Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels

Autor: Umesh Vivekananda, Yuri E. Korchev, Kirill E. Volynski, Andrew Shevchuk, Pavel Novák, Andrew J. Bushby, David Klenerman, Guy W. J. Moss, Julia Gorelik, Russell J. Bailey, Yaroslav S. Ermolyuk, Dmitri A. Rusakov, Dimitri M. Kullmann
Přispěvatelé: Biotechnology and Biological Sciences Research Council (BBSRC), Biotechnology and Biological Sciences Research Cou
Jazyk: angličtina
Rok vydání: 2013
Předmět:
Dendritic spine
Patch-Clamp Techniques
1702 Cognitive Sciences
MEMBRANES
Hippocampus
Ion Channels
Membrane Potentials
0302 clinical medicine
Microscopy
Scanning Tunneling
Cells
Cultured
Hippocampal mossy fiber
Membrane potential
Neurons
0303 health sciences
Chemistry
General Neuroscience
MICROSCOPY
CA2+ CHANNELS
3. Good health
Calyx of Held
Life Sciences & Biomedicine
Ion Channel Gating
TRANSMISSION
Neuroscience(all)
Dendritic Spines
Presynaptic Terminals
TERMINALS
Biophysical Phenomena
Ion
03 medical and health sciences
Imaging
Three-Dimensional
Animals
Patch clamp
MODULATION
Electrodes
Ion channel
030304 developmental biology
Fluorescent Dyes
Science & Technology
Neurology & Neurosurgery
TRANSMITTER RELEASE
Neurosciences
NeuroResource
Electric Stimulation
Rats
Electrophysiology
Animals
Newborn
1701 Psychology
Calcium
Neurosciences & Neurology
1109 Neurosciences
Neuroscience
030217 neurology & neurosurgery
Zdroj: Neuron
Scopus-Elsevier
Popis: 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 Ca2+ channels. This semiautomated approach allows direct investigation of the distribution and properties of presynaptic ion channels at small central synapses. Video Abstract
Highlights • Topographic imaging of live synaptic boutons at nanoscale resolution • Cell-attached patch-clamp recordings of ion channels in small central synapses • Whole-cell small presynaptic bouton recordings
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.
Databáze: OpenAIRE
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