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J Neurophysiol 97: 134-143, 2007. First published September 13, 2006; doi:10.1152/jn.00122.2006
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PKC Modulation of Transmitter Release by SNAP-25 at Sensory-to-Motor Synapses in Aplysia

Gry Houeland1, Arash Nakhost2, Wayne S. Sossin2 and Vincent F. Castellucci1

1Department of Physiology, University of Montreal; and 2Department of Neurology and Neurosurgery, McGill University, Montreal Neurological Institute, Montreal, Quebec, Canada

Submitted 2 February 2006; accepted in final form 11 September 2006

Activation of phosphokinase C (PKC) can increase transmitter release at sensory–motor neuron synapses in Aplysia, but the target of PKC phosphorylation has not been determined. One putative target of PKC at synapses is the synaptosomal-associated protein of 25 kDa (SNAP-25), a member of the SNARE protein complex implicated in synaptic vesicle docking and fusion. To determine whether PKC regulated transmitter release through phosphorylation of SNAP-25, we cloned Aplysia SNAP-25 and expressed enhanced green fluorescent protein (EGFP)–coupled SNAP-25 constructs mutated at the PKC phosphorylation site Ser198 in Aplysia sensory neurons. We found several distinct effects of expression of EGFP–SNAP-25 constructs. First, the rates of synaptic depression were slowed when cells contained SNAP-25 with phosphomimetic residues Glu or Asp. Second, PDBu-mediated increases in transmitter release at naïve synapses were blocked in cells expressing nonphosphorylated-state SNAP-25. Finally, expression of EGFP-coupled SNAP-25 but not uncoupled SNAP-25 inhibited 5-HT–mediated reversal of depression and the ability of EGFP-coupled SNAP-25 to inhibit the reversal of depression was affected by changes at Ser198. These results suggest SNAP-25 and phosphorylation of SNAP-25 by PKC can regulate transmitter release at Aplysia sensory–motor neuron synapses by a number of distinct processes.

Address for reprint requests and other correspondence: V. F. Castellucci, Pavillon Paul-G. Desmarais, 2960, Chemin de la tour, CP 6128, Succ. Centre-ville, Montreal, Quebec, Canada H3C 3J7 (E-mail: vincent.castellucci{at}umontreal.ca)




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