TY - JOUR TI - In vitro Time-lapse Imaging of Primary Cilium in Migrating Neuroblasts AU - Sawada, Masato AU - Matsumoto, Mami AU - Narita, Keishi AU - Kumamoto, Natsuko AU - Ugawa, Shinya AU - Takeda, Sen AU - Sawamoto, Kazunobu VL - 10 IS - 22 PY - 2020 DA - 2020/11/20 SP - e3823 C1 - Bio-protocol 2020;10:e3823 DO - 10.21769/BioProtoc.3823 UR - https://doi.org/10.21769/BioProtoc.3823 AB - Neuronal migration is a critical step for the development of neuronal circuits in the brain. Immature new neurons (neuroblasts) generated in the postnatal ventricular-subventricular zone (V-SVZ) show a remarkable potential to migrate for a long distance at a high speed in the postnatal mammalian brain, and are thus a powerful model to analyze the molecular and cellular mechanisms of neuronal migration. Here we describe a methodology for in vitro time-lapse imaging of the primary cilium and its related structures in migrating V-SVZ-derived neuroblasts using confocal or superresolution laser-scanning microscopy. The V-SVZ tissues are dissected from postnatal day 0-1 (P0-1) mouse brains and dissociated into single cells by trypsinization and gentle pipetting. These cells are then transduced with a plasmid(s) encoding a gene(s) of interest, aggregated by centrifugation, and cultured for 2 days in Matrigel. Time-lapse images of migratory behaviors of cultured neuroblasts and their ciliary structures, including the ciliary membrane and basal body, are acquired by confocal or superresolution laser-scanning microscopy. This method provides information about the spatiotemporal dynamics of neuroblasts’ morphology and ciliary structures, and is widely applicable to various types of migrating neuronal and nonneuronal cells in various species. KW - Postnatal neurogenesis KW - Ventricular-subventricular zone KW - Neuroblast KW - Primary cilium KW - Migration KW - Time-lapse imaging KW - Confocal microscopy KW - Superresolution microscopy JF - Bio-protocol SN - 2331-8325 PB - Bio-protocol LLC. BIO101 - False