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Journal of Neuroscience, Vol 15, 144-151, Copyright © 1995 by Society for Neuroscience
Cholinergic regulation of neurite outgrowth from isolated chick sympathetic neurons in culture
DH Small, G Reed, B Whitefield and V Nurcombe
Department of Pathology, University of Melbourne, Australia.
Neurotransmitters have been reported to regulate neurite outgrowth in
several vertebrate and nonvertebrate species. In this study, cultures of
isolated embryonic day 12 (E12) chick sympathetic neurons were grown in the
presence of cholinergic receptor agonists or antagonists. Both ACh and the
nonhydrolyzable cholinergic agonist carbamylcholine (CCh) inhibited neurite
outgrowth. ACh (0.1-1.0 mM) decreased the percentage of neurons bearing
neurites, but had no significant effect on cell survival. The effect of ACh
was increased in the presence of the cholinesterase inhibitors BW284C51 (1
microM), Tacrine (20 microM), and edrophonium (200 microM). Neurite
outgrowth was strongly inhibited by the muscarinic receptor agonist
oxotremorine (5-100 microM) and weakly inhibited by nicotine (50 nM to 10
microM). The inhibitory effect of CCh was decreased by the muscarinic
receptor antagonist atropine (10 microM), demonstrating that the effect of
CCh on neurite outgrowth was mediated, at least in part, through a
muscarinic receptor. The possibility that AChE can influence neurite
outgrowth directly, through a noncatalytic mechanism, was also examined.
When dissociated chick brain or sympathetic neurons were grown on plates
precoated with purified AChE, neurite outgrowth was strongly stimulated.
However, the neurite outgrowth-promoting effect of AChE was strictly
dependent upon the presence of substratum-bound heparan sulfate
proteoglycans (HSPG). Pretreatment of AChE with diisopropylfluorophosphate
to inhibit the esterase activity did not abolish this effect, suggesting
that the neurite outgrowth-promoting effect of AChE was associated with a
noncatalytic mechanism, a view supported by the observation that soluble
AChE had no effect on neurite outgrowth.(ABSTRACT TRUNCATED AT 250 WORDS)
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