From Molecular Inventory to Functional Ontology
Proteomic reconstructions suggest that a presynaptic bouton contains roughly 1,400–1,800 protein species, while only about 30–60 are assigned directly to the synaptic vesicle—less than 4% of the inventory. The remaining machinery organizes vesicle positioning, energy supply, cytoskeletal transport, calcium coupling, endocytosis and local signaling. This molecular diversity is also therapeutically important: presynaptic proteins include a substantial fraction of CNS drug targets and many genes associated with neurological and psychiatric disease. We therefore combine systematic perturbation, optical release assays and interpretable AI to connect molecular identity with measurable synaptic function.
Optical measurements reveal the causal chain from calcium entry to neurotransmitter output. The paired recording and Fluo-5F movie at right measures action-potential-evoked calcium influx in individual boutons.
A scalable workflow turns gene perturbations into functional phenotypes. We select presynaptic targets, perform CRISPR–Cas9 knockout in cultured hippocampal networks, and combine automated stimulation with high-speed imaging. Each perturbation is profiled across glutamate and GABA release, paired-pulse behavior, sucrose-defined readily releasable pool, release probability, subcellular localization, network activity and transcriptomics. The resulting atlas distinguishes core release machinery, calcium sensors, vesicle organizers and disease-linked regulators, while revealing unexpected functional convergence among genes.
From Functional Landscapes to Predictive Models
Interpretable AI embeds multi-assay phenotypes into a functional manifold. Proximity in this space reflects shared function rather than sequence alone, exposing modules that govern vesicle priming, calcium coupling, vesicle organization and excitatory-versus-inhibitory transmission. The atlas also makes neuropsychiatric genetics experimentally tractable: autism- and schizophrenia-associated genes can be ranked by the specific release features they disrupt and then returned to focused mechanistic experiments.
This movie shows the Glutamate sensor iGluSnFR4 in response to single electrical stimulation and spontaneous vesicle release.
This movie visualizes fluorescence activity recorded simultaneously across multiple regions of a neuronal network within an approximately 200 µm field of view. Recurrent waves of coordinated activity reveal when local signals become population-level network states.