Live-cell imaging of neurofilament transport in cultured neurons. Methods Cell Biol 2016;131:21-90
Date
01/23/2016Pubmed ID
26794508Pubmed Central ID
PMC6007976DOI
10.1016/bs.mcb.2015.07.001Scopus ID
2-s2.0-84940688867 (requires institutional sign-in at Scopus site) 9 CitationsAbstract
Neurofilaments, which are the intermediate filaments of nerve cells, are space-filling cytoskeletal polymers that contribute to the growth of axonal caliber. In addition to their structural role, neurofilaments are cargos of axonal transport that move along microtubule tracks in a rapid, intermittent, and bidirectional manner. Though they measure just 10nm in diameter, which is well below the diffraction limit of optical microscopes, these polymers can reach 100 μm or more in length and are often packed densely, just tens of nanometers apart. These properties of neurofilaments present unique challenges for studies on their movement. In this article, we describe several live-cell fluorescence imaging strategies that we have developed to image neurofilament transport in axons of cultured neurons on short and long timescales. Together, these methods form a powerful set of complementary tools with which to study the axonal transport of these unique intracellular cargos.
Author List
Uchida A, Monsma PC, Fenn JD, Brown AMESH terms used to index this publication - Major topics in bold
AnimalsAxonal Transport
Axons
Cell Culture Techniques
Cells, Cultured
Cerebral Cortex
Cytoskeleton
Ganglia, Spinal
Intermediate Filaments
Kymography
Mice
Microscopy, Fluorescence
Microtubules
Rats
Recombinant Fusion Proteins
Software
Staining and Labeling
Superior Cervical Ganglion
Transfection
