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Abstract

The specific phenotypes and progression to maturity of primary cortical neurons
in long-term culture correlate well with neurons in vivo. Utilizing a model of
neuronal injury in long-term cultures at 21 days in vitro (DIV), we have identified
a distinct population of neurons that translocate into the injury site. 5-
Bromo-20-deoxyUridine (BrdU) incorporation studies demonstrated that neurons
with the capacity to translocate were 21 days old. However, this motile ability is
not consistent with the traditional view of the maturation and structural stability
of neurons in long-term culture. Therefore, we examined the neurons’ cytoskeletal
profile using immunocytochemistry, to establish relative stage of maturation
and phenotype. Expression of marker proteins including b-III-tubulin, a-internexin,
NF-L and NF-M, tau and L1 indicated the neurons were differentiated,
and in some cases polarized. The neurons did not immunolabel with NF-H or
MAP2, which might suggest they had not reached the level of maturity of other
neurons in culture. They did not express the microtubule-associated migration
marker doublecortin (DCX). Cytoskeletal disrupting agents were used to further
investigate the role of the microtubule cytoskeleton in translocation, and microtubule
destabilization significantly enhanced aspects of their motility. Finally,
molecular guidance cues affected their motility in a similar manner to that
reported for both axon guidance and early neuron migration. Therefore, this
study has identified and characterized a population of motile neurons in vitro that
have the capacity to migrate into a site of injury. These studies provide new information
on the structurally dynamic features of subsets of neurons.

Item Type:	Article
Authors/Creators:	Haas, MA and Chuckowree, JA and Chung, RS and Vickers, JC and Dickson, TC
Keywords:	cytoskeleton; migration; taxol; nocodazole; molecular guidance cues
Journal or Publication Title:	Cell Motility and the Cytoskeleton
Publisher:	John Wiley & Sons, Inc.
ISSN:	0886-1544
DOI / ID Number:	https://doi.org/10.1002/cm.20182
Additional Information:	The original publication is available at http://www.interscience.wiley.com/
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