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Published online February 11, 2008
doi:10.1083/jcb.200707042
The Journal of Cell Biology, Vol. 180, No. 3, 619-632
The Rockefeller University Press, 0021-9525 $30.00
© 2008 Witte et al.
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Article

 Microtubule stabilization specifies initial neuronal polarization

Harald Witte, Dorothee Neukirchen, and Frank Bradke

Axonal Growth and Regeneration, Max Planck Institute of Neurobiology, 82152 Martinsried, Germany

Correspondence to F. Bradke: fbradke{at}neuro.mpg.de

Axon formation is the initial step in establishing neuronal polarity. We examine here the role of microtubule dynamics in neuronal polarization using hippocampal neurons in culture. We see increased microtubule stability along the shaft in a single neurite before axon formation and in the axon of morphologically polarized cells. Loss of polarity or formation of multiple axons after manipulation of neuronal polarity regulators, synapses of amphids defective (SAD) kinases, and glycogen synthase kinase-3β correlates with characteristic changes in microtubule turnover. Consistently, changing the microtubule dynamics is sufficient to alter neuronal polarization. Application of low doses of the microtubule-destabilizing drug nocodazole selectively reduces the formation of future dendrites. Conversely, low doses of the microtubule-stabilizing drug taxol shift polymerizing microtubules from neurite shafts to process tips and lead to the formation of multiple axons. Finally, local stabilization of microtubules using a photoactivatable analogue of taxol induces axon formation from the activated area. Thus, local microtubule stabilization in one neurite is a physiological signal specifying neuronal polarization.

Abbreviations used in this paper: ANOVA, analysis of variance; CRMP-2, collapsin response mediator protein 2; DIV, days in vitro; EB3, end-binding protein 3; GSK-3β, glycogen synthase kinase-3β; MAP, microtubule-associated protein; MT, microtubule; PAR-1, partitioning defective-1; PI3K, phosphoinositide 3-kinase; SAD, synapses of amphids defective; TSA, trichostatin A.


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