Paired Whole Cell Recordings in Organotypic Hippocampal Slices

Chantelle Fourie; Marianna Kiraly; Daniel V. Madison; Johanna M. Montgomery

doi:10.3791/51958

Paired Whole Cell Recordings in Organotypic Hippocampal Slices

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Paired Whole Cell Recordings in Organotypic Hippocampal Slices

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09:23 min

September 28, 2014

DOI: 10.3791/51958-v

Chantelle Fourie¹, Marianna Kiraly², Daniel V. Madison*², Johanna M. Montgomery*¹

¹Department of Physiology and Centre for Brain Research,University of Auckland, ²Department of Molecular and Cellular Physiology,Stanford University

Overview

This article describes a methodology for performing pair recordings from synaptically connected neurons using whole cell patch clamp techniques. The focus is on organotypic hippocampal slice cultures, which allow for detailed electrophysiological characterization of synaptic interactions.

Key Study Components

Area of Science

Neuroscience
Electrophysiology
Synaptic connectivity

Background

Pair recordings enable the study of synaptic circuits between individual neurons.
Organotypic slice cultures maintain appropriate synaptic connections for analysis.
This technique can be applied to various brain regions with connected neurons.
Accurate identification of brain landmarks is crucial for successful recordings.

Purpose of Study

To record electrophysiological activity from minimal synaptic circuits.
To isolate synaptic activity between two neurons.
To study basic properties of neuronal circuitry.

Methods Used

Preparation of organotypic hippocampal slice cultures.
Whole cell patch clamp recordings from connected neurons.
Visual demonstration of recording and dissecting methods.
Preparation of solutions and culture plates prior to experiments.

Main Results

Successful isolation of synaptic activity in minimal circuits.
Demonstration of synaptic connections in cultured hippocampal slices.
Methodology applicable to various brain regions.
Visual aids enhance understanding of the experimental process.

Conclusions

Pair recordings provide insights into synaptic function.
Organotypic cultures are effective for studying neuronal circuitry.
Methodology can be adapted for different areas of the brain.

Frequently Asked Questions

What are pair recordings?

Pair recordings are simultaneous whole cell patch clamp recordings from two synaptically connected neurons.

Why use organotypic slice cultures?

They maintain appropriate synaptic connections and allow for detailed electrophysiological recordings.

What is the significance of studying minimal synaptic circuits?

It helps in understanding the basic properties of neuronal circuitry and synaptic interactions.

How important is visual demonstration in this methodology?

Visual demonstration is crucial for accurately identifying brain landmarks and neurons during the process.

Can this method be applied to other brain regions?

Yes, it can be used for any area of the brain where neighboring neurons are synaptically connected.

What preparations are needed before conducting experiments?

All solutions and culture plates must be prepared prior to making the slices.

Pair recordings are simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling precise electrophysiological and pharmacological characterization of the synapses between individual neurons. Here we describe the detailed methodology and requirements for establishing this technique in organotypic hippocampal slice cultures in any laboratory equipped for electrophysiology.

The overall goal of the following experiment is to be able to record electrophysiological from the minimal synaptic circuit in the brain. This is achieved in recordings made from organotypic slice cultures from rat hippocampus grown in plastic culture inserts. These slices make elaborate but appropriate synaptic connections that can be easily recorded after a period of growth in culture.

Making recordings in these slices allows for the isolation of the synaptic activity of the minimal synaptic circuit that made between only two neurons, and thus allows for the study of the most basic properties of neuronal circuitry. This method can be used to probe the function of synaptic micro circuits in the hippocampus, but it can be used for any area of the brain where neighboring neurons are synaptically connected. Visual demonstration of both the recording and dissecting methods are crucial as many of the steps depend on the accurate identification of brain landmarks and neurons, Both before making slices prepare all solutions and the culture plates.

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