Preparation of Rat Sciatic Nerve for <em>Ex Vivo</em> Neurophysiology

Adrien Rapeaux; Omaer Syed; Estelle Cuttaz; Christopher A. R. Chapman; Rylie A. Green; Timothy G. Constandinou

doi:10.3791/63838

Preparación del nervio ciático de rata para neurofisiología ex vivo

JoVE Journal
Neuroscience

This content is Free Access.

JoVE Journal Neuroscience

Preparation of Rat Sciatic Nerve for Ex Vivo Neurophysiology

Preparación del nervio ciático de rata para neurofisiología ex vivo

Full Text

5,248 Views

09:09 min

July 12, 2022

DOI: 10.3791/63838-v

Adrien Rapeaux^1,2,3,4, Omaer Syed⁵, Estelle Cuttaz⁵, Christopher A. R. Chapman⁵, Rylie A. Green⁵, Timothy G. Constandinou^1,2,3,4

¹Department of Electrical and Electronic Engineering,Imperial College London, ²Centre for Bioinspired Technology,Imperial College London, ³Care Research and Technology (CR&T),Imperial College London and the University of Surrey, ⁴Dementia Institute (UK DRI), ⁵Department of Bioengineering,Imperial College London

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article presents a protocol for preparing rat whole sciatic nerve tissue to facilitate ex vivo electrophysiological stimulation and recording. This method is designed to minimize external influences and improve the reliability of neurophysiological data collection.

Key Study Components

Area of Science

Neurophysiology
Electrophysiology
Ex vivo techniques

Background

The sciatic nerve is crucial for studying neurophysiological responses and phenomena.
Prior methods have faced challenges due to variables in living organisms, such as anesthesia depth.
This protocol allows for detailed observation without the confounding factors present in in vivo studies.

Purpose of Study

To enable detailed observations of neurophysiological phenomena such as nerve block carryover.
To establish a reliable ex vivo model that improves measurement precision.
To simplify preparation procedures in comparison to in vivo approaches.

Methods Used

The protocol employs an ex vivo electrophysiological setup using rat sciatic nerve.
Dissection techniques are outlined for carefully extracting the nerve while maintaining moisture.
Critical steps include using chilled modified Krebs-Henseleit buffer during dissection and setting up a dual-chamber nerve bath.
The nerve is secured and subjected to stimulation with electrodes in a controlled environment.

Main Results

This method yields robust and repeatable electrophysiological readings.
Interference from physiological conditions is minimized, resulting in clearer data.
It allows for investigation of specific neurophysiological phenomena in a controlled, dissected environment.

Conclusions

The study demonstrates a successful approach for ex vivo nerve tissue preparation.
This technique enhances the understanding of neurophysiological mechanisms without in vivo variability.
It provides a useful model for exploring nerve function and dysfunction under controlled settings.

Frequently Asked Questions

What are the advantages of using ex vivo techniques?

Ex vivo methods allow for detailed observation of neurological functions in a controlled environment, minimizing confounding factors that affect in vivo studies.

How is the rat sciatic nerve prepared for stimulation?

The sciatic nerve is carefully exposed and detached from surrounding tissues, maintained in a chilled buffer to prevent drying out during the process.

What types of data can be obtained using this protocol?

This protocol provides electrophysiological data, allowing for the observation of nerve responses to specific current injections without physiological interference.

How can this method be adapted for other types of nerves?

The techniques described can be modified to accommodate different types of nerves by adjusting dissection and stimulation protocols based on anatomical variations.

What are the limitations of this approach?

Some potential limitations include the inability to observe whole-body responses and the necessity for precise dissection techniques to prevent nerve damage.

Este protocolo describe la preparación de tejido nervioso ciático entero de rata para la estimulación electrofisiológica ex vivo y el registro en un baño salino perfundido de dos compartimentos, regulado ambientalmente.

Este protocolo permite la observación de muchos fenómenos neurofisiológicos que no se comprenden bien, como el arrastre de bloqueo nervioso después de la inyección de corriente de alta frecuencia / alta amplitud, al tiempo que evita la interferencia de otros procesos en un cuerpo vivo. Esta técnica produce resultados altamente repetibles y robustos y es más práctica que la electrofisiología in vivo, porque las variables complejas como la profundidad de la anestesia no tienen que ser controladas y, por lo tanto, no pueden influir en las mediciones. Después de un procedimiento de eutanasia bajo anestesia, coloque una rata hembra en la mesa de disección, sostenga el tobillo firmemente entre el pulgar, el dedo índice y el dedo medio, y corte el tendón calcánea con tijeras romas rectas de 12 centímetros.

Usando fórceps finos y tijeras, haga incisiones cuidadosas a través de las capas musculares cerca de la mitad de la parte posterior de la pierna hasta que el nervio ciático esté expuesto. Una vez que el nervio sea visible, hidrate la cavidad usando un tampón Krebs-Henseleit modificado con hielo, o MKHB, para evitar que el nervio se seque. Usando hemostáticos, tire de los colgajos de piel de cada lado y mantenga la incisión abierta para una disección más fina.

View the full transcript and gain access to thousands of scientific videos