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In Drosophila, Mushroom Bodies (MBs) are the site of CS-US association for olfactory memory.MBs receive olfactory CS inputs from cholinergic projection neurons and dopamine (DA) neurons convey the aversive US.We demonstrate that US information is received solely by the γlobe subset of MB neurons.This is surprising given the known role in memory of signaling within additional MB cell types.Convergent evidence has demonstrated that each of the MB subtypes play important but functionally distinct roles during encoding, consolidation and retrieval.One of the most striking findings in the literature, for example, is that expression of the Rutabaga adenylyl cyclase only in γ neurons can fully restore short-term memory (STM) to rutabaga mutants, whereas expression of NF1 in α/β neurons is sufficient to restore STM to NF 1 mutants.Because these mutants each disrupt only a portion of STM performance, this has been taken as evidence that distinct signaling mechanisms support parallel memory traces within these two sets of neurons.To test this model, we focused on the D l-like Dopamine receptor (DopR), mutations of which completely eliminate performance.We demonstrate that DopR expression in y neurons is sufficient to fully support memory performance.Because of the known role ofMB-MP1 DA neurons in mediating the US, we used GFP-reconstituted across the synapse (GRASP) to visualize synaptic connections between MB-MP1 and γneurons.Our findings support a model in which the CS-US association forms solely in γ neurons.Subsequent memory processing then engages signaling in α/β neurons.