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Journal of Neuroscience, Vol 12, 1253-1261, Copyright © 1992 by Society for Neuroscience
Asymmetric modulation of cytosolic cAMP activity induces growth cone turning
AM Lohof, M Quillan, Y Dan and MM Poo
Department of Biological Sciences, Columbia University, New York, New York 10027.
The possible role of cyclic nucleotides as second messengers mediating
growth cone turning was studied by producing an asymmetric distribution of
cyclic nucleotides across the growth cone. A repetitive pulse application
method was developed to produce microscopic chemical gradients near the
growth cone of embryonic Xenopus neurons in cell culture. When picoliters
of a solution containing 20 mM dibutyryl cAMP (dB-cAMP), a
membrane-permeable analog of cAMP, were repetitively ejected from a
micropipette near the growth cone, neurite growth was consistently directed
toward the pipette. Theoretical analysis of the diffusion gradient showed
that the neurite is capable of detecting a 10% difference in dB-cAMP
concentration across the growth cone. Similar responses were also observed
using gradients of the phosphodiesterase inhibitor isobutylmethylxanthine,
or of forskolin, which activates adenylate cyclase. Dibutyryl cGMP,
however, produced no significant turning. These results suggest that a
cytoplasmic gradient of cAMP across the growth cone is sufficient to
initiate its turning response, and that cAMP in the growth cone could serve
as a second messenger in mediating the action of extracellular guidance
cues.
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