Title: Novel Small-RNA Mediated Gene Regulatory Mechanisms for Long-Term Memory
Description: Memory storage and memory-related synaptic plasticity rely on precise spatiotemporal regulation of gene expression. To explore the role of small RNAs in memory-related synaptic plasticity we carried out massive parallel sequencing to profile the small RNAs of Aplysia. We identified 170 distinct 21-23 nt sized miRNAs, 13 of which were novel and specific to Aplysia. Nine miRNAs were brain-enriched, and several of these were rapidly down-regulated by transient exposure to serotonin, a modulatory neurotransmitter released during learning. Two abundant, and conserved brain-specific miRNAs, miR-124 and miR-22 were exclusively present pre-synaptically in a sensory-motor synapse where they constrain synaptic facilitation through regulation of the transcriptional factor CREB1 and translation factor CPEB respectively. We therefore provide the first evidence that a modulatory neurotransmitter important for learning can regulate the levels of small RNAs and present a novel role for miR-124 and miR-22 in long-term plasticity of synapses in the mature nervous system.

While mining the small RNA libraries for miRNAs, we discovered an unexpected and abundant expression in brain of a 28-nt sized class of piRNAs, which had been thought to be germ-line specific. These piRNAs have unique biogenesis patterns and predominant nuclear localization. Moreover, we find that whereas miRNAs are down-regulated by exposure to serotonin, piRNAs are up-regulated. Importantly, we find that the piwi/piRNA complex facilitates serotonin-dependent methylation of a conserved CpG island in the promoter of CREB2, the major inhibitory constraint of memory in Aplysia, leading to the persistence of long-term synaptic facilitation. Taken together, these findings provide a new serotonin-dependent, bidirectional, small-RNA mediated gene regulatory mechanism during plasticity where miRNAs provide translational control and piRNAs provide long-lasting transcriptional control for the persistence of memory.