Rapid neurotransmission depends on the structural and functional integrity of synaptic connections. How synapses assemble is currently being intensely investigated to help our understanding of neuronal development and synaptic plasticity. Here we focus on the assembly of the presynaptic active zone, which regulates the synaptic vesicle exo/endo-cycle and is characterized by ultrastructural specializations and large scaffold proteins. While genetic and biochemical studies from rodents, Caenorhabditis elegans and Drosophila have started to identify proteins organizing active zone assembly, drawing a coherent picture remains challenging, with genetically established hierarchies and protein-protein interactions still to be placed into spatio-temporal and functional context. Recent advances in light and electron microscopy, together with in vivo imaging of protein traffic, will help to tackle this challenge.