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During the previous funding period, we established proximity biotinylation approaches to map protein organization within presynaptic subcompartments both in vivo and in vitro. These studies enabled the spatial mapping of signaling pathways that may regulate presynaptic functions. In the upcoming funding period, we will investigate how key signaling proteins shape synaptic transmission properties, thereby contributing to heterogeneity among synapses of the same neuron and diversity across synapses of distinct neuronal subtypes.