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Journal of Neuroscience, Vol 2, 1673-1681, Copyright © 1982 by Society for Neuroscience
Inhibitor of adenosine 3':5'-monophosphate-dependent protein kinase blocks presynaptic facilitation in Aplysia
VF Castellucci, A Nairn, P Greengard, JH Schwartz and ER Kandel
Sensitization of the gill withdrawal reflex results from presynaptic
facilitation at the excitatory synapses made by sensory neurons on gill
motor neurons. Facilitation is accompanied by an increase in the duration
of the action potential in sensory cells because of the depression of a K+
current. This results in an increasd influx of CA2+ and a greater release
of transmitter from sensory neurons. There is evidence that serotonin is
the facilitating transmitter and that the depression of the K+ current by
serotonin mediated by cAMP-dependent protein phosphorylation. To test
further the role of the cAMP-dependent protein kinase and of protein
phosphorylation in sensitization, we have attempted to prevent or reverse
the development of the electrophysiological correlates that accompany
sensitization. We have pressure-injected sensory neurons with a specific
and a stable protein inhibitor of the cAMP-dependent protein kinase both
before and after the application of serotonin or the activation of the
facilitator neurons. The increase in spike broadening that accompanies
facilitation was prevented or diminished by injection of the inhibitor.
Moreover, injection of the inhibitor could reverse fully the developed
spike broadening produced by prior application of serotonin. These
observations strenthen the evidence for the involvement of protein
phosphorylation in presynaptic facilitation. Phosphorylation of the
substrate protein evidently is quite labile and does not persist after the
kinase is inhibited. Thus, the time course of short term sensitization
appears to be determined by an active kinase. We think that it is likely
that the mechanism for maintaining the kinase in an active form resides in
the slow decay of the cAMP produced by the action of serotonin or the
facilitator neurons on the sensory cells.
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