Imaging mRNA localisation in memory-relevant neurons of the Drosophila brain

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Memories are encoded in Drosophila as dopamine driven changes in the efficacy of synaptic connections between Mushroom Body Kenyon Cells and Mushroom Body Output Neurons (MBONs). Evidence from flies and mammals suggests that some learning-relevant synaptic plasticity requires local translation of new proteins within specific neuronal processes. Neurons therefore have to deliver the relevant mRNAs to these neuronal compartments as a means of spatially and temporally regulating protein synthesis. We used single molecule fluorescent in situ hybridisation (smFISH) and the MS2 system to image mRNA localisation in the adult Drosophila brain, before and after learning. smFISH allowed us to visualise learning induced increases in CaMKII mRNA within the dendritic compartments of specific MBONs following aversive olfactory conditioning. We are now using the MS2 system to live-track mRNA transcripts in memory-relevant neurons of head-fixed flies, after learning. These developments enable the visualisation of molecular correlates of plasticity, and therefore extend the analysis of memory in Drosophila beyond measuring changes in odour-evoked neural activity.