Whole-cell Patch-clamp Recordings in Brain Slices

Amir Segev; Francisco Garcia-Oscos; Saïd Kourrich

doi:10.3791/54024

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Method Article

Whole-cell Patch-clamp Recordings in Brain Slices

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DOI:

10.3791/54024

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June 15th, 2016

Amir Segev¹ , Francisco Garcia-Oscos¹ , Saïd Kourrich¹

¹Department of Psychiatry, University of Texas Southwestern Medical Center

Summary

This protocol describes basic procedural steps for performing whole-cell patch-clamp recordings. This technique allows the study of the electrical behavior of neurons, and when performed in brain slices, allows the assessment of various neuronal functions from neurons that are still integrated in relatively well preserved brain circuits.

Abstract

Whole-cell patch-clamp recording is an electrophysiological technique that allows the study of the electrical properties of a substantial part of the neuron. In this configuration, the micropipette is in tight contact with the cell membrane, which prevents current leakage and thereby provides more accurate ionic current measurements than the previously used intracellular sharp electrode recording method. Classically, whole-cell recording can be performed on neurons in various types of preparations, including cell culture models, dissociated neurons, neurons in brain slices, and in intact anesthetized or awake animals. In summary, this technique has immensely contributed to the understanding of passive and active biophysical properties of excitable cells. A major advantage of this technique is that it provides information on how specific manipulations (e.g., pharmacological, experimenter-induced plasticity) may alter specific neuronal functions or channels in real-time. Additionally, significant opening of the plasma membrane allows the internal pipette solution to freely diffuse into the cytoplasm, providing means for introducing drugs, e.g., agonists or antagonists of specific intracellular proteins, and manipulating these targets without altering their functions in neighboring cells. This article will focus on whole-cell recording performed on neurons in brain slices, a preparation that has the advantage of recording neurons in relatively well preserved brain circuits, i.e., in a physiologically relevant context. In particular, when combined with appropriate pharmacology, this technique is a powerful tool allowing identification of specific neuroadaptations that occurred following any type of experiences, such as learning, exposure to drugs of abuse, and stress. In summary, whole-cell patch-clamp recordings in brain slices provide means to measure in ex vivo preparation long-lasting changes in neuronal functions that have developed in intact awake animals.

Introduction

The patch-clamp technique, an electrophysiological technique that has been developed in the late 1970s^1,2, is a primary tool for studying single or multiple ion channel functions in live tissue. Among the different patch configurations that can be achieved, whole-cell patch-clamp recordings allow the study of the electrical behavior of a substantial part of the neuron. Classically, this technique is performed in vitro either on brain slices, freshly dissociated neurons, or on cell culture models³. When performed on neurons in brain slices, this technique presents several advantages. In particular: (i) neurons are recorded in relatively p....