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

Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels
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 Ca2+ channels. This semiautomated approach allows direct investigation of the distribution and properties of presynaptic ion channels at small central synapses.
0896-6273
1067-1077
Novak, Pavel
14838d74-6ee4-46b7-acf5-23bb58b2b68e
Gorelik, Julia
f3758a00-fd18-4b15-9ac8-75756a3989a0
Vivekananda, Umesh
eeaae042-c083-48e9-a277-ba9066a0c996
Shevchuk, Andrew I.
8388aa54-775a-47cf-b18b-a8ecf75ae996
Ermolyuk, Yaroslav S.
2bb14fb0-5342-408a-8dad-a89ceab8c7c1
Bailey, Russell J.
73907b06-34b2-4e69-bd96-682541497196
Bushby, Andrew J.
bbff3706-c70c-403c-b9dc-cee1a7a73eff
Moss, Guy W.J.
46f1af0e-69d7-4053-8fc3-3381492c932c
Rusakov, Dmitri A.
f42a893e-828e-4a39-a668-aec4b8a20f16
Klenerman, David
c9b05f20-3859-4f59-9ecb-3320a5e4352c
Kullmann, Dimitri M.
7425b0ef-50ce-4696-8275-058056c5f7dc
Volynski, Kirill E.
a1c7d951-a201-4580-b887-858c11aa45e4
Korchev, Yuri E.
e53ab2e4-cef8-4439-9cef-b1ca04c3b489
Novak, Pavel
14838d74-6ee4-46b7-acf5-23bb58b2b68e
Gorelik, Julia
f3758a00-fd18-4b15-9ac8-75756a3989a0
Vivekananda, Umesh
eeaae042-c083-48e9-a277-ba9066a0c996
Shevchuk, Andrew I.
8388aa54-775a-47cf-b18b-a8ecf75ae996
Ermolyuk, Yaroslav S.
2bb14fb0-5342-408a-8dad-a89ceab8c7c1
Bailey, Russell J.
73907b06-34b2-4e69-bd96-682541497196
Bushby, Andrew J.
bbff3706-c70c-403c-b9dc-cee1a7a73eff
Moss, Guy W.J.
46f1af0e-69d7-4053-8fc3-3381492c932c
Rusakov, Dmitri A.
f42a893e-828e-4a39-a668-aec4b8a20f16
Klenerman, David
c9b05f20-3859-4f59-9ecb-3320a5e4352c
Kullmann, Dimitri M.
7425b0ef-50ce-4696-8275-058056c5f7dc
Volynski, Kirill E.
a1c7d951-a201-4580-b887-858c11aa45e4
Korchev, Yuri E.
e53ab2e4-cef8-4439-9cef-b1ca04c3b489

Novak, Pavel, Gorelik, Julia, Vivekananda, Umesh, Shevchuk, Andrew I., Ermolyuk, Yaroslav S., Bailey, Russell J., Bushby, Andrew J., Moss, Guy W.J., Rusakov, Dmitri A., Klenerman, David, Kullmann, Dimitri M., Volynski, Kirill E. and Korchev, Yuri E. (2013) Nanoscale-targeted patch-clamp recordings of functional presynaptic ion channels. Neuron, 79 (6), 1067-1077. (doi:10.1016/j.neuron.2013.07.012).

Record type: Article

Abstract

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.

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1-s2.0-S0896627313006120-main.pdf__tid=fcef532e-44ad-11e4-857d-00000aacb361&acdnat=1411647659_65241f329d05fb9d8123e5616798ace1 - Version of Record
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Published date: 18 September 2013
Organisations: Optoelectronics Research Centre

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Local EPrints ID: 369405
URI: http://eprints.soton.ac.uk/id/eprint/369405
ISSN: 0896-6273
PURE UUID: 0647ad4b-7752-4773-b43e-933276e734b9

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Date deposited: 25 Sep 2014 12:18
Last modified: 14 Mar 2024 18:02

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Contributors

Author: Pavel Novak
Author: Julia Gorelik
Author: Umesh Vivekananda
Author: Andrew I. Shevchuk
Author: Yaroslav S. Ermolyuk
Author: Russell J. Bailey
Author: Andrew J. Bushby
Author: Guy W.J. Moss
Author: Dmitri A. Rusakov
Author: David Klenerman
Author: Dimitri M. Kullmann
Author: Kirill E. Volynski
Author: Yuri E. Korchev

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