Patch Clamp Recordings on Intact Dorsal Root Ganglia from Adult Rats

Kerui Gong; Peter T. Ohara; Luc Jasmin

doi:10.3791/54287

Patch Clamp Recordings on Intact Dorsal Root Ganglia from Adult Rats

JoVE Journal
Biochemistry

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JoVE Journal Biochemistry

Patch Clamp Recordings on Intact Dorsal Root Ganglia from Adult Rats

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12:00 min

September 29, 2016

DOI: 10.3791/54287-v

Kerui Gong¹, Peter T. Ohara², Luc Jasmin¹

¹Department of Oral and Maxillofacial Surgery,University of California, San Francisco, ²Department of Anatomy,University of California, San Francisco

Overview

This manuscript describes a method for preparing intact dorsal root ganglia (DRG) for patch clamp recordings. This technique preserves the microenvironment of neurons and satellite glial cells, minimizing phenotypic and functional changes.

Key Study Components

Area of Science

Neuroscience
Electrophysiology
Cell Biology

Background

Dorsal root ganglia contain primary sensory neurons and glial cells.
Maintaining intact DRG is crucial for accurate physiological studies.
Patch clamp recordings are essential for studying neuronal activity.
Traditional methods may alter the cellular environment.

Purpose of Study

To introduce a method for preparing intact DRG.
To enable single-cell recordings from DRG while stimulating spinal nerves.
To study the role of primary sensory neurons in pain mechanisms.

Methods Used

Anesthesia of the rat.
Preparation of the lumbar area for access to DRG.
Patch clamp recording techniques.
Stimulation of entering spinal nerves or exiting dorsal roots.

Main Results

The method successfully preserves the microenvironment of DRG.
Single-cell recordings provide insights into neuronal activity.
Maintaining glial cells enhances the physiological relevance of findings.
This approach is beneficial for pain research.

Conclusions

Intact DRG preparations are superior for electrophysiological studies.
The method allows for more accurate modeling of in vivo conditions.
This technique can advance the understanding of sensory neuron functions.

Frequently Asked Questions

What is the significance of using intact DRG?

Using intact DRG maintains the natural cellular environment, leading to more accurate physiological data.

How does this method compare to traditional techniques?

This method avoids the phenotypic changes associated with dissociated neurons, providing more reliable results.

What are the applications of this technique?

It is particularly useful in pain research and studying sensory neuron functions.

What precautions should be taken during preparation?

Ensure proper anesthesia and careful handling of the lumbar area to maintain tissue integrity.

Can this method be applied to other types of neurons?

While this method is optimized for DRG, similar techniques may be adapted for other neuron types.

This manuscript describes how to prepare intact dorsal root ganglia for patch clamp recordings. This preparation maintains the microenvironment for neurons and satellite glial cells, thus avoiding the phenotypic and functional changes seen using dissociated DRG neurons.

The overall goal for this experiment is to introduce methods to prepare the intact dorsal root ganglia and to conduct patch clamp recording using this preparation. This method allows single cell recording from the intact dorsal root ganglia while stimulating the entering spinal nerve or the exiting dorsal root. This approach is especially useful when studying the contribution of primary sensory neurons to pain.

The main advantage of this technique is that it maintains both neuron and adjacent satellite glial cells. Therefore, this preparation is closer to in vivo situation. In this procedure, after the rat has been anesthetized, shave its lumbar area.

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