Neuroscience

Receptor

Electrophysiology

Calcium imaging

Zebrafish

Functional connectivity

Connectome

Biology

Gene

Biological neural network

Neuroanatomy

Synapse

Artificial intelligence

Postsynaptic potential

Metallurgy

Materials science

Connectomics

Slice preparation

Computer science

Calcium

Biochemistry

Nature Methods

Dense reconstruction of synaptic connectivity requires high-resolution electron microscopy images of entire brains and tools to efficiently trace neuronal wires across the volume. To generate such a resource, we sectioned and imaged a larval zebrafish brain by serial block-face electron microscopy at a voxel size of 14 × 14 × 25 nm 3 . We segmented the resulting dataset with the flood-filling network algorithm, automated the detection of chemical synapses and validated the results by comparisons to transmission electron microscopic images and light-microscopic reconstructions. Neurons and their connections are stored in the form of a queryable and expandable digital address book. We reconstructed a network of 208 neurons involved in visual motion processing, most of them located in the pretectum, which had been functionally characterized in the same specimen by two-photon calcium imaging. Moreover, we mapped all 407 presynaptic and postsynaptic partners of two superficial interneurons in the tectum. The resource developed here serves as a foundation for synaptic-resolution circuit analyses in the zebrafish nervous system.

Automated synapse-level reconstruction of neural circuits in the larval zebrafish brain