Synaptic connectivity mapping among thousands of neurons via parallelized intracellular recording with a microhole electrode array

Abstract

The massive parallelization of neuronal intracellular recording, which enables the measurement of synaptic signals across a neuronal network, and thus the mapping and characterization of synaptic connections, is an open challenge, with the state of the art being limited to the mapping of about 300 synaptic connections. Here we report a 4,096 platinum/platinum-black microhole electrode array fabricated on a complementary metal-oxide semiconductor chip for parallel intracellular recording and thus for synaptic-connectivity mapping. The microhole–neuron interface, together with current-clamp electronics in the underlying semiconductor chip, allowed a 90% average intracellular coupling rate in rat neuronal cultures, generating network-wide intracellular-recording data with abundant synaptic signals. From these data, we extracted more than 70,000 plausible synaptic connections among more than 2,000 neurons and catalogued them into electrical synaptic connections and into inhibitory, weak/uneventful excitatory and strong/eventful excitatory chemical synaptic connections, with an estimated overall error rate of about 5%. This scale of synaptic-connectivity mapping and the ability to characterize synaptic connections is a step towards the functional connectivity mapping of large-scale neuronal networks. The parallel intracellular recording of neurons via an array of 4,096 microhole electrodes on a semiconductor chip enables the large-scale mapping of synaptic connections, as shown with rat neuronal cultures.