The preparation of primary cortical neuron cultures and a practical application using immunofluorescent cytochemistry.
Traditionally, cultures of primary cortical neurons are prepared from embryonic animals because at prenatal stages neurons have not yet developed extensive axonal and dendritic arbors and are not highly innervated, thus rendering the cells less susceptible to damage during dissociation of the neuronal tissue. The appropriate developmental age for preparing primary cultures of any cell type is determined by the time at which the cells of interest are generated and abundant. Most cerebral cortical neurons are generated between embryonic days (E) 11 and 17 in the mouse (embryos being considered 0.5 days old when a vaginal plug is detected in the morning). Here we describe a method to obtain short-term cultures of mouse primary cortical neurons at E15.5 and a practical application using fluorescent immunocytochemistry.
Sciarretta
Carla
C
Mouse Biology Unit, European Molecular Biology Laboratory, Monterotondo 00015, Italy.
Minichiello
Liliana
L
D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002540:Cerebral Cortex / Q000166:cytology*; D004210:Dissection; D004622:Embryo, Mammalian; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects
D019208:Brain-Derived Neurotrophic Factor; D020928:Mitogen-Activated Protein Kinases