Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels.
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.
Department of Medicine, Imperial College London, London, W12 0NN, UK; School of Engineering and Materials Science, Queen Mary, University of London, London E1 4NS, UK. Electronic address: p.novak@qmul.ac.uk.
Pavel
P
Novak
Julia
J
Gorelik
Umesh
U
Vivekananda
Andrew I
AI
Shevchuk
Yaroslav S
YS
Ermolyuk
Russell J
RJ
Bailey
Andrew J
AJ
Bushby
Guy W J
GW
Moss
Dmitri A
DA
Rusakov
David
D
Klenerman
Dimitri M
DM
Kullmann
Kirill E
KE
Volynski
Yuri E
YE
Korchev
D000818:Animals; D000831:Animals, Newborn; D055592:Biophysical Phenomena / Q000502:physiology; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D049229:Dendritic Spines / Q000502:physiology / Q000648:ultrastructure; D004558:Electric Stimulation; D004566:Electrodes; D005456:Fluorescent Dyes / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology; D021621:Imaging, Three-Dimensional; D015640:Ion Channel Gating / Q000502:physiology*; D007473:Ion Channels / Q000502:physiology* / Q000648:ultrastructure; D008564:Membrane Potentials / Q000502:physiology; D016252:Microscopy, Scanning Tunneling; D009474:Neurons / Q000166:cytology*; D018408:Patch-Clamp Techniques; D017729:Presynaptic Terminals / Q000502:physiology* / Q000648:ultrastructure; D051381:Rats
D005456:Fluorescent Dyes; D007473:Ion Channels; D002118:Calcium