Serotonin (5-HT)-induced long-term facilitation (LTF) of the Aplysia sensorimotor synapse depends on enhanced gene expression and protein synthesis, but identification of the genes whose expression and regulation are necessary for LTF remains incomplete. In this study we found that one such gene is synapsin, which encodes a synaptic vesicle-associated protein known to regulate short-term synaptic plasticity. Both synapsin mRNA and protein levels were increased by 5-HT. Upregulation of synapsin protein occurred in presynaptic sensory neurons at neurotransmitter release sites. To investigate the molecular mechanisms underlying synapsin regulation, we cloned the promoter region of Aplysia synapsin, and found that the synapsin promoter contained a cAMP response element (CRE), raising the possibility that the transcriptional activator CREB1 mediates 5-HT-induced regulation of synapsin. Indeed, binding of CREB1 to the synapsin promoter was increased following treatment with 5-HT. Furthermore, increased acetylation of histones H3 and H4 and decreased association of histone deacetylase 5 near the CRE site is consistent with transcriptional activation by CREB1. RNA interference (RNAi) targeting synapsin mRNA blocked the 5-HT-induced increase in synapsin protein levels and LTF during a time window when basal synapsin levels were unaffected by RNAi. These results indicate that the 5-HT-induced regulation of synapsin levels is necessary for LTF and that this regulation is part of the cascade of synaptic events involved in the consolidation of memory.

Cerebral peptide 2 (CP2), a 41 amino acid neuropeptide, was identified because it was transported from the cerebral ganglia of Aplysia to other central ganglia. Immunocytology indicates that CP2 is distributed widely in the CNS and peripheral tissues of Aplysia. Most CP2-immunoreactive neurons were found in the cerebral ganglia and extensively overlap with the distribution of cerebral peptide 1 (CP1). HPLC analyses confirm that individual cerebral neurons synthesize both CP1 and CP2. In other ganglia, CP1 and CP2 are localized predominantly to different neurons. CP2-immunoreactive fibers and varicosities are present in the neuropil of all ganglia but were found surrounding cell bodies and axon hillocks most often in the buccal and abdominal ganglia. Thus, the effects of CP2 on neurons in these ganglia were determined using intracellular recording. In the buccal ganglia, CP2 evokes rhythmic activity in many motor neurons that seems similar to that observed during ingestion; however, only one identified neuron was found to be depolarized directly. By contrast, in the abdominal ganglion, many neurons are depolarized directly by CP2. A number of these have been shown to be part of the circuit that regulates respiratory pumping. Injection of CP2 into freely behaving Aplysia increases the rate of respiratory pumping and causes other changes in behavior. CP2 is stable in hemolymph, which raises the possibility that it may act as a hormone. Thus, CP2 is a bioactive neuropeptide that is present in many neurons and likely functions as a transmitter or a hormone.

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The Aplysia sensorimotor synapse is a key site of plasticity for several simple forms of learning. Plasticity of this synapse has been extensively studied, albeit primarily with individual action potentials elicited at low frequencies. Yet, the mechanosensory neurons fire high-frequency bursts in response to even moderate tactile stimuli delivered to the skin. In the present study, we extend this analysis to show that sensory neurons also fire bursts in the range of 1-60 Hz in response to electrical stimuli similar to those used in behavioral studies of sensitization. Intracellular stimulation of sensory neurons to fire a burst of action potentials at 10 Hz for 1 sec led to significant homosynaptic depression of postsynaptic responses. The depression was transient and fully recovered within 10 min. During the burst, the steady-state depressed phase of the postsynaptic response, which was only 20% of the initial EPSP of the burst, still contributed to firing the motor neuron. To explore the functional contribution of transient homosynaptic depression to the response of the motor neuron, computer simulations of the sensorimotor synapse with and without depression were compared. Depression allowed the motor neuron to produce graded responses over a wide range of presynaptic input strength. In addition, enhancement of synaptic transmission throughout a burst increased motor neuron output substantially more than did preferential enhancement of the initial phase of a burst. Thus, synaptic depression increased the dynamic range of the sensorimotor synapse and can, in principle, have a profound effect on information processing.