or its capacity to alter migration speed.
authors: CAS-MPG Partner Institute for Computational Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors: Patricia
authors: P
authors: Guijarro
authors: Jian
authors: J
authors: Jiang
authors: Xiao-bing
authors: XB
authors: Yuan
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002465:Cell Movement / Q000502:physiology*; D002531:Cerebellum / Q000166:cytology*; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D009474:Neurons / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D017077:Culture Media, Conditioned