title: Axonal actin in action: Imaging actin dynamics in neurons.
abstract: Actin is a highly conserved, key cytoskeletal protein involved in numerous structural and functional roles. In neurons, actin has been intensively investigated in axon terminals-growth cones-and dendritic spines, but details about actin structure and dynamics in axon shafts have remained obscure for decades. A major barrier in the field has been imaging actin. Actin exists as soluble monomers (G-actin) as well as actin filaments (F-actin), and labeling actin with conventional fluorescent probes like GFP/RFP typically leads to a diffuse haze that makes it difficult to discern kinetic behaviors. In a recent publication, we used F-actin selective probes to visualize actin dynamics in axons, resolving striking actin behaviors that have not been described before. However, using these probes to visualize actin dynamics is challenging as they can cause bundling of actin filaments; thus, experimental parameters need to be strictly optimized. Here we describe some practical methodological details related to using these probes for visualizing F-actin dynamics in axons.
authors.affiliation: Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA.
authors.forename: Kelsey
authors.initials: K
authors.lastname: Ladt
authors.affiliation: Department of Pathology, University of California, San Diego, La Jolla, CA, USA.
authors.forename: Archan
authors.initials: A
authors.lastname: Ganguly
authors.affiliation: Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA; Department of Pathology, University of California, San Diego, La Jolla, CA, USA.
authors.forename: Subhojit
authors.initials: S
authors.lastname: Roy
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001369:Axons / Q000378:metabolism*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D003712:Dendrites / Q000502:physiology; D006624:Hippocampus / Q000166:cytology*; D007734:Kymography / Q000379:methods; D051379:Mice; D008856:Microscopy, Fluorescence; D013194:Staining and Labeling; D014162:Transfection
chemical_list: D000199:Actins
keywords: Actin; F-actin probes; Lifeact; Utrophin