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The Journal of Neuroscience, July 27, 2005, 25(30):7014-7021; doi:10.1523/JNEUROSCI.2001-05.2005

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Cellular/Molecular
Neurofilament Polymer Transport in Axons

Yanping Yan1,2 and Anthony Brown1

1Center for Molecular Neurobiology and Department of Neuroscience, and 2Neuroscience Graduate Studies Program, The Ohio State University, Columbus, Ohio 43210

Neurofilament proteins are known to be transported along axons by slow axonal transport, but the form in which they move is controversial. In previous studies on cultured rat sympathetic neurons, we found that green fluorescent protein-tagged neurofilament proteins move predominantly in the form of filamentous structures, and we proposed that these structures are single-neurofilament polymers. In the present study, we have tested this hypothesis by using a rapid perfusion technique to capture these structures as they move through naturally occurring gaps in the axonal neurofilament array. Because the gaps lack neurofilaments, they permit unambiguous identification of the captured structure. Using quantitative immunofluorescence microscopy and correlative light and electron microscopy, we show that the captured structures are single continuous neurofilament polymers. Thus, neurofilament polymers are one of the cargo structures of slow axonal transport.

Key words: axonal transport (axoplasmic transport); cytoskeleton; neurofilament; ultrastructure; live-cell imaging; fluorescence microscopy

Received April 5, 2005; revised June 7, 2005; accepted June 13, 2005.




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