Using photoactivatable GFP to track axonal transport kinetics.
The advent of photoactivatable tools has revolutionized imaging of dynamic cellular processes. One such application is to visualize axonal transport-an intricate and dynamic process by which proteins and other macromolecules are conveyed from their sites of synthesis in the cell bodies to their destinations within axons and synapses. High-quality dynamic imaging of axonal transport using photoactivatable vectors can now be routinely performed using epifluorescence microscopes and CCD cameras that are standard in most laboratories, yet this is largely underutilized. Here we describe detailed protocols for imaging cargoes moving in fast and slow axonal transport in axons of cultured hippocampal neurons.
Department of Pathology, University of California, San Diego, 9500 Gilman Drive, BSB 1029, MC0612, La Jolla, CA, 92093, USA, arganguly@ucsd.edu.
Archan
A
Ganguly
Subhojit
S
Roy
D000818:Animals; D001370:Axonal Transport; D001369:Axons / Q000378:metabolism*; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology; D007700:Kinetics; D051379:Mice; D008856:Microscopy, Fluorescence; D055668:Photochemical Processes; D021381:Protein Transport; D059010:Single-Cell Analysis / Q000379:methods
D049452:Green Fluorescent Proteins