Nanoscale-Targeted Patch-Clamp Recordings of Functional Presynaptic Ion Channels

Summary

Direct electrical access to presynaptic ion channels has hitherto been limited to large specialized terminals such as the calyx of Held or hippocampal mossy fiber bouton. The electrophysiology and ion-channel complement of far more abundant small synaptic terminals (≤1 μm) remain poorly understood. Here we report a method based on superresolution scanning ion conductance imaging of small synapses in culture at approximately 100–150 nm 3D resolution, which allows presynaptic patch-clamp recordings in all four configurations (cell-attached, inside-out, outside-out, and whole-cell). Using this technique, we report presynaptic recordings of K ⁺, Na ⁺, Cl ⁻, and Ca ²⁺ channels. This semiautomated approach allows direct investigation of the distribution and properties of presynaptic ion channels at small central synapses.

Video Abstract

Highlights

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  Topographic imaging of live synaptic boutons at nanoscale resolution

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  Cell-attached patch-clamp recordings of ion channels in small central synapses

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  Whole-cell small presynaptic bouton recordings

Abstract

Novak and colleagues have developed a method for nanoscale-targeted patch-clamp presynaptic recordings in submicrometer central synapses identified using superresolution scanning ion conductance microscopy. This semiautomated approach opens a new window on the physiology of small presynaptic terminals.