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Journal of Neuroscience, Vol 12, 1160-1168, Copyright © 1992 by Society for Neuroscience
Selective activation of Ca(2+)-activated PKCs in Aplysia neurons by 5- HT
WS Sossin and JH Schwartz
Center for Neurobiology and Behavior, Columbia University, College of Physicians and Surgeons, New York, New York.
Ca(2+)-activated and Ca(2+)-independent protein kinase Cs (PKCs) are
present in the nervous system of the marine mollusk Aplysia californica
(Kruger et al., 1991). Sensitizing stimuli or application of the
facilitatory transmitter 5-HT to intact isolated ganglia produces the
presynaptic facilitation of sensory-to-motor neuron synapses that underlies
behavioral sensitization, which is a simple form of learning. Activation of
PKC can also produce this presynaptic facilitation (Braha et al., 1990). To
determine which type of PKC is activated, we developed a sensitive and
selective assay to measure both Ca(2+)- activated and Ca(2+)-independent
PKC activities in crude supernatant and membrane fractions of nervous
tissue. This assay is based on the specific binding of the Ca(2+)-activated
PKCs to phosphatidylserine vesicles in the presence of Ca2+ and makes use
of a novel synthetic peptide with sequences conforming to phylogenetically
conserved pseudosubstrate regions of the Ca(2+)-independent kinases. We
provide evidence that the presynaptic facilitation is produced by a Ca(2+)-
activated isoform: application of 5-HT increases the amount of the
Ca(2+)-activated PKC activity associated with the membrane. Under these
conditions, no increase in Ca(2+)-independent kinase activity is seen.
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