Delayed gramicidin delivery through an intra-pipette capillary facilitates perforated patch recordings

Kosuke Kazashi; Kakeru Miura; Sota Ueda; Kazuhito Utsunomiya; Mari Kiriyama; Motoki Yagisawa; Masayuki Ikeda; Masanobu Kano; Toshihide Tabata

doi:10.1016/j.neures.2024.05.002

Delayed gramicidin delivery through an intra-pipette capillary facilitates perforated patch recordings

Kosuke Kazashi, Kakeru Miura, Sota Ueda, Kazuhito Utsunomiya, Mari Kiriyama, Motoki Yagisawa, Masayuki Ikeda, Masanobu Kano, Toshihide Tabata^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

The gramicidin-perforated patch-clamp technique is indispensable for recording neuronal activities without changing the intracellular Cl⁻ concentration. Conventionally, gramicidin contained in the pipette fluid is delivered to the cell membrane by passive diffusion. Gramicidin deposited on the pipette orifice sometimes hampers giga-seal formation, and perforation progresses only slowly. These problems may be circumvented by delivering a high concentration of gramicidin from an intra-pipette capillary after a giga-seal is formed. We herein describe the detailed protocol of this improved method. This protocol would greatly facilitate the investigation of Cl⁻ gradient-dependent neuronal activities.

Original language	English
Pages (from-to)	45-48
Number of pages	4
Journal	Neuroscience Research
Volume	207
DOIs	https://doi.org/10.1016/j.neures.2024.05.002
State	Published - 2024/10

Keywords

Chloride
Gramicidin
Neuron
Patch-clamp
Perforated-patch
Seal resistance
Series resistance

ASJC Scopus subject areas

General Neuroscience

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10.1016/j.neures.2024.05.002

Cite this

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title = "Delayed gramicidin delivery through an intra-pipette capillary facilitates perforated patch recordings",

abstract = "The gramicidin-perforated patch-clamp technique is indispensable for recording neuronal activities without changing the intracellular Cl− concentration. Conventionally, gramicidin contained in the pipette fluid is delivered to the cell membrane by passive diffusion. Gramicidin deposited on the pipette orifice sometimes hampers giga-seal formation, and perforation progresses only slowly. These problems may be circumvented by delivering a high concentration of gramicidin from an intra-pipette capillary after a giga-seal is formed. We herein describe the detailed protocol of this improved method. This protocol would greatly facilitate the investigation of Cl− gradient-dependent neuronal activities.",

keywords = "Chloride, Gramicidin, Neuron, Patch-clamp, Perforated-patch, Seal resistance, Series resistance",

author = "Kosuke Kazashi and Kakeru Miura and Sota Ueda and Kazuhito Utsunomiya and Mari Kiriyama and Motoki Yagisawa and Masayuki Ikeda and Masanobu Kano and Toshihide Tabata",

note = "Publisher Copyright: {\textcopyright} 2024 The Authors",

year = "2024",

month = oct,

doi = "10.1016/j.neures.2024.05.002",

language = "英語",

volume = "207",

pages = "45--48",

journal = "Neuroscience Research",

issn = "0168-0102",

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T1 - Delayed gramicidin delivery through an intra-pipette capillary facilitates perforated patch recordings

AU - Kazashi, Kosuke

AU - Miura, Kakeru

AU - Ueda, Sota

AU - Utsunomiya, Kazuhito

AU - Kiriyama, Mari

AU - Yagisawa, Motoki

AU - Ikeda, Masayuki

AU - Kano, Masanobu

AU - Tabata, Toshihide

PY - 2024/10

Y1 - 2024/10

N2 - The gramicidin-perforated patch-clamp technique is indispensable for recording neuronal activities without changing the intracellular Cl− concentration. Conventionally, gramicidin contained in the pipette fluid is delivered to the cell membrane by passive diffusion. Gramicidin deposited on the pipette orifice sometimes hampers giga-seal formation, and perforation progresses only slowly. These problems may be circumvented by delivering a high concentration of gramicidin from an intra-pipette capillary after a giga-seal is formed. We herein describe the detailed protocol of this improved method. This protocol would greatly facilitate the investigation of Cl− gradient-dependent neuronal activities.

AB - The gramicidin-perforated patch-clamp technique is indispensable for recording neuronal activities without changing the intracellular Cl− concentration. Conventionally, gramicidin contained in the pipette fluid is delivered to the cell membrane by passive diffusion. Gramicidin deposited on the pipette orifice sometimes hampers giga-seal formation, and perforation progresses only slowly. These problems may be circumvented by delivering a high concentration of gramicidin from an intra-pipette capillary after a giga-seal is formed. We herein describe the detailed protocol of this improved method. This protocol would greatly facilitate the investigation of Cl− gradient-dependent neuronal activities.

KW - Chloride

KW - Gramicidin

KW - Neuron

KW - Patch-clamp

KW - Perforated-patch

KW - Seal resistance

KW - Series resistance

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U2 - 10.1016/j.neures.2024.05.002

DO - 10.1016/j.neures.2024.05.002

M3 - 学術論文

AN - SCOPUS:85192701048

SN - 0168-0102

VL - 207

SP - 45

EP - 48

JO - Neuroscience Research

JF - Neuroscience Research

ER -

Delayed gramicidin delivery through an intra-pipette capillary facilitates perforated patch recordings

Abstract

Keywords

ASJC Scopus subject areas

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