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A correction to this article has been published: J. Cell Biol. 144 (6) 1361
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© The Rockefeller University Press, 0021-9525/1999//447 $5.00
The Journal of Cell Biology, Volume 144, Number 3, , 1999 447-458

Regular Articles

Slow Axonal Transport of Neurofilament Protein in Cultured Neurons



Thomas J. Koehnle and Anthony Brown

Neuroscience Program, Department of Biological Sciences, Ohio University, Athens, Ohio 45701

We have investigated the axonal transport of neurofilament protein in cultured neurons by constricting single axons with fine glass fibers. We observed a rapid accumulation of anterogradely and retrogradely transported membranous organelles on both sides of the constrictions and a more gradual accumulation of neurofilament protein proximal to the constrictions. Neurofilament protein accumulation was dependent on the presence of metabolic substrates and was blocked by iodoacetate, which is an inhibitor of glycolysis. These data indicate that neurofilament protein moves anterogradely in these axons by a mechanism that is directly or indirectly dependent on nucleoside triphosphates. The average transport rate was estimated to be at least 130 µm/h (3.1 mm/d), and ~90% of the accumulated neurofilament protein remained in the axon after detergent extraction, suggesting that it was present in a polymerized form. Electron microscopy demonstrated that there were an abnormally large number of neurofilament polymers proximal to the constrictions. These data suggest that the neurofilament proteins were transported either as assembled polymers or in a nonpolymeric form that assembled locally at the site of accumulation. This study represents the first demonstration of the axonal transport of neurofilament protein in cultured neurons.

Key Words: neurofilament • cytoskeleton • slow axonal transport • axon • neuron

Abbreviations used in this paper: NF-L, low molecular weight neurofilament triplet polypeptide; PHEM, buffer containing Pipes, Hepes, EGTA, and MgCl2.

Address correspondence to Anthony Brown, Department of Biological Sciences, Ohio University, Athens, OH 45701. Tel.: (740) 593-2330. Fax: (740) 593-0300. E-mail: browna1{at}ohiou.edu

Funded by a grant from the National Institute of Neurological Disorders and Stroke.


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