title: Visualization of Clathrin-Mediated Endocytosis During Semaphorin-Guided Axonal Growth.
abstract: Semaphorin3A (Sema3A) guides axonal growth during neuronal network development. Accumulating evidence indicates that Sema3A-induced growth cone collapse and repulsion involve endocytic membrane trafficking in the growth cone. It is now possible to visualize endocytic processes in living cells using total internal reflection fluorescence microscopy (TIRFM), a powerful tool for imaging dynamic subcellular events at the plasma membrane. In this chapter, we describe a method for TIRFM observation and analysis of clathrin-mediated endocytosis in growth cones of chicken dorsal root ganglion neurons that receive an extracellular concentration gradient of Sema3A in a culture medium.
authors: Laboratory for Neuronal Growth Mechanisms, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
authors: Rurika
authors: R
authors: Itofusa
authors: Laboratory for Neuronal Growth Mechanisms, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.|PRESTO, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012, Japan.
authors: Takuro
authors: T
authors: Tojima
authors: Laboratory for Neuronal Growth Mechanisms, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan. kamiguchi@brain.riken.jp.
authors: Hiroyuki
authors: H
authors: Kamiguchi
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology*; D002642:Chick Embryo; D002966:Clathrin / Q000502:physiology*; D004705:Endocytosis / Q000502:physiology*; D005727:Ganglia, Spinal / Q000166:cytology; D020439:Growth Cones / Q000502:physiology; D008856:Microscopy, Fluorescence / Q000379:methods; D040121:Semaphorin-3A / Q000502:physiology*; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D002966:Clathrin; D040121:Semaphorin-3A
keywords: Axon guidance; Clathrin; Dorsal root ganglion neuron; Dynamin 1; Endocytosis; Growth cone; Semaphorin3A; Total internal reflection fluorescence microscopy