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The Journal of Neuroscience, March 15, 1999, 19(6):1965-1975
Myelin and Collapsin-1 Induce Motor Neuron Growth Cone Collapse
through Different Pathways: Inhibition of Collapse by Opposing Mutants
of Rac1
Thomas B.
Kuhn1,
Michael D.
Brown2,
Christine L.
Wilcox3,
Jonathan A.
Raper4, and
James R.
Bamburg1
Departments of 1 Biochemistry and Molecular Biology,
2 Anatomy and Neurobiology, and 3 Microbiology,
Colorado State University, Fort Collins, Colorado 80523, and
4 Department of Neurosciences, University of Pennsylvania,
School of Medicine, Philadelphia, Pennsylvania 19104
Precise growth cone guidance is the consequence of a continuous
reorganization of actin filament structures within filopodia and
lamellipodia in response to inhibitory and promoting cues. The small
GTPases rac1, cdc42, and rhoA are critical for regulating distinct
actin structures in non-neuronal cells and presumably in growth cones.
Collapse, a retraction of filopodia and lamellipodia, is a typical
growth cone behavior on contact with inhibitory cues and is associated
with depolymerization and redistribution of actin filaments. We
examined whether small GTPases mediate the inhibitory properties of CNS
myelin or collapsin-1, a soluble semaphorin, in chick embryonic motor
neuron cultures. As demonstrated for collapsin-1, CNS myelin-evoked
growth cone collapse was accompanied by a reduction of
rhodamine-phalloidin staining most prominent in the growth cone
periphery, suggesting actin filament disassembly. Specific mutants of
small GTPases were capable of desensitizing growth cones to CNS myelin
or collapsin-1. Adenoviral-mediated expression of constitutively active
rac1 or rhoA abolished CNS myelin-induced collapse and allowed
remarkable neurite extension on a CNS myelin substrate. In contrast,
expression of dominant negative rac1 or cdc42 negated
collapsin-1-induced growth cone collapse and promoted neurite outgrowth
on a collapsin-1 substrate. These findings suggest that small GTPases
can modulate the signaling pathways of inhibitory stimuli and,
consequently, allow the manipulation of growth cone behavior. However,
the fact that opposite mutants of rac1 were effective against different
inhibitory stimuli speaks against a universal signaling pathway
underlying growth cone collapse.
Key words:
growth cone; actin filaments; small GTPases; myelin; collapsin-1; rac1; semaphorin
Copyright © 1999 Society for Neuroscience 0270-6474/99/1961965-11$05.00/0
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