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Volume 17, Number 9, Issue of May 1, 1997 pp. 3085-3095
Copyright ©1997 Society for Neuroscience

Dynamic Microtubule Ends Are Required for Growth Cone Turning to Avoid an Inhibitory Guidance Cue

Received Nov. 7, 1996; revised Feb. 5, 1997; accepted Feb. 10, 1997.

Jean F. Challacombe, Diane M. Snow, and Paul C. Letourneau

Department of Cell Biology and Neuroanatomy, The University of Minnesota, Minneapolis, Minnesota 55455

Growth cone turning is an important mechanism for changing the direction of neurite elongation during development of the nervous system. Our previous study indicated that actin filament bundles at the leading margin direct the distal microtubular cytoskeleton as growth cones turn to avoid substratum-bound chondroitin sulfate proteoglycan. Here, we investigated the role of microtubule dynamics in growth cone turning by using low doses of vinblastine and taxol, treatments that reduce dynamic growth and shrinkage of microtubule ends. We used time-lapse phase-contrast videomicroscopy to observe embryonic chick dorsal root ganglion neuronal growth cones as they encountered a border between fibronectin and chondroitin sulfate proteoglycan in the presence and absence of 4 nM vinblastine or 7 nM taxol. Growth cones were fixed and immunocytochemically labeled to identify actin filaments and microtubules containing tyrosinated and detyrosinated -tubulin.

Our results show that after contact with substratum-bound chondroitin sulfate proteoglycan, vinblastine- and taxol-treated growth cones did not turn, as did controls; instead, they stopped or sidestepped. Even before drug-treated growth cones contacted a chondroitin sulfate proteoglycan border, they were narrower than controls, and the distal tyrosinated microtubules were less splayed and were closer to the leading edges of the growth cones. We conclude that the splayed dynamic distal ends of microtubules play a key role in the actin filament-mediated steering of growth cone microtubules to produce growth cone turning.

Key words: microtubule; growth cone; turning; actin filament; chondroitin sulfate proteoglycan; dynamic instability

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